Pseudomonas

Jpn J Exp Med, 1977 Jun, 47(3), 203 - 8
Serological typing of Pseudomonas aerugionosa--grouping of serotypes; Terada Y et al.; Cross absorption and agar gel diffusion tests were performed using Homma and Liu standard strains and their antisera to study the antigenic relationships among the strains showing strong cross-agglutination . The results suggested that Homma 0:3 strain was able to produce a sufficiently high-titered antiserum against Liu 0:7 and 0:8, and that Liu 0:13 was identical to Homma 0:12 . In the antisera against Liu 0:13 and 0:14 there existed a strong antibody common to both antigens as well as an antibody specific to each antigen . Furthermore, the presence of strong common antigens was demonstrated among Homma 0:2, 0:7, 0:13 and 0:16, and between Homma 0:15 and 0:17.

Can J Microbiol, 1977 Jun, 23(6), 672 - 9
Enzymatic hydrolysis of agar: purification and characterization of beta-neoagarotetraose hydrolase from Pseudomonas atlantica; Groleau D et al.; Agarose is degraded by a beta-agarase from Pseudomonas atlantica to neoagarooligosaccharides of degree of polymerization (DP), 4, 6, 8, and 10 . A beta-neoagarotetraose hydrolase cleaves the central beta-linkage in neoagarotetraose and the beta-linkage near the nonreducing end in neoagarohexaose and -octaose to yield neoagarobiose . The beta-neoagarotetraose hydrolase was localized on or outside the cytoplasmic membrane, in the cell wall region . The enzyme was activated by NaCl, KCl, CaCl2, MnCl2, and MgSO4, has a Km of 3.4 X 10(-3) M for neoagarotetraose, was free from beta-agarase and alpha-neoagarobiose hydrolase activity, and showed no transglycosidic activity.

Biochim Biophys Acta, 1977 May 27, 492(1), 156 - 62
The quatenary structure of Pseudomonas cytochrome oxidase studied by electron microscopy; Saraste M et al.; Pseudomonas cytochrome oxidase (EC 1.9.3.2) was studied by negative staining in the electron microscope . The best resolution was obtained with uranyl oxalate (pH 6.0) as negative stain . Electron micrographs confirm the idea of the dimeric structure of the enzyme . A rough model of cytochrome oxidase was constructed based on different projections of the molecule seen in the electron micrographs . In this model the subunits are identical and sterically equivalent.

Biochemistry, 1977 May 17, 16(10), 2181 - 8
Purification and properties of gamma-butyrobetaine hydroxylase from Pseudomonas sp AK 1; Lindstedt G et al.; gamma-Butyrobetaine hydroxylase (4-trimethylaminobutyrate, 2-oxoglutarate:oxygen oxidoreductase (3-hydroxylating), EC 1.14.11.1) has been isolated from Pseudomonas sp AK 1 by ion-exchange, adsorption, and molecular-sieving chromatography . The preparation was homogeneous as judged from electrophoresis in agarose and polyacrylamide gels, isoelectric focusing, and equilibrium sedimentation . The molecular mass was 95 kdaltons as determined by sedimentation equilibrium centrifugation . From electrophoresis in polyacrylamide gel the molecular mass was estimated to 92 kdaltons, from gel filtration through columns of Sephadex G-200 to 86 kdaltons, and from gel filtration through thin layers of Sephadex G-150 and G-200 to 82 kdaltons . Calculation of molecular mass from Stokes radius, sedimentation coefficient, and partial specific volume gave a value of 96 kdaltons, and from the sedimentation coefficient, 93 kdaltons . Gel filtration through Sephadex G-200 in 6 M guanidinium chloride and electrophoresis in polyacrylamide gel containing 3.5 mM sodium dodecyl sulfate resulted in single bands with mobilities corresponding to molecular masses of 39 and 37 kdaltons, respectively, indicating that the enzyme is composed of two polypeptides chains with similar size . NH2-terminal amino acid sequencing in three cycles resulted in two amino acids in each cycle (Ala + Asn, Ala + Ile, Ala + Ile) . The Stokes radius was 3.8 nm, corresponding to a diffusion coefficient of 5.7 X 10(-7) cm2/s . A sedimentation coefficient of 5.8 X 10(-13) s and a frictional ratio of 1.26 was found . The partial specific volume was 0.729 mL/g at 20 degrees C as calculated from amino acid analysis . The isoelectric point was 5.1, as determined by isoelectric focusing analysis . The light absorption in the ultraviolet and visible regions was that of a protein without light-absorbing prosthetic groups . The absorption coefficient at 280 nm of a 1.0% solution at pH 6.5 was 12.6 . Amino acid analysis by ion-exchange chromatography showed a half-cystine content of 19 mol per 95 kg of protein (23 residues/1000) . Thirteen sulfhydryl groups were found by colorimetric analysis before as well as after reduction with NaBH4, indicating absence of disulfide bonds . Less than 0.1 mol of iron was found per mol of enzyme.

Acta Chir Belg, 1977 May-Jun, 76(3), 317 - 22
{Repair problems of infected iatrogenic lesions of the femoral trifurcation . Review of 7 cases (author's transl)}; Guidicelli H et al.; The authors report on seven cases of infected iatrogenic lesions of the femoral trifurcation . One case was a false aneurism consecutive to a femoral vein catheterization . In 5 cases an arterial lesion followed angiography according to Seldinger . In the last case aetiology was undetermined . Local symptomatology was always associated to systemic infection . The causative germ was a pseudomonas in 3 cases an a staphylococ in 4 others . Operation was performed in emergency in all cases because of a fissure or a rupture of the artery . The authors performed: one simple arterial suture, one resection with plastic graft, one resection with venous graft and 4 resections with double vein graft . This last procedure appears useful for lesions situated at the femoral division.

Mikrobiologiia, 1977 May-Jun, 46(3), 414 - 7
{Pure culture of bacteria using chromates and bichromates as hydrogen acceptors during development under anaerobic conditions}; Romanenko VI et al.; A pure bacterial culture utilizing chromates and bichromates as oxygen donors during growth on organic substances in anaerobic conditions was isolated from active ooze of sewage and industrial wastes containing a weak solution of chromates . A medium for growth of the bacterium was selected, and its morphological and cultural properties were studied . Chromates and bichromates are reduced only in anaerobic conditions by the bacterium or active ooze containing it, the hexavalent chromium becoming trivalent . The culture does not belong to any known species and has been therefore classed as Pseudomonas dechromaticans.

J Gen Virol, 1977 May, 35(2), 353 - 9
Ultrastructure of bacteriophage phi 6: arrangement of the double-stranded RNA and envelope; Gonzalez CF et al.; Pseudomonas phaseolicola cells synchronously infected with phi 6 were fixed and embedded by several procedures . The diameter of phi 6 measured 75 nm and its nucleocapsid 60 nm . Virus nucleocapsids were icosahedral and surrounded by a double membrane . In planar sections the nucleic acid appeared as a hexagonal ring and in cross section as two angular structures.

Exp Hematol, 1977 May, 5(3), 186 - 90
Mobilization of hematopoietic stem cells (CFU-C) into the peripheral blood of man by endotoxin; Cline MJ et al.; Pseudomonas endotoxin was administered to three normal subjects in order to assess possible effects on stem cell circulation . Increased numbers of granulopoietic stem cells (CFU-C) transiently appeared in the peripheral blood, reaching a maximum after the time of maximal leukopenia . There was a mean 4-fold increase in CFU-C per ml of blood following endotoxin . A high proportion of these CFU-C did not require an added source of colony-stimulating activity for growth in agar.

Can J Microbiol, 1977 May, 23(5), 483 - 90
Further studies on the formation of choline sulfate by bacteria; Fitzgerald JW et al.; Cell extracts of Pseudomonas C12B synthesized choline sulfate (COS) from SO42-, choline chloride, and ATP . However, most of the COS-forming activity was found in culture medium supernatants of this bacterium, and that which remained with the cells was cell wall-associated . Enzyme release was independent of the carbon and (or) sulfur source used for growth and was not suppressed by increasing the divalent cation concentration of the medium . The COS-synthesizing system was inhbited in vitro by L-cysteine (greater than or equal to 10(-3) mM), SO42- (greater than 0.1 mM), and choline chloride (greater than 0.1 M) . L-Cysteine (0.1-5.0 mM) did not repress the synthesis of enzymes present in the system . COS formation from SO42- in vitro was increased 2.8-fold by 10 mM adenosine 5'-phosphosulfate (APS) and 5-fold by 1 mM 3'-phosphoadenosine,5'-phosphosulfate (PAPS) during a 4-h incubation period . APS (10 mM) also inhibited the incorporation of 35SO42- into COS . Culture supernatants incubated with Na235SO4 produced two 35S-labelled metabolites having electrophoretic mobilities similar to those exhibited by authentic APS and PAPS . The synthesis of these metabolites was also inhibited in vitro by unlabelled APS and by L-cysteine.

Am Rev Respir Dis, 1977 May, 115(5), 861 - 5
Lung abscess due to Pseudomonas cepacia; Poe RH et al.; A diabetic patient with pneumonia of unspecified origin developed a lung abscess after therapy with ultrasonic nebulization . The etiologic organism was identified as Pseudomonas cepacia . Investigation determined the source of the organism to be the reservoir of the ultrasonic nebulizer, to which the patient was directly exposed through removal of the bottom of the disposable medication cup . When this organism is isolated a nosocomial source of infection should be suspected.

J Infect Dis, 1977 May, 135(5), 729 - 35
Pseudomonas species bacteremia caused by contaminated normal human serum albumin; Steere AC et al.; In May and June 1973, 11 patients on the surgical service at the University of Maryland Hospital had bacteremia caused by Pseudomonas species . Seven of the isolates recovered from blood cultures had the same antibiogram (sensitive only to chloramphenicol and tetracycline) . Ten of the 11 patients were given 25% normal serum albumin (human) shortly before the onset of symptoms . In contrast, only two of seven patients with bacteremia due to Psuedomonas aeruginosa in May and June (P =0.013) and only nine of 20 patients located in surgical special care units during these months (P =0.014) were given this product . When cultured, the albumin in one of 54 previously unopened vials from the implicated lot yielded Pseudomonas cepacia sensitive only to chloramphenicol, tetracycline, and nalidixic acid . Subsequent investigation showed that five more patients in four other hospitals had symptoms of bacteremia shortly after the infusion of different lots of albumin from the same manufacturer, and in four cases P . cepacia was cultured from the suspect albumin . Since sterility testing by manufacturers may not detect low-frequency contamination, surveillance of nosocomial infections, investigation of unusual disease clusters, and prompt reporting of suspect reactions are essential in the control of such outbreaks.

Biochim Biophys Acta, 1977 Apr 27, 497(2), 586 - 97
Oxidation of C1-compounds in Pseudomonas C; Ben-Bassat A et al.; Extracts of Pseudomonas C grown on methanol as a sole carbon and energy source contain a methanol dehydrogenase activity which can be coupled to phenazine methosulfate . This enzyme catalyzes two reactions namely the conversion of methanol to formaldehyde (phenazine methosulfate coupled) and the oxidation of formaldehyde to formate (2,6-dichloroindophenol-coupled) . Activities of glutathione-dependent formaldehyde dehydrogenase (NAD+) and formate dehydrogenase (NAD+) were also detected in the extracts . The addition of D-ribulose 5-phosphate to the reaction mixtures caused a marked increase in the formaldehyde-dependent reduction of NAD+ or NADP+ . In addition, the oxidation of {14C}formaldehyde to CO2, by extracts of Pseudomonas C, increased when D-ribulose 5-phosphate was present in the assay mixtures . The amount of radioactivity found in CO2, was 6;8-times higher when extracts of methanol-grown Pseudomonas C were incubated for a short period of time with {1-14C}glucose 6-phosphate than with {U-14C}glucose 6-phosphate . These data, and the presence of high specific activities of hexulose phosphate synthase, phosphoglucoisomerase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase indicate that in methanol-grown Pseudomonas C, formaldehyde carbon is oxidized to CO2 both via a cyclic pathway which includes the enzymes mentioned and via formate as an oxidation intermediate, with the former predominant.

J Biol Chem, 1977 Apr 25, 252(8), 2648 - 56
Crystalline hemoprotein from Pseudomonas that catalyzes oxidation of side chain of tryptophan and other indole derivatives; Takai K et al.; A new enzyme which catalyzes the oxidation of the side chain of tryptophan and other indole derivatives, has been purified to apparent homogeneity from Pseudomonas and crystallized . The overall purification was about 25-fold with a yield of 4.5% . The purified enzyme was apparently homogeneous as judged by polyacrylamide gel electrophoresis . The molecular weight estimated by gel filtration was approximately 280,000 and sedimentation coefficient (S20,w) was 11 by sucrose density gradient ultracentrifugation . The absorption spectra indicated that the enzyme was a hemoprotein . The purified enzyme was shown to catalyze the reaction in which 1 mol each of NH3 and CO2 was formed at the expense of 1 mol each of L-tryptophan and molecular oxygen . Neither peroxidase nor catalase activity was detected in the purified enzyme and no formation of H2O2 was observed during the enzyme reaction . The product(s) of the reaction was unstable but was converted to and was identified as its stable quinoxaline derivative, 2-(3-indolyl)quinoxaline, in the presence of o-phenylenediamine . These results indicate that the product of the reaction was 3-indolylglycoaldehyde or 3-indolylglyoxal . A variety of other indole derivatives such as D-tryptophan, 5-hydroxyl-L-tryptophan, tryptamine, serotonin, melatonin, N-acetyl-L-tryptophan, N-acetyl-L-tryptophanamide, 3-indoleacetamide, 3-indolelactic acid, 3-indolepropionic acid, 3-indoleethanol, and skatole were also substrates.

J Biol Chem, 1977 Apr 25, 252(8), 2640 - 7
Isolation, crystallization, and properties of indolyl-3-alkane alpha-hydroxylase . A novel tryptophan-metabolizing enzyme; Roberts J et al.; Indolyl-3-alkane alpha-hydroxylase, a novel tryptophan-metabolizing enzyme, was prepared in crystalline form from soil isolate organism Pseudomonas XA . Emission spectroscopy and atomic absorption analyses of purified enzyme revealed the presence of iron (0.8 mol/mol of protein), and a number of observations supported the presence of heme prosthetic group (1.1 mol/mol of protein) . The S20,w value of indolyl-3-alkane alpha-hydroxylase is 10.2 S, and the molecular weight by sedimentation equilibrium ultracentrifugation is 250,000 . The E1%280 of the enzyme is 21, and the isoelectric point by isoelectric focusing on ampholine polyacrylamide gel plates is 4.8 . The enzyme catalyzes hydroxylation on the side chain of a variety of 3-substituted indole compounds, including certain tryptophan-containing oligopeptides . The reaction product from tryptamine was identified by proton nuclear magnetic resonance and gas chromatography/mass spectroscopy analyses . While the indole ring remained intact, hydroxylation occurred at the side chain carbon adjacent to the ring . Nuclear magnetic resonance studies indicated that hydroxylation always took place at the same position when the substrate was tryptophan methyl ester, tryptophol, indole-3-propionate, or indole-3-butyrate . No other chemical change occurred when these substrates were incubated with the enzyme . The Km value of indolyl-3-alkane alpha-hydroxylase for L-tryptophan is 2.4 X 10(-6) M, at pH 7.2 . The enzyme is inhibited by potassium cyanide (0.1 mM) or hydroxylamine (1mM), but not by NaBH4 (25 mM), aminooxyacetic acid (7mM), quinacrine (1 mM), chlortetracycline (1 mM), p-mercuribenzoate (0.1 mM), or ethylenediaminetetraacetate (1 mM) . The plasma half-life (t1/2) of indolyl-3-alkane alpha-hydroxylase in tumor-bearing mice is approximately 25 h.

J Biol Chem, 1977 Apr 10, 252(7), 2390 - 5
Preparation and properties of immobilized rubredoxin; May W et al.; Rubredoxin, one of the three protein components of the epoxidation/hydroxylation system of Pseudomonas oleovorans was immobilized by attachment to CNBr-activated agarose (Sepharose 4B) . Since this represents the first reported example of the preparation of a water-insoluble derivative of an enzyme of this type, the electron transfer and physical properties of the conjugate were examined in order to allow comparison with those of the soluble enzyme . Immobilized rubredoxin exhibits all of the major spectral properties of the soluble enzyme above 300 nm, but some distortion in the 280 nm abosrbance band was observed . The immobilized enzyme accepts electrons from dithionite or form NADPH in the presence of spinach ferredoxin-NADP reductase, and upon reduction the visible absorbance is bleached . Immobilized rubredoxin mediates the reduction of cytochrome c in the presence of NADPH and spinach reductase, although it is less efficient in this role than soluble rubredoxin . The oxidation-reduction potential of immobilized rubredoxin was determined and found to be similar to that of the soluble enzyme . In the presence of 2.5 m guanidine HCL, the immobilized enzyme is considerably more stable than soluble rubredoxin toward denaturation . After anaerobic reduction, iron was readily removed from immobilized rubredoxin by washing in 0.5 m Tris base, PH 9.5 containing 0.07 M mercaptoethanol, and the resulting immobilized apoenzyme could then be reconstituted to give back a conjugate with the original iron content, as judged from its absorbance at 497 NM . Reptition of the entire reduction-dissociation-reconstitution cycle gave the same results as were obtained after the initial reconstitution.

Sem Hop, 1977 Apr 9-16, 53(14-15), 827 - 31
{Intestinal bacterial flora in patients with colopathies . Study of samples obtained by colonoscopy}; Boisson J et al.; This enquiry, the limits of which are easy to determine, permits one to note that in patients with chronic hemorrhagic colonic disease samples of digestive juice, carried, out at the level of the colonic mucosa, permit isolation of a large number of pseudomonadaceae, these bacteria are extremely various . A single patient may harbour 10 to 12 species . These bacteria, usually harmless, belong to the natural microbiocenoses, but they may under certain circumstances, increase in number and, according to Fabiani, induce severe symptoms or even be fatal . The mass of bacteria isolated and identified did not permit a study of bacterial sensitivity . The most seriously affected subjects clinically, usually had various bacteria which excludes the accusation of one species and eliminates any idea of bacterial specificity in the constitution of the syndromes observed . The disturbance seems to be a consequence of these chronic diseases and not the cause . In fact, no pathogenic germ was isolated, which one might consider to be responsible for the disease . Finally, it is wise to conclude that patients with irritable colon syndromes who formed part of this investigation, simply presented some imbalance in the bacterial flora which was sometimes very marked with a predominance of proteolytic Pseudomonas.

Biochem J, 1977 Apr 1, 163(1), 173 - 5
Presence of 3-O-methyl-l-xylose in the lipopolysaccharide of Pseudomonas maltophilia N.C.T.C . 10257; Brown F et al.; 3-O-Methyl-L-xylose was isolated from whole cells of Pseudomonas maltophilia N.C.T.C . 10257 . The sugar is a component of lipopolysaccharide from which a polysaccharide also containing L-rhamnose and L-xylose was released by mild acid hydrolysis . 3-O-Methyl-L-xylose was absent from five other strains of Ps . maltophilia and one strain of Pseudomonas geniculata.

Appl Environ Microbiol, 1977 Apr, 33(4), 881 - 4
N-Alkane oxidation enzymes of a pseudomonad; Parekh VR et al.; A nicotinamide adenine dinucleotide (NAD)-dependent n-alkane dehydrogenase and an NAD phosphate (reduced form)-dependent alkane hydroxylase have been purified from cell-free extracts of Pseudomonas sp . strain 196Aa grown anaerobically on n-alkane . The n-alkane dehydrogenase (fraction R-3), obtained as a single peak from Bio-Gel P-60, showed an overall 135-fold purification and was demonstrated by infrared spectroscopy and gas chromatography to convert n-decane to 1-decene . The alkene hydroxylase activity in the S-3 fraction, purified 167 times from diethylaminoethyl-cellulose, was shown by the same methodology to convert decene to decanol . Commercial ferredoxin has been shown to increase the alkane dehydrogenase activity . An NAD-, flavine adenine dinucleotide-, and iron-dependent alcohol dehydrogenase was demonstrated in the R-3 fraction . A mechanism for the anaerobic conversion of n-alkane to fatty acid has been proposed.

Can J Microbiol, 1977 Apr, 23(4), 476 - 8
Effect of strain variation and growth phase of culture on dry weight and hexosamine content of cell wall layers of a marine pseudomonad; Sussman R et al.; Two variants of marine pseudomonad B-16 (ATCC 19855) differing in that one, variant 3, formed opaque colonies and the other, variant 7, formed translucent colonies were examined to determine if the variants differed in the amount and hexosamine content of their three outer cell wall layers . In both variants, the three outer layers of the cell wall, the loosely bound outer layer, the outer double-track layer, and the underlying (periplasmic space) layer contributed less to the dry weight of the cells when the cells were harvested in the stationary than in the logarithmic phase of growth . The hexosamine content of the layers of variant 3 increased dramatically as the cells went from the logarithmic to the stationary phase . The hexosamine content of the layers of variant 7 changed little by comparison . Thus cells of the variant which forms opaque colonies enrich the outer layers of their cell wall with hexosamine when grown to stationary phase.

J Bacteriol, 1977 Apr, 130(1), 37 - 47
Third system for neutral amino acid transport in a marine pseudomonad; Pearce SM et al.; Uptake of leucine by the marine pseudomonad B-16 is an energy-dependent, concentrative process . Respiratory inhibitors, uncouplers, and sulfhydryl reagents block transport . The uptake of leucine is Na+ dependent, although the relationship between the rate of leucine uptake and Na+ concentration depends, to some extent, on the ionic strength of the suspending assay medium and the manner in which cells are washed prior to assay . Leucine transport can be separated into at least two systems: a low-affinity system with an apparent Km of 1.3 X 10(-5) M, and a high-affinity system with an apparent Km of 1.9 X 10(-7) M . The high-affinity system shows a specificity unusual for bacterial systems in that both aromatic and aliphatic amino acids inhibit leucine transport, provided that they have hydrophobic side chains of a length greater than that of two carbon atoms . The system exhibits strict stereospecificity for the L form . Phenylalanine inhibition was investigated in more detail . The Ki for inhibition of leucine transport by phenylalanine is about 1.4 X 10(-7) M . Phenylalanine itself is transported by an energy-dependent process whose specificity is the same as the high-affinity leucine transport system, as is expected if both amino acids share the same transport system . Studies with protoplasts indicate that a periplasmic binding protein is not an essential part of this transport system . Fein and MacLeod (J . Bacteriol . 124:1177-1190, 1975) reported two neutral amino acid transport systems in strain B-16: the DAG system, serving glycine, D-alanine, D-serine, and alpha-aminoisobutyric acid; and the LIV system, serving L-leucine, L-isoleucine, L-valine, and L-alanine . The high-affinity system reported here is a third neutral amino acid transport system in this marine pseudomonad . We propose the name "LIV-II" system.

J Biochem (Tokyo), 1977 Apr, 81(4), 851 - 8
Purification and some properties of cyclohexylamine oxidase from a Pseudomonas sp; Tokieda T et al.; Cyclohexylamine oxidase was purified 90-fold from cell-free extracts of Pseudomonas sp . capable of assimilating sodium cyclamate . The purified enzyme was homogeneous in disc electrophoresis, and the molecular weight was found to be approximately 80,000 by gel filtration . The enzyme catalyzed the following reaction: cyclohexylamine+O2+H2O leads to cyclohexanone+NH3+H2O2 . The enzyme thus can be classified as an amine oxidase; it utilized oxygen as the ultimate electron acceptor . The pH optimum of the reaction was 6.8 and the apparent Km value for cyclohexylamine was 2.5 X 10(-4) M . The enzyme was highly specific for the deamination of alicyclic primary amines such as cyclohexylamine, but was found to be inactive toward ordinary amines used as substrates for amine oxidases . The enzyme solution was yellow in color and showed a typical flavoprotein spectrum; the addition of cyclohexylamine under anaerobic conditions caused reduction of the flavin in the native enzyme . The flavin of the prosthetic group was identified as FAD by thin layer chromatography . The participation of sulfhydryl groups in the enzymic action was also suggested by the observation that the enzyme activity was inhibited in the presence of PCMB and could be recovered by the addition of glutathione.

J Bacteriol, 1977 Apr, 130(1), 131 - 5
Formation and dissimilation of oxalacetate and pyruvate Pseudomonas citronellolis grown on noncarbohydrate substrates; O'Brien RW et al.; Metabolism of lactate as a carbon source by Pseudomonas citronellolis occurred via a nicotinamide adenine dinucleotide (NAD)-independent L-lactate dehydrogenase, which was present in cells grown on DL-lactate but was not present in cells grown on acetate, aspartate, citrate, glucose, glutamate, or malate . The cells also possessed a constitutive, NAD-independent malate dehydrogenase instead of the conventional NAD-dependent malate dehydrogenase instead of the conventional NAD-dependent enzyme in the tricarboxylic acid cycle . Both enzymes were particulate and used dichlorophenolindo-phenol or oxygen as an electron acceptor . In acetate-grown cells, the activity of pyruvate dehydrogenase and NAD phosphate-linked malate enzyme decreased, cells grown on glucose or lactate . This was consistent with the need to maintain a supply of oxalacetate for metabolism of acetate via the tricarboxylic acid cycle . Changes in enzyme activities suggest that gluconeogenesis from noncarbohydrate carbon sources occurs via the malate enzyme (when oxalacetate decarboxylase is inhibited) or a combination of the NAD-independent malate dehydrogenase and oxalacetate decarboxylase.

Eur J Biochem, 1977 Apr 1, 74(2), 319 - 27
Anabolic ornithine carbamolytransferase of Pseudomonas . The bases of its functional specialization; Stalon V et al.; The anabolic ornithine carbamoyltransferase of Pseudomonas appears to be extremely specialized . Unlike the other carbamoyltransferases studied, this enzyme catalyzes the phosphorolytic cleavage of citrulline with a very poor efficiency . The main goal of this paper is to understand what, in the catalytic process, causes this directed functional specialization . On the basis of kinetic data and thermodynamic properties of the reaction, it appears that the reaction mechanism is the same as for ornithine carbamoyltransferases from other sources, that is, of the sequential ordered type, where carbamoylphosphate is the first substrate to be bound and phosphate the last product to be released . In addition to this, and here lies the difference with other ornithine carbamoyltransferases, the anabolic transferase of Pseudomonas forms a binary dead-end complex with citrulline, leading to inefficient binding of phosphate and citrulline to the enzyme . Therefore the phosphorolytic cleavage of citrulline is equally inefficient . It should be mentioned that the affinity of the enzyme for citrulline at its catalytic site is low as compared to other transferases.

Mikrobiologiia, 1977 Mar-Apr, 46(2), 270 - 2
{Nitrogen fixation in combined cultures of Lipomyces and bacteria}; Bab'eva IP et al.; Nitrogen fixation by pure cultures of soil yeasts belonging to the genus Lipomyces Lodder et Kreger van Rij was not found by the acetylene method on a medium containing microelements and yeast autolysate . In a binary culture of L . lipofer 133 and Pseudomonas sp . 5, a nitrogen fixing bacterium, the content of bound nitrogen was 3.6 times higher in aerobic conditions and 15 times higher in anaerobic conditions than its accumulation by individual bacterial culture.

J Biochem (Tokyo), 1977 Mar, 81(3), 555 - 62
Fine structure of SMG alginate fragment in the light of its degradation by alginate lyases of Pseudomonas sp; Min KH et al.; An alginate fragment named SMG, consisting of mannuronic (M) and guluronic acid residues (G)(DP=25), was prepared from the partial acid hydrolysate of a commercial alginate . Two subfractions, SMG-ppt (DP=52) and SMG-sup (DP=18) were obtained from SMG by fractionation with MgC12 and CaC12 . The M/G ratios of these alginate fragment were 1.4-1.9 . Their lysis products by a pseudomonad alginate lyase {EC 4.2.2.3} preparation were fractionated by gel filtration, giving similar patterns . The major products in their digests were unsaturated monouronides (53-50%) and triuronides (30-35%) . The former was identified as a delta4,5-hexuronic acid (deltaU) and the latter was identified as a mixture of delta4,5-hexuronosyl-(1 leads to 4)-beta-D-mannuronosyl-(1 leads to 4)-L-guluronic acid (deltaUMG) and delta4,5-hexuronosyl-(1 leads to 4)-alpha-L-guluronosyl-(1 leads to 4))L-guluronic acid (deltaUGG) . The two unsaturated triuronides were present in roughly equal amounts . The presence of 4-O-alpha-L-guluronosyl-L-guluronic acid (GG) and 4-O-beta-D-mannuronosyl-L-guluronic acid (MG) or 4-O-beta-L-guluronosyl-D-mannuronic acid (GM) was also demonstrated inthe digest . Moreover, indirect evidence suggested nonreducing terminal deltaU residue and free deltaU in the digest to be derived more from M than G of the original SMG . Thus, it was concluded that more than one-third of uronic acid residues of SMG molecules may be composed of almost equal amounts of MG and GG sequences, most of which may be connected by M to form MMG and MGG sequences, respectively.

J Biochem (Tokyo), 1977 Mar, 81(3), 547 - 53
Substrate specificity of endo-polyguluronide lyases from Pseudomonas sp . on the basis of their kinetic properties; Min KH et al.; Two endo-alginate lyases {EC 4.2.2.3} differing in their mode of degradation of substrates and practically free of polymannuronide lyase activity were partially purified from Pseudomonas sp . cells . Their substrate specificities were investigated for two different kinds of alginate fragments; a polyguluronide (SG) and a polyuronide consisting of mannuronic (M) and guluronic (G) acid residues (SMG) . The effects of various salts and some organic compounds such as EDTA and p-chloromercuribenzoate on the degradation of the two substrates were similar . High concentrations of the substrates similarly inhibited the action ofthe lyases, giving a bell-shaped plot . A polymannuronide alginate fragment (SM) which was a substrate for polymannuronide lyase but was not attacked by these guluronide lyases also inhibited the degradation of SG and SMG . The overall degradation velocities of a mixture of SG and SMG by both lyases coincided with those calculated from the Michaelis-Menten formula . Based on the above results, it was concluded that SG and SMG are attacked by the same endo-polyguluronide lyase.

J Biochem (Tokyo), 1977 Mar, 81(3), 539 - 46
Multiple components of endo-polyguluronide lyase of Pseudomonas sp; Min KH et al.; A lyophilized alginate lyase preparation obtained from dialyzed extract of sonicated Pseudomonas sp . cells was fractionated by gell filtration on a Sephadex G-150 column, and three alginate lyase {EC4.2.2.3} fractions, peaks I, II, and III, were obtained . They were remarkably thermolabile . The lyase fractions degraded two kinds of alginate fragments, a polygluuronide (SG), and a polyuronide consisting of both mannuronic and guluronic acid residues (SMG), as well as commerical alginate, but were virtually inactive toward polymannuronide fragment (SM) . The modes of degradation of these substrates by the lyase fractions were endowise with different degrees of randomness . Attack by the peak I fraction was more random than those by peaks II and III . The main lysis products formed from SG and SMG by these layses were identified as mixtures of unsaturated tri- and monouronides . The unsaturated triuronide from SG was deltatugg and SMG yielded a mixture of deltaUGG and a poorly characterized unsaturated trimer, possibly deltaUMG . However, the patterns of monomer and trimer production by theselyase fractions changed in different ways during incubation.

Eur J Biochem, 1977 Mar 1, 73(2), 469 - 76
Structure and synthesis of a lipid-containing bacteriophage . Effects of lipids containing cis or trans fatty acids on the reconstitution of bacteriophage PM2; Tsukagoshi N et al.; Infectious PM2 virus paticles could be reconstituted in vitro from a mixture of nucleocapsid, phospholipids containing cis fatty acids, and proteins I and II . The presence or absence of acyl phosphatidylglycerol, a minor lipid component of thevirion, did not affect the reconstitution of infectious particles, even though it was incorporated into the particles when present . When phosphatidylglycerol was completely replaced by acyl phosphatidylglycerol in the reconstitution mixture, no infections particles were formed . Lipids containing either cis or trans fatty acids were also used for reconstitution in vitro of the lipid-containing bacteriophage PM2 . Regardless of the ratio of phosphatidlyglycerol to phospatidylethanolamine in the reconstitution mixture, infectious particles were formed and had almost the same phospholipid composition when lipids containing cis-palmitoleic acid were used; no infectious particles were obtained when lipids containing trans-palmitoleic acid were used . In the latter case, virus-like particles were, however, formed . Reconstituted particles containing cis fatty acids were infectious when tested on wild type Pseudomonas BAL-31 as well as on the unsaturated fatty acid auxotroph grown in the presence of either cis or trans-palmitoleic acid . Reconstituted particles containing trans fatty acids were not infectious on any of these cells . When trans fatty acids as well as cis fatty acids were present in the reconstitution mixture, then there was a lower yield of infectious particles . Particles with either cis or trans fatty acids had all four viral proteins and adsorbed to BAL-31 host cells in a specific manner.

J Bacteriol, 1977 Mar, 129(3), 1379 - 86
Purification and properties of L-glutaminase-L-asparaginase from Pseudomonas acidovorans; Davidson L et al.; An enzyme that catalyzes the hydrolysis of both glutamine and asparagine has been purified to homogeneity from extracts of Pseudomonas acidovorans . The enzyme having a ratio of glutaminase to asparaginase of 1.45:1.0 can be purified by a relatively simple procedure and is stable upon storage . The glutaminase-asparaginase has a relatively high affinity for L-asparagine (Km=1.5 X 10(-5) M) and L-glutamine (Km=2.2 X 10(-5) M) and has a molecular weight of approximately 156,000 the subunit molecular weight being approximately 39,000 . Injections of the enzyme produced only slight increases in the survival time of C3H/HE mice carrying the asparagine-requiring 6C2HED Gardner lymphoma and of white Swiss mice carrying the glutamine-requiring Ehrlich lymphoma.

J Lab Clin Med, 1977 Mar, 89(3), 540 - 3
Myelopoiesis in the infected burn; McEuen DD et al.; A modified bone marrow clonal cell culture technique was used to study granulocyte production during burn injury and sepsis . When rats were inflicted with a 30 percent third-degree scald burn, marrow cellularity and colony-forming units in culture (CFU-C) per 10(5) marrow cells increased progressively to four times normal by 7 days after injury . Conversely, When animals were burned and the burn wound immediately seeded with 10(8) Pseudomonas organisms, CFU-C declined steadily until the day of death and reflected a progressive loss in marrow cellularity . Further studies were conducted replacing or mixing standard colony-stimulating serum with burn, burn-infected, or normal rat serum . The results indicated that colony-stimulating activity could be supplied by postburn serum, but not with normal or burn-infected rat serum . Additionally, serum from burned-infected animals significantly inhibited colony formation when added to the standard colony-stimulating serum . Marrow failure appears to be the major cause for granulocytopenia in burn infection and may partly be serum mediated.

Mikrobiologiia, 1977 Mar-Apr, 46(2), 210 - 6
{Methanol metabolism by Pseudomonas oleovorans}; Loginova NV et al.; A typical facultative methylotroph Pseudomonas oleovorans oxidizes methanol to formaldehyde by a specific dehydrogenase which is active towards phenazine metosulphate . Direct oxidation of formalydehyde to CO2 via formiate is a minor pathway because the activities of dehydrogenases of formaldehyde and formiate are lwo . Most formaldehyde molecules are involved in the hexulose phosphate cycle, which is confirmed by a high activity of hexulose phosphate synthase . Formaldehyde is oxidized to CO2 in the dissimilation branch of the cycle providing energy for biosynthesis; this confirmed by higher levels of dehydrogenases of glucose-6-phosphate and 6-phosphogluconate during the methylotrophous growth of the cells . The acceptor of formaldehyde (ribulose-5-phosphate) is regenerated and pyruvate is synthesized in the assimilation branch of the hexulose phosphate cycle . Aldolase of 2-keto-3-deoxy-6-phosphogluconate plays an important role in this process . Further metabolism of trioses involves reactions of the tricarboxylic acid cycle which performs mainly an anabolic function due to complete repression of alpha-ketoglutarate dehydrogenase during the methylotrophous growth . The carbon of methanol is partially assimilated as CO2 by the carboxylation of pyruvate or phosphoenolpyruvate . NH+4 is assimilated by the reductive amination of alpha-ketoglutarate.

J Pediatr, 1977 Mar, 90(3), 453 - 7
Reservoirs of pseudomonas in an intensive care unit for newborn infants: mechanisms of control; Brown DG et al.; Potential reservoirs of pseudomonas within a neonatal ICU were evaluated . Colonization of infants by the same pseudomonas pyocin types could be classified as a cluster colonization (occurring over three to ten days), or serial colonization (occurring over longer times) . Hands of personnel, sink surfaces, and solutions used to rinse nasopharyngeal catheters were identified as the principle reservoirs . Utilization of a liquid iodophor agent for hand washing and of acetic acid for rinsing suction catheters was associated with a significant reduction in the histologic evidence of sepsis and of pneumonia observed among autopsied infants.

Arch Dermatol, 1977 Feb, 113(2), 199 - 202
Pseudomonas cepacia endocarditis and ecthyma gangrenosum; Mandell IN et al.; A case of right-sided Pseudomonas cepacia endocarditis in a heroin addict is presented in which septic cutaneous vasculitis (ecthyma gangrenosum) is a prominent feature . Ecthyma gangrenosum, most commonly associated with sepsis due to P aeruginosa, has not been previously described with P cepacia septicemia.

J Clin Microbiol, 1977 Feb, 5(2), 167 - 71
Use of stable, sensitized cells in an indirect microhemagglutination test for melioidosis; Hambie EA et al.; Two evaluations were carried out in this study . The first was a comparison of the standard tube test with the automated microtitration test for the detection of antibodies to Pseudomonas pseudomallei by the indirect hemagglutination method . Data from this comparison indicated that the tests were equivalent . The second evaluation consisted of reproducibility studies on two lots of pyruvic aldehyde-stabilized sensitized erythrocytes in comparison with freshly prepared sensitized erythrocytes in the automated microtitration test . The influence of different types of the microtitration plates used was also examined . Results indicated that the use of stabilized antigens is feasible, and these antigens offer the advantage of being ready for immediate use.

Biochem J, 1977 Feb 1, 161(2), 333 - 44
Purification and properties of the methane mono-oxygenase enzyme system from Methylosinus trichosporium OB3b; Tonge GM et al.; 1 . A three-component enzyme system that catalyses the oxidation of methane to methanol has been highly purified from Methylosinus trichosporium . 2 . The components are (i) a soluble CO-binding cytochrome c, (ii) a copper-containing protein and (iii) a small protein; the mol . wts . are 13 000, 47 000 and 9400 respectively . The cytochrome component cannot be replaced by similar cytochrome purified from Pseudomonas extorquens or by horse heart cytochrome c . 3 . The stoicheiometry suggests a mono-oxygenase mechanism and the specific activity with methane as substrate is 6 micronmol/min per mg of protein . 4 . Other substrates rapidly oxidized are ethane, n-propane, n-butane and CO . Dimethyl ether is not a substrate . 5 . The purified enzyme system utilizes ascorbate or, in the presence of partially purified M . trichosporium methanol dehydrogenase, methanol as electron donor but not NADH or NADPH . 6 . Activity is highly sensitive to low concentrations of a variety of chelating agents, cyanide, 2-mercaptoethanol and dithiothreitol . 7 . Activity is highly pH-dependent (optimum 6.9-7.0) and no component of the enzyme is stable to freezing . 8 . The soluble CO-binding cytochrome c shows oxidase acitivity and the relationship between this and the oxygenase activity is discussed.

J Bacteriol, 1977 Feb, 129(2), 821 - 9
Induction and general properties of beta-galactosidase and beta-galactoside permease in Pseudomonas BAL-31; Hidalgo C et al.; The hydrolysis of o-nitrophenyl-beta-D-galactopyranoside (ONPG) by BAL-31, a marine Pseudomonas that acts as a host for bacteriophage PM2, was studied with intact cells and with cell-free extracts . A transport system for ONPG in whole cells and a beta-galactosidase activity in extracts were evident for cells grown on lactose minimal medium . It was found that the addition of isopropylthio-beta-D-galactopyranoside (IPTG) to cells growing in rich medium induced an ONPG hydrolytic activity detectable in cell extracts but cryptic in whole cells . The existence of a transport system for IPTG, which remained cryptic for ONPG, became apparent from studies of the rates of induction of beta-galactosidase as a function of cell mass at different concentrations of IPTG . The main properties of beta-galactosidase and the lactose transport system of BAL-31 were studied in terms of how they were affected by pH, temperature, or by the presence of several sugars . IPTG competitively inhibits the hydrolysis of ONPG by cell extracts . In cells pregrown on lactose, IPTG slightly inhibits the transport of ONPG . Glucose, and with less efficiency lactose, also inhibits the hydrolysis of ONPG in cell extracts . The growth of cells on lactose minimal medium was inhibited by the addition of IPTG . A mechanism for this inhibition and for the inhibition of ONPG transport by IPTG is discussed.

Biochemistry, 1977 Jan 11, 16(1), 100 - 6
Properties of L-methionine gamma-lyase from Pseudomonas ovalis; Tanaka H et al.; The distribution of bacterial L-methionine gamma-lyase (L-methionine methanethiollyase (deaminating) (EC 4.4.1.11) was investigated, and Pseudomonas ovalis (IFO 3738) was found to have the highest activity of enzyme, which was inducibly formed by addition of L-methionine to the medium . L-Methionine gamma-lyase, purified to homogeneity from Ps . ovalis, has a molecular weight of about 173 000 and consists of nonidentical subunits (mol wt: 40 000 and 48 000) . The enzyme exhibits absorption maxima at 278 and 420 nm, and a shoulder around 330 nm, which are independent of the pH (6.0 to 10.0), and contains 4 mol of pyridoxal 5'-phosphate per mol of the enzyme . The formyl group of pyridoxal 5'-phosphate is bound in an aldimine linkage to the epsilon-amino group of lysine residues of the protein . The holoenzyme is resolved to the apoenzyme by incubation with hydroxylamine, and reconstituted by addition of pyridoxal 5'-phosphate . The enzyme activity is significantly affected by both carbonyl and sulfhydryl reagents . L-Methionine gamma-lyase catalyzes alpha,gamma- and alpha,beta-elimination reactions of, in addition to L-methionine, several derivatives of L-methionine and L-cysteine, e.g., L-ethionine, DL-methionine sulfone, L-homocysteine, and S-methyl-L-cysteine . The enzyme catalyzes also gamma-replacement reactions of the thiomethyl group of methionine with various alkanethiols (C2-C7), arylthio alcohols (benzenethiol and beta-naphthalenethiol) and the derivatives of ethanethiol (2-mercaptoethanol and cysteamine) to yield the corresponding S-substituted homocysteine . The thiomethyl group of S-methyl-L-cysteine also is replaced by ethanethiol to form S-ethyl-L-cysteine.

Acta Biochim Pol, 1977, 24(3), 225 - 30
Segregation of the ability to degrade lupanine in Pseudomonas lupanini; Droese J; Two types of mutants differing in the ability to degrade lupanine were obtained from the JD1 strain of Pseudomonas lupanini: lus growing slowly on lupanine, and lun unable to utilize lupanine as a source of carbon and nitrogen . The mutation rate for the spontaneous lus mutants was 0.28%, and for the mitomycin C-induced lus mutants, 3-24% . Rifampicin induced solely lun mutation . The plasmid nature of the lupanine degradation pathway is discussed.

Circ Shock, 1977, 4(2), 201 - 9
Platelets, fibrinogen, and pulmonary haemodynamics in early experimental septic shock; Myrvold HE et al.; The relationship between platelet trapping, fibrinogen, and pulmonary haemodynamics after iv injection of disintegrated Pseudomonas bacteria into dogs were studied . Platelets were labeled with 51Cr and fibrinogen with 125I . The number of circulating platelets and white cells decreased abruptly within 2 minutes after injection, remained low at 5 minutes, and thereafter slowly increased . At the same time there was a transient increase of 51Cr activity in the lung occurring simultaneously with a decrease in cardiac output and an increase in pulmonary vascular resistance . Pulmonary artery pressure remained constant during the first phase of the experiment and thereafter decreased . There were no signs of 125I-fibrinogen accumulation in the lungs during the 2 hours of the experiment . The results indicate that trapping of platelets and eventually leucocytes in the lungs are closely related to the initial pulmonary haemodynamic changes after injection of disintegrated bacteria, possibly both by release of vasoactive substances and mechanical blocking . This microembolism in the pulmonary microcirculation might be of importance for the development of the shock lung syndrome.

J Int Med Res, 1977, 5(5), 308 - 21
Ticarcillin: pharmacokinetics in man according to different administration schedules; Dalhoff A et al.; The pharmacokinetic characteristics of ticarcillin, a semisynthetic penicillin more active than carbenicillin against Pseudomonas, were studied . Following a rapid intravenous infusion of 1 g, 2 g, 5 g and 10 g ticarcillin respectively the serum half-life was 72-4 minutes independent of the dosage administered . If ticarcillin is administered under steady-state conditions, e.g . continuous infusion of either 2g/hr or 1g/hr following a loading dose of 1g (total dose 5 g) the average steady-state serum concentrations are 125 microgram/ml and 105 microgram/ml respectively.

Eur Surg Res, 1977, 9(1), 34 - 47
Microembolism in experimental septic shock . Distribution of platelets and fibrinogen after intravenous injection of disintegrated Pseudomonas bacteria to dogs; Myrvold HE et al.; Platelets were labelled with 51Cr, fibrinogen with 125I and erythrocytes with 59Fe . Disintegrated Pseudomonas bacteria were injected intravenously and radioactive measurements were made on whole blood; tissue biopsies and clottable fibrinogen . After the infection there was an immediate but transient increase of 51Cr activity in the lung concomitant with a decrease in platelet count and 51Cr activity of blood . In the liver there was a less pronounced increase of 51Cr activity . The fibrinogen concentration decreased slightly, paralleled by the 125I activity of whole blood and of clottable fibrinogen, whereas the 125I activity in the lung and liver remained fairly constant . There was no changes of 51Cr activity or 125I activity in biopsies from muscle, pancreas, small intestine, kidney or spleen . During the experiment (3h) there were no signs of significant disseminated intravascular coagulation other than platelet aggregation . A consumption of fibrinogen related to the formation of fibrin plugs could not be detected . After injection of disintegrated Pseudomonas bacteria reversible platelet aggregates were formed and temporarily trapped in the pulmonary microcirculation . This microembolism might induce tissue damage and could be of importance for the development of septic pulmonary complication.

J Infect Dis, 1977 Jan, 135(1), 103 - 7
Wound infection by an indigenous Pseudomonas pseudomallei-like organism isolated from the soil: case report and epidemiologic study; McCormick JB et al.; A 27-year-old farmer in the Oklahoma panhandle was pinned under his overturned tractor for 2 hr and received superficial and deep lacerations . He contracted an infection of a pelvic wound with an organism that had cultural and biochemical characteristics identical to those of Pseudomonas pseudomallei . Identical organisms were recovered from soil taken from the site of the accident . The organism isolated from the wound proved to be less virulent in guinea pigs than usual laboratory strains of P . pseudomallei; fatty acid analysis showed a distinctly different pattern from that of laboratory strains of P . pseudomallei . The infecting organism may be a variant of P . pseudomallei or a new species of Pseudomonas.

J Cardiovasc Surg (Torino), 1977 Jan-Feb, 18(1), 43 - 8
Surgical treatment of acute dissecting aneurysm of the ascending aorta; Seybold-Epting W et al.; Since 1959, 51 patients underwent open heart surgery for correction of an acute dissecting aneurysm of the ascending aorta . Upon admission, 33 patients were severely hypotensive or in progressive heart failure . Acute aortic insufficiency was found in 24 patients, and hemiplegia or hemiparesis in four . In 45 patients the ascending aorta was reconstructed with a woven Dacron graft . After excision of the dissected part of the aorta, primary anastomosis or patch aortoplasty was performed in six patients . The aortic valve remained intact in 26 patients, and resuspension of the commissures restored competence of the aortic valve in another nine . Sixteen patients required aortic valve replacement because of disrupture of the commissures . Dissection extended into the coronary ostia in nine cases . Reconstruction of the coronary system was accomplished by reimplantation of the ostia, interposition of a vein graft or aortocoronary bypass . Nine patients died within the early postoperative course from uncontrollable hemorrhage (four), further dissection (three) and myocardial infarction (two) . Within the first year after surgery, another five patients died from acute aortic dissection (two), pseudomonas infection causing rupture of the proximal graft anastomosis (one) and myocardial infarction (two) . Contraindications of antihypertensive treatment of acute dissection of the ascending aorta are discussed . We recommend prompt surgical intervention in acute dissecting aneurysms of the ascending aorta.

Biochimie, 1977, 59(11-12), 909 - 17
Characterisation of an associate 17-beta-hydroxysteroid dehydrogenase activity and affinity labelling of the 3-alpha-hydroxysteroid dehydrogenase of Pseudomonas testosteroni; Battais E et al.; The 3-alpha-hydroxysteroid dehydrogenase and the 3-beta-hydroxysteroid dehydrogenase of Pseudomonas testosteroni were purified to homogeneity by polyaerylamide gel electrophoresis using the following stages: DEAE cellulose chromatography, affinity chromatography on oestrone-aminocaproate sepharose and Sephadex gel filtration . The pure 3-alpha-hydroxysteroid dehydrogenase was completely devoid of 3-beta-hydroxysteroid dehydrogenase activity but could oxidize estradiol 17-beta at an appreciable rate . This activity accounts for about 40 per cent of the total 17-beta-estradiol dehydrogenase of the crude bacterial extract . Affinity labelling of pure 3-alpha-hydroxysteroid dehydrogenase was carried out using 5-beta-pregnane 3,20-dione-12-alpha-iodoacetate and 5-alpha-androstane 3-one-17-beta-bromoacetate . With both reagents, inactivation was obtained only in the presence of coenzyme, the substrate protected against inactivation and the enzyme was fully inhibited with covalent binding of 1 mole of reagent per mole of subunit suggesting an active site directed inhibition . Histidine and methionine were identified as the labelled aminoacid residues.

Mikrobiyol Bul, 1977, 11(4), 559 - 61
{Pseudomonas-like bacteria (group Ve biotype 1) isolated from a subdiaphragmatic abscess (author's transl)}; Doganay M et al.; Pseudomonas like bacteria group Ve biotype I was isolated three times from the specimens taken by thorocenthessis from a patient with subdiaphragmatic abscess who was admitted to the Thoracic Diseases Clinic in the University of Ankara School of Medicine.

C R Seances Soc Biol Fil, 1977, 171(1), 54 - 9
{Exchange of adenylic nucleotides and bacterial growth}; Champiat D; Adenylic nucleotides pool reaches optimum with biomass . The ATP quantity decreases after exponential growth . AMP increases when ATP decreases . Energetic load is optimum two hours before optimum growth . E . coli prevent the growth of Pseudomonas . AMP is discharged in the medium at the beginning of growth of E . coli and every time by Pseudomonas.

Acta Microbiol Pol, 1977, 26(4), 387 - 92
Humic-like substances of bacterial origin . II . Fractionation of the bacterial humic-like substances by gel filtration on sephadex gels; Kosinkiewicz B; Humic-like substances obtained from cells of Pseudomonas acidovorans were separated on Sephadex G-25 into two groups of substances of different molecular weight . The substances of the molecular weight greater than 5000 were successively separated on Sephadex gels G-50, G-75, G-100 . Five fractions of different molecular weight were obtained, the percentage of which varied depending on the media used and time of incubation of the bacteria . Most (38%--46%) of the compounds contained in the bacterial humic acids were of approximate molecular weight of 40 000--50 000 . The distribution of the fractions in the bacterial "humic-acids" was compared with those of the humic acid made by Fluka A . G . The synthetic humic acid contained most (approximately 40%) of the compounds of approximate molecular weight of 8000--10 000 . In the bacterial and synthetic material the content of the compounds with the molecular weight above 100 000 was very similar (8%--12%).

Acta Microbiol Pol, 1977, 26(4), 377 - 86
Humic-like substances of bacterial origin . I . Some aspects of the formation and nature of humic-like substances produced by Pseudomonas; Kosinkiewicz B; Two bacterial strains Pseudomonas acidovorans No 26 and Pseudomonas sp . No 4 grown in Conn and yeast extract-glucose media, or in the media enriched with tyrosine, were found to produce dark brown pigment . It was shown that in the bacterial cultures numerous phenolic and quinone-type compounds were formed and transformed to humic-like polymers . Formation of humic-like substances started in the bacterial cells and was accompanied by the presence of phenyloxidases in the bacterial cultures . The bacterial "humic acids" were obtained from the supernatants in amounts varing from 0.05 to 0.865 mg/1 mg of dry weight of cells and from the cells in amounts of 0.02 to 0.165 mg/1 mg of dry weight of cells, depending on the medium used and time of incubation . The IR spectra of the bacterial "humic acids" appeared to be very similar to IR spectrum of the synthetic humic acids (Fluka A.G.) and contained the same chemical groups as the soil humic acids . The culture medium after growth of the strain No 26 was fractionated into "fulvic, hymatomelanic and humic acid" fractions . The hydrolysates from the obtained fractions contained amino acids and uronic acids . The amino acid composition appeared to be very similar to that of soil humic acids.

Acta Chem Scand B, 1977, 31(7), 604 - 8
Pseudomonas cytochrome c peroxidase . XIII . pH-denaturation of the enzyme; Soininen R et al.; The effect of pH on the oxidized Pseudomonas cytochrome c peroxidase molecule was studied by measuring the peroxidatic activity, the sedimentation velocity, the circular dichroic spectra in the far UV and Soret regions, and the optical absorption spectra of the enzyme in the pH range 2.5-13.0 at a constant ionic strength (micron = 0.1) . The enzyme was stable in a narrow pH region, pH 6.0 - 7.4 . In the low pH range the gross tertiary structure was observed to change quite simultaneously with the enzymatic activity and secondary structure . The optical absorption spectra indicated that there were no coordinated internal protein liqands in the 6th coordination positions of the heme prosthetic groups at the lowest pH studied . In the high pH range the secondary structure and the protein environment of hemes were observed to remain stable after the tertiary structure had changed and the activity had decreased . According to the optical absorption spectra the 6th internal protein ligands of hemes were retained at the highest pH studied.

Biochim Biophys Acta, 1976 Dec 20, 450(3), 475 - 80
Stabilization of an acetyl-coenzyme A carboxylase complex from Pseudomonas citronellolis; Fall RR; Using stabilizing conditions the acetyl-CoA carboxylase (EC 6.4.1.2) of Pseudomonas citronellolis has been isolated as a complex containing four different polypeptide chains with molecular weights of 53 000, 36 000, 33 000 and 25 000 . Evidence is presented to suggest that these polypeptide chains correspond to distinct biotin carboxylase, transcarboxylase and biotin carboxyl carrier protein subunits in analogy with similar subunits of Escherichia coli acetyl-CoA carboxylase, an unstable complex in vitro.

Biochem J, 1976 Dec 1, 159(3), 707 - 13
Purification and properties of an alginate lyase from a marine bacterium; Davidson IW et al.; An unidentified pseudomonad isolated by enrichment procedures from decomposing seaweed was grown in defined medium containing sodium alginate as the sole carbon source . The alginate lyase recovered from disrupted bacterial cells was purified by a procedure of (NH4)2SO4 precipitation, gel filtration and ion-exchange chromatography . From sodium dodecyl sulphate/polyacrylamide-gel-electrophoresis experiments a mol.wt . of about 50 000 was determined . The enzyme was active against both algal and bacterial alginate preparations . Kinetic studies together with analysis of the unsaturated oligouronide products of alginate lyase action indicated the enzyme was specific for guluronic acid-containing regions of the macromolecular substrate . The specificity of the enzyme can be used to give information about the primary composition of alginate samples.

Am J Dis Child, 1976 Dec, 130(12), 1365 - 6
Treating Pseudomonas cepacia meningitis with trimethoprim-sulfamethoxazole; Darby CP; An infant with neonatal meningitis caused by Pseudomonas cepacia responded promptly to treatment with trimethoprim-sulfamethoxazole (Bactrim) after other abtibiotics had failed . Pseudomonas cepacia has proven to be resistant to most of the commonly used antibiotics.

J Biochem (Tokyo), 1976 Dec, 80(6), 1343 - 52
Superoxide dismutase from Mycobacterium tuberculosis; Kusunose E et al.; 1 . A superoxide dismutase {EC 1.15.1.1} was purified about 275-fold with a yield of 34% from Mycobacterium tuberculosis, strain H37Ra (attenuated strain), grown on a Sauton medium for two months . The purified enzyme was homogeneous as judged by polyacrylamide gel electrophoresis, and by analytical ultracentrifugation and sedimentation equilibrium studies . 2 . The molecular weight of the enzyme was estimated to be approximately 88,000 by sedimentation equilibrium analysis . Since the molecular weight of the subunit was 21,000 as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, the enzyme appears to be composed of four subunits of equal size . 3 . Electron spin resonance (ESR) spectra showed that the enzyme contained ferric iron, and metal analysis showed that the enzyme contained ferric iron, and metal analysis showed that approximately 3.7 atoms of iron were present per mole of the enzyme, indicating the occurrence of 1 atom of iron per subunit . 4 . The amino acid composition was apparently similar to those of the iron-containing superoxide dismutases from Escherichia coli, luminous bacteria, Pseudomonas ovalis, and blue-green alga . 5 . Antibodies against the enzyme were raised in rabbits and immunological studies were performed . The enzyme from M . tuberculosis, strain H37Rv (virulent strain), was found to have antigenic structures identical with those of the H37Ra enzyme . On the other hand, the manganese-containing superoxide dismutases from other species of mycobacteria, i.e., Mycobacterium species, strain Takeo, M . phlei and M . lepraemurium, showed only partial immunological identity with the H37Ra enzyme . 6 . During the growth of M . tuberculosis, strain H37Ra, the enzyme was found to be secreted into the culture medium.

JAMA, 1976 Nov 22, 236(21), 2418 - 9
Contaminated aqueous benzalkonium chloride . An unnecessary hospital infection hazard; Frank MJ et al.; During January and February 1975, nine patients on a single ward of a rural Tennessee hospital unexpectedly developed sepsis . The aseptic technique employed in the management of intravenous infusions was implicated . Pseudomonas cepacia was recovered from the following: bloodstream, inuse intravenous infusions and the antiseptic, aqueous benzalkonium chloride . The outbreak again calls attention to the infection risk associated with the use of this product . Selection of less hazardous antiseptics and disinfectants is strongly recommended.

Biochim Biophys Acta, 1976 Nov 19, 450(2), 131 - 6
Identification of acyl phosphatidylglycerol as a minor phospholipid of Pseudomonas BAL-31; Tsukagoshi N et al.; Compound X, a minor phospholipid of Pseudomonas BAL-31 and bacteriophage PM2, has been identified as X-3-phosphatidyl-1'-(3'-acyl)-glycerol, or acyl phosphatidylglycerol . The water-soluble product obtained by mild alkaline hydrolysis showed the same RF value as that of glycerophosphoryl-glycerol . The chemical analysis gave the ratio 1 : 3 : 2 for phosphate-acyl ester-glycerol . The position of the third acyl group was determined by nuclear magnetic resonance techniques.

Eur J Biochem, 1976 Nov 15, 70(2), 325 - 30
CO2 reduction to formate by NADH catalysed by formate dehydrogenase from Pseudomonas oxalaticus; Ruschig U et al.; The direct reduction of CO2 to formate is catalysed by formate: NAD oxidoreductase in the presence of substrate amounts of NADH . Proof for this reaction is supplied by the detection of a CO2-dependent NADH oxidation, and by the identification of {14c} formate as the product of a NADH-dependent reduction of {14c}carbonate . The enzyme-catalysed CO2 reduction by NADH attains the equilibrium predicted by thermodynamic considerations, a state which is also reached from the formate side . The Michaelis constant for CO2 is about 40 mM indicating the low affinity of the enzyme for this substrate . The corresponding value for formate is 0.1 mM . Under the special conditions employed the enzyme catalyses the formate oxidation about 30 times faster than the CO2 reduction . That CO2 and not HCO3- is the active species in the reduction was shown by comparing the ph dependency of the velocities of the forward and back reactions and by observing the kinetics of CO2 reduction during the simultaneous attainment of the CO2-HCO3- equilibrium.

Vopr Virusol, 1976 Nov-Dec, (6), 724 - 7
{Manifestation of polylysogeny in Pseudomonas vignae}; Kishko IaG et al.; Ultraviolet irradiation and treatment with mitomycin C demonstrated that a culture of Ps . vignae, strain 1025, was polylysogenic, as electron microscopic preparations of phagolysates revealed two types of virus particles: "large" with a hexagonal head of 700 A in diameter, and "small" with a round head of 500 A in diameter . Subcultivation of the test and indicator cultures showed in both cases formation of particles of these phages, the content of "small" virions increasing with the number of passages of negative colonies . Attempts to separate these viruses biological methods were not successfull . The phenomenon of polylysogeny is unusual in that moderate phage adsorbed repeatedly on host bacteria culture and lysed it . The causes of the discovered phenomenon are discussed.

J Gen Microbiol, 1976 Nov, 97(1), 57 - 62
Isolation of pigmentation mutants of Pseudomonas phenazinium; Byng GS et al.; Pigmentation mutants of Pseudomonas phenazinium unable to synthesize iodinin, or producing it only in reduced amounts, were isolated . The abilities of the mutants to synthesize nine other phenazines were also altered . Cross-feeding experiments and the altered patterns of pigment production suggested metabolic relationships between the phenazine pigments, and a scheme for their biosynthesis is proposed.

J Antibiot (Tokyo), 1976 Nov, 29(11), 1147 - 51
Sorbistin, a new aminoglycoside antibiotic complex of bacterial origin . II . Isolation and taxonomy of sorbistin-producing organism; Tomita K et al.; The sorbistin-producing organism Pseudomonas sorbicinii nov . sp . has been isolated from a soil sample by psychrophilic pre-incubation technique . The organism resembles P . fluorescens in many respects but differs in some of the important physiological characteristics such as oxidase production, media specificity for the production of fluorescent pigment, and carbohydrate utilization pattern . The type strain No . D946-B83, has been deposited under the numbers ATCC 31086 and FERM-P 3328.

J Antibiot (Tokyo), 1976 Nov, 29(11), 1137 - 46
Sorbistin, a new aminoglycoside antibiotic complex of bacterial origin . I . Production, isolation and properties; Tsukiura H et al.; A strain of a new Pseudomonas species produced the aminoglycoside antibiotic complex, sorbistin, which was separated by ion-exchange chromatography into three bio-active components A1, A2 and B, and two bio-inactive components C and D . Sorbistins A1, A2 and B showed moderate intrinsic activity against a wide range of bacterial species and inhibited most of the aminoglycoside-resistant organisms . Sorbistin A1 exhibited the highest activity among the three bio-active components . Sorbistins showed low order of acute toxicity in mice.

J Gen Microbiol, 1976 Oct, 96(2), 375 - 81
Properties of phage-receptor lipopolysaccharide from Pseudomonas morsprunorum; Quirk AV et al.; Lipopolysaccharide (LPS) of Pseudomonas morsprunorum was extracted with hot phenol and purified by repeated centrifuging followed by either block electrophoresis or gel filtration . LPS from a virulent isolate exhibited specific phage inactivation (PI50 = 0.05 mug LPS ml-1), whereas LPS from an avirulent phage-resistant mutant did not . LPS was considered pure when a single band was detected following sodium dodecyl sulphate-cellulose acetate electrophoresis (pH 7.4) . It was not phytotoxic when inoculated into cherry leaves at concentrations up to I mg ml-1, but produced weak chlorosis in bean and tobacco at 2 mg ml-1: no visible symptoms appeared after treatment with lower concentrations . The chemical composition of the LPS was partly determined.

J Gen Virol, 1976 Oct, 33(1), 135 - 8
Polyamines in bacteriophage phi W-14 and in phi W-14-infected Pseudomonas acidovorans; Quail A et al.; Bacteriophage phi W-14 is sensitive to osmotic shock . It contain sufficient free putrescine, 2-hydroxyputrescine and spermidine to neutralize about 15% of the DNA phosphates . The alpha-putrescinylthymine residues of the DNA could neutralize a further 25% of the phosphates . Label is transferred from ornithine to the alpha-putrescinyl residues of phi W-14 DNA . The rates of polyamine synthesis in Pseudomonas acidovorans are increased by phi W-14 infection.

Can J Microbiol, 1976 Oct, 22(10), 1577 - 9
Adsorption of Pseudomonas pseudomallei cytophilic antibodies onto rabbit alveolar macrophages; Kishimoto RA et al.; Cytophilic antibody has been demonstrated in sera from rabbits immunized with Pseudomonas pseudomallei . This antibody passively adsorbed onto normal rabbit alveolar macrophages rendered the macrophages agglutinable by P . pseudomalei antigens and capable of displaying 'rosette' formation with polysaccharide-sensitized erythrocytes.

Eur J Biochem, 1976 Sep 15, 68(2), 323 - 31
Purification and properties of L-4-hydroxymandelate oxidase from Pseudomonas convexa; Bhat SG et al.; An inducible membrane-bound L-4-hydroxymandelate oxidase (decarboxylating) from Pseudomonas convexa has been solubilized and partially purified . It catalyzes the conversion of L-4-hydroxymandelic acid to 4-hydroxybenzaldehyde in a single step with the stoichiometric consumption of O2 and liberation of CO2 . The enzyme is optimally active at pH 6.6 and at 55 degrees C . It requires FAD and Mn2+ for its activity . The membrane-bound enzyme is more stable than the solubilized and purified enzyme . After solubilization it gradually loses its activity when kept at 5 degrees C which can be fully reactivated by freezing and thawing . The Km values for DL-4-hydroxymandelate and FAD are 0.44 mM and 0.038 mM respectively . The enzyme is highly specific for DL-4-hydroxymandelic acid . DL-3,4-Dihydroxymandelic acid competitively inhibited the enzyme reaction . From the Dixon plot the Ki for DL-3,4-dihydroxymandelic acid was calculated to be 1.8 X 10(-4) M . The enzyme is completely inactivated by thiol compounds and not affected by thiol inhibitors . The enzyme is also inhibited by denaturing agents, heavy metal ions and by chelating agents.

J Biol Chem, 1976 Sep 10, 251(17), 5375 - 80
Physical properties of antitumor glutaminase-asparaginase from Pseudomonas 7A; Holcenberg JS et al.; Glutaminase-asparaginase from Pseudomonas 7A appears to have four subunits with a molecular weight of 36,000 +/- 500 by sedimentation equilibrium in 5.9 M guanidine HCl and 34,000 by amino acid analysis . Analytic sedimentation equilibrium of the native enzyme showed a molecular weight of 140,000 +/- 3,300 with no signs of association or dissociation . Moving boundary and zone sedimentation in buffer showed normal behavior with sedimentation coefficients of 7.92 and 7.75 S, respectively . In contrast, the enzyme appeared to polymerize during zone sedimentation when the initial protein concentration was greater than 1 mg/ml and the buffer contained asparagine, glutamine, or 5-diazo-4-oxonorvaline . An extension of our method for active enzyme sedimentation is described which utilizes the changes in absorption during hydrolysis of asparagine by low concentrations of enzyme . Polymerization was not seen under these conditions.

Southeast Asian J Trop Med Public Health, 1976 Sep, 7(3), 363 - 6
Pseudomonas putrefaciens from clinical material; Thong ML; Three strains of Pseudomonas putrefaciens were isolated from routine clinical specimens at the University Hospital, Kuala Lumpur, Malaysia . Their cultural and biochemical characteristic, and antibiotic susceptibilities are presented . Characteristics of diagnostic value were stressed . Two isolates appeared to have played a pathogenic role in chronic otitis media.

J Gen Microbiol, 1976 Sep, 96(1), 17 - 24
Sodium-dependent growth and respiration of a nonhalophilic bacterium, Pseudomonas stutzeri; Kodama T et al.; Pseudomonas stutzeri (van Niel strain) requires Na+ for growth . Its growth rate was a sigmoidal function of Na+ concentration, being maximal and constant from 2 to 50 mM-Na+, and half maximal at about 0-5 mM-Na+ . The relationship between cell concentration and Na+ concentration was non-linear; cell concentration increased abruptly when Na+ was greater than 0-3 mM . Accumulation of Na+ in the organism during growth was not detected . In the presence of K+, respiration was enhanced specifically by Na+ . The respiration rate of the organism growing in the culture was a linear function of the growth rate when limited by the Na+ concentration, whereas the maximum rate induced by excess Na+ was independent of the growth rate.

Laryngoscope, 1976 Sep, 86(9), 1386 - 90
Pseudomonas meningitis complicating head and neck surgery; Bray DA et al.; Three patients, each of whom had Pseudomonas meningitis as a sequela of an extensive head and neck operation, have been treated successfully . All three patients had cerebrospinal fluid leaks, and operative management of this complication is discussed . Antibiotic management included the parenteral administration of the recently developed drugs gentamicin, carbenicillin, and intrathecal gentamicin . Since extensive head and neck operations are being performed, with increasing frequency and since infectious complications are inevitable, it is mandatory that the otolaryngologist be familiar with current methods of managing these potentially lethal conditions.

J Pediatr, 1976 Sep, 89(3), 382 - 7
Severe combined immunodeficiency with leukopenia (reticular dysgenesis) in siblings: immunologic and histopathologic findings; Ownby DR et al.; The hematologic and histologic features of two, nontwin, male siblings with severe combined immunodeficiency and variable granulocytopenia are compared to the four previously reported cases of reticular dysgenesis . These sibs died at 50 and 3 days of age, respectively, with Pseudomonas sepsis and congenital cytomegalovirus infection, respectively . A maternal uncle has selective IgA deficiency . Cord blood from the second sib contained a normal percentage of E-rosetting lymphocytes; however, these lymphocytes failed to respond to mitogenic stimulation in vitro . Erythrocyte and lymphocyte levels of adenosine deaminase were elevated in the father and the second sib . Serum immunoglobulin concentrations were low in both siblings.

J Bacteriol, 1976 Sep, 127(3), 1217 - 24
XYL, a nonconjugative xylene-degradative plasmid in Pseudomonas Pxy; Friello DA et al.; Pseudomanas Pxy metabolizes p- or m-xylene through intermediate formation of the corresponding methylbenzyl alcohol and toluic acid via the meta pathway . The strain Pseudomonas Pxy spontaneously loses its ability to grow with xylene or toluate, and the rate of loss of this ability is greatly enhanced by treatment of the cells with mitomycin C . The assay of enzymes involved in xylene degradation in xylene-negative Pxy cells indicates the loss of the entire enzyme complement of the pathway . The genes specifying all the xylene-degradative enzymes, including those of the meta pathway, appear to be borne on a nonconjugative plasmid and can be transferred to xylene-negative Pxy or P . putida strain PpG1 cells only in the presence of a transfer plasmid termed factor K . When transferred to strain PpG1, the xylene-degradative plasmid, termed XYL, coexists stably with factor K, but transduction of XYL is not accompanied by a cotransfer of factor K . XYL appears to be compatible wit- all the other known degradative plasmids in P . putida . The xylene pathway is inducible in wild-type Pxy as well as in Pxy and PpG1 exconjugants, suggesting the cotransfer of regulatory genes along with the plasmid . The enzymes converting xylene to toluate are induced by xylene, methylbenzyl alcohol, or the aldehyde derivatives but not significantly by toluate, whereas catechol dioxygenase and other enzymes are induced by toluates and presumable by xylene as well.

Mikrobiologiia, 1976 Sep-Oct, 45(5), 787 - 90
{Effect of dissolved oxygen concentration on the growth of Pseudomonas}; Matiashova RN; The effect of concentration of oxygen dissolved in the medium (O2) on growth was studied with Pseudomonas herbicola, Ps . non-liquefaciens, Ps . aeruginosa, and Ps . desmolytica . Growth of all studied Pseudomonas cultures was decelerated when (O2) fell beyond 30-50% of saturation of the medium with oxygen; (O2) critical for respiration was within the range of 11-17% saturation, i . e . was 2-3 times lower that for growth . A decrease in the specific growth rate was usually accompanied with a decrease of the economic coefficient.

Z Naturforsch {C}, 1976 Sep-Oct, 31(9-10), 601 - 5
{Chromosomal structure of Pseudomonas testosteroni . II . Activity of the endogenous RNA-polymerase (author's transl)}; Reimer G et al.; After careful lysis the nucleoid of Pseudomonas testosteroni can be isolated in three different forms with compact and unfolded DNA structures . The released nucleoids contain endogenous DNA-dependent RNA-polymerase activity using the chromosomal DNA as a template . RNA synthesis is proportional to duration of RNA-polymerase reaction and amount of DNA-protein-complexes . The sensitivity towards ionic strength and rifampicin indicates that a part of RNA-polymerase activity is tightly bound to the chromosomal DNA.

Prikl Biokhim Mikrobiol, 1976 Sep-Oct, 12(5), 704 - 8
{Purification and properties of L-glutamine and L-asparagine deaminase from Pseudomonas aurantiaca IBPM-14}; Vinogradov BD et al.; An enzymic preparation of L-glutamine and L-asparagine deamidase was obtained from Pseudomonas auractiaca IBPM B-14 . The preparation was purified 100--150-fold by thermal treatment and chromotography on columns with biogel P-150 and DEAE-cellulose . The enzymic activity was measured by the methods of hydroxylaminolysis and direct nesslerization . The deamidase preparation had an activity of 51 i . u . by glutamine and 15 i . e . by asparagine . Evidence on the pH effect on the deamidase activity was accumulated.

J Gen Microbiol, 1976 Sep, 96(1), 185 - 93
Purification and characterization of phosphoglycerate mutase from methanol-grown Hyphomicrobium X and Pseudomonas AM1; Hill B et al.; Phosphoglycerate mutase has been purified from methanol-grown Hyphomicrobium X and Pseudomonas AMI by acid precipitation, heat treatment, ammonium sulphate fractionation, Sephadex G-50 gel filtration and DEAE-cellulose column chromatography . The purification attained using the Hyphomicrobium X extract was 72-fold, and using the Pseudomonas AMI extract, 140-fold . The enzyme purity, as shown by analytical polyacrylamide gel electrophoresis, was 50% from Hyphomicrobium X and 40% from Pseudomonas AMI . The enzyme activity was associated with one band . The purified preparations did not contain detectable amounts of phosphoglycerate kinase, phosphopyruvate hydratase, phosphoglycerate dehydrogenase or glycerate kinase activity . The molecular weight of the enzymic preparation was 32000 +/- 3000 . The enzyme from both organisms was stable at low temperatures and, in the presence of 2,3-diphosphoglyceric acid, could withstand exposure to high temperatures . The enzyme from Pseudomonas AMI has a broad pH optimum at 7-0 to 7-6 whilst the enzyme from Hyphomicrobium X has an optimal activity at pH 7-3 . The cofactor 2,3-diphosphoglyceric acid was required for maximum enzyme activity and high concentrations of 2-phosphoglyceric acid were inhibitory . The Km values for the Hyphomicrobium X enzyme were: 3-phosphoglyceric acid, 6-0 X 10(-3) M: 2-phosphoglyceric acid, 6-9 X 10(-4) M; 2,3-diphosphoglyceric acid, 8-0 X 10(-6) M; and for the Pseudomonas AMI ENzyme: 3-4 X 10(-3) M, 3-7 X 10(-4) M and 10 X 10(-6) M respectively . The equilibrium constant for the reaction was 11-3 +/- 2-5 in the direction of 2-phosphoglyceric acid to 3-phosphoglyceric acid and 0-09 +/- 0-02 in the reverse direction . The standard free energy for the reaction proceeding from 2-phosphoglyceric acid to 3-phosphoglyceric acid was -5-84 kJ mol(-1) and in the reverse direction +5-81 kJ mol(-1).

J Bacteriol, 1976 Sep, 127(3), 1197 - 1207
Isolation and characterization of dihydropteridine reductase from Pseudomonas species; Williams CD et al.; Dihydropteridine reductase isolated from the bacterium Pseudomonas species (ATCC 11299a) has been purified approximately 450-fold byammonium sulfate precipitation and diethylaminoethyl-cellulose chromatographic procedures . The preparation is at least 80% pure as judged by polyacrylamide gels . Its molecular weight was determined to be about 44,000 . Both dihydropteridine reductase and phenylalanine hydroxylase activities were found to be higher in cells adapted to a medium containing L-phenylalanine or L-tyrosine as the sole carbon source than in those grown in L-asparagine . The substrate of the reductase is quinonoid dihydropteridine, and the product is tentatively identified as a tetrahydropteridine through its ability to serve as a cofactor for phenylalanine hydroxylase . The enzyme shows no marked specificity for the pteridine cofactor that occurs naturally in this organism, L-threo-neopterin . The pH optimum for the reductase is 7.2, and nicotinamide adenine dinucleotide, reduced form, is the preferred cosubstrate . Inhibition of the reduced and untreated enzyme by several sulfhydryl reagents was observed . A metal requirement for the reductase could not be demonstrated . Dihydropteridine reductase was found to be inhibited by aminopterin in a competitive manner with respect to the quinonoid dihydro form of 2-amino-4-hydroxy-6,7-dimethyl-5,6,7,8-tetrahydropteridine.

Biochem J, 1976 Aug 15, 158(2), 223 - 9
The biochemical pathway for the breakdown of methyl cyanide (acetonitrile) in bacteria; Firmin JL et al.; {2-14C}Methyl cyanide (acetonitrile) is metabolized to citrate, succinate, fumarate, malate, glutamate, pyrrolidonecarboxylic acid and aspartate . Non-radioactive acetamide and acetate compete with 14C from methyl cyanide, and {2-14C}acetate and {2-14C}methyl cyanide are metabolized at similar rates, giving identical products . This evidence, combined with the inhibitory effect of fluoroacetate and arsenite on methyl cyanide metabolism, indicates that the pathway is: methyl cyanide leads to acetamide leads to acetate leads to tricarboxylic acid-cycle intermediates . The pathway was investigated in a species of Pseudomonas (group III; N.C.I.B . 10477), but comparison of labelling patterns suggests that it also exists in several higher plants.

J Biol Chem, 1976 Aug 10, 251(15), 4792 - 3
Crystallization and preliminary crystal data of iron-containing superoxide dismutase from Pseudomonas ovalis; Yamakura F et al.; Large single crystals of iron-containing superoxide dismutase from Pseudomonas ovalis were prepared from 50% saturated ammonium sulfate solution, at pH 4.5, on gentle evaporation of the solvent at 4 degrees . The crystals were monoclinic, space group P2, with unit cell dimensions a = 81.9 A, b = 49.0 A, c = 61.0 A, and beta = 106 degrees . Considerations of cell volume and protein molecular weight indicated 1 molecule of superoxide dismutase in the assymmetric unit, the smallest number reported so far for superoxide dismutases.

Biochemistry, 1976 Aug 10, 15(16), 3465 - 72
Multiple acyl-coenzyme A carboxylases in Pseudomonas citronellolis; Hector ML et al.; Pseudomonas citronellolis was shown to contain four different acyl-coenzyme A carboxylases, including acetyl-, propionyl-, 3-methylcrotonyl-, and geranyl-CoA carboxylases, when grown on the appropriate carbon sources . Acetyl-CoA carboxylase activity in crude extracts was stimulated approximately 40-fold by inclusion of 0.4-0.5 M ammonium sulfate in the assay . Unexpectedly high levels of propionyl-CoA carboxylase activity, also stimulated by ammonium sulfate, were found in acetate-grown cells . That these acetyl- and propionyl-CoA carboxylase activities were due to different enzymes was shown by their resolution during purification by a procedure that stabilized acetyl-CoA carboxylase as a complex and separated propionyl-CoA carboxylase into two required protein fractions . Propionate- or valine-grown cells contained a propionyl-CoA carboxylase activity that was strongly inhibited by ammonium sulfate in the assay, and which may represent an inducible form of the enzyme . Geranyl- and 3-methylcrotonyl-CoA carboxylases that catalyze the carboxylation of the 3-methyl groups of homologous acyl-CoA acceptors, were induced by growth on the monoterpenes, citronellic or geranoic acid; only 3-methylcrotonyl-CoA carboxylase was induced by growth on leucine or isovaleric acid . Induction of either carboxylase was associated with the appearance of similar high-molecular-weight, biotin-containing proteins as measured by gel filtration . These two carboxylases are probably distinct enzymes since 3-methyl-crotonyl-CoA carboxylase from isovalerate-grown cells does not carboxylate geranyl-CoA, while geranyl-CoA carboxylase will carboxylate both acyl-CoA homologues . P . citronellolis appears to be a useful system for studying the structural aspects of pairs of homologous acyl-CoA carboxylases.

Acta Ophthalmol (Copenh), 1976 Aug, 54(4), 401 - 7
A case of late pseudomonas ocular infection following scleral buckling; Romem M et al.; Two and half years after a circling buckle operation in which two Supra-mid sutures and a solid silicone explant were employed, a Pseudomonas infection developed around the explant and progressed to the inner eye . Removal of all implanted foreign material and treatment with Rifamicin saved the eye . The relevant literature on infection following sclero-plastic procedures is reviewed, and the pathogenesis of the late infections is discussed.

Am J Optom Physiol Opt, 1976 Aug, 53(8), 431 - 2
Preventable hazard of soft lens wear; Dada VK et al.; Recurrent pseudomonal conjunctivitis, caused by poor soft contact lens care and the use of contaminated saline, is described . This problem can easily be avoided by proper sterilization, with boiling being the method of choice.

Appl Environ Microbiol, 1976 Aug, 32(2), 213 - 6
Metabolism of DDT analogues by a Pseudomonas sp; Francis AJ et al.; A Pseudomonas sp . rapidly metabolized several nonchlorinated analogues of DDT, with the exception of 2,2-diphenylethanol, as the sole carbon source . Several of the mono-p-chloro-substituted diphenyl analogues were also metabolized as the sole carbon source by the bacterium . The resulting chlorinated aromatic acid metabolites were not further metabolized . The isolate was unable to metabolize p,p'-dichlorodiphenyl analogues as the sole carbon source.

Can J Microbiol, 1976 Aug, 22(8), 1206 - 8
The relationship between chemical structure of attractants and chemotaxis by a marine bacterium; Chet I et al.; The chemotactic responses of a marine pseudomonad to steroisomers and analogues of amino acids and sugars were tested . The data reveal that the bacterium is equally attracted to D, L, and DL forms of the amino acids . In contrastr, chemical analogues of the amino acids and glucose yielded significantly lower chemotactic responses . The threshold of bacterial detection was 10(-8) M for leucine and cysteine . However, the threshold molarity of most of the analogues was higher than those of the related amino acids and sugars.

J Virol, 1976 Aug, 19(2), 446 - 56
Phospholipid metabolism in Pseudomonas BAL-31 infected with lipid-containing bacteriophage PM2; Diedrich DL et al.; Infection of Pseudomonas BAL-31 with the lipid-containing bacteriophage PM2 resulted in no detectable change in the rate of phosphatidylglycerol (PG) or phosphatidylethanolamine (PE) biosynthesis . An increase in the PG content of infected cultures was not seen until the cultures began to lyse, and this increase was in fact only a relative increase resulting from the extensive turnover of PE at the onset of culture lysis . Turnover studies revealed that the glycerol, phosphorus fatty acid, and ethanolamine moieties of PE turned over simultaneously at the time of lysis, and therefore made it unlikely that there was a PE to PG conversion during the latent period of the phage . The lipid found in the bacteriophage did not reflect a preferential selection for lipid synthesized before or after infection, but in fact reflected the composition of the host membrane at the time the phage were assembled . The use of a modified medium that allowed the cultivation of Pseudomonas BAL-31 as a prototroph and resulted in reliable lysis times of infected cultures led us to the conclusion that PM2 infection effects little change in host phospholipid metabolism, and that there is sufficient PG in the host cytoplasmic membrane to account for a full burst of phage . As a result of the reliable lysis times that we have achieved, we concluded that certain metabolic events, i.e., PE turnover, are lytic phenomena and must not be confused with events relevant to the biosynthesis and maturation of the phage.

J Biochem (Tokyo), 1976 Aug, 80(2), 361 - 6
Studies on phospholipase C from Pseudomonas aureofaciens . II . Further studies on the properties of the enzyme; Sonoki S et al.; Phospholipase C {EC 3.1.4.3} from Pseudomonas aureofaciens was found to be inhibited by chelating reagents such as ethylenediaminetetraacetate {EDTA} and o-phenanthroline . The inhibition was reversed by the addition of Zn2+ and, to a lesser extent, by Co2+ and Mn2+ . On isoelectric focusing, the isoelectric point of this enzyme proved to be 6.3--6.5, with a single peak . The enzyme reaction with the substrate was followed in media containing an organic solvent such as diethyl ether or diethyl ether-ethyl alcohol . When ethyl alcohol was added (up to 2%) to the reaction mixture in ether, there were no marked changes in the hydrolytic rates of phosphatidylcholine and phosphatidylethanolamine . However, the enzyme activity was inhibited when the alcohol concentration was increased above 2% . In 98% diethyl ether-2% ethyl alcohol, phosphatidylcholine was hydrolyzed more rapidly than phosphatidylethanolamine, in contrast with the result obtained in water . In the single micelle state, phosphatidylethanolamine was hydrolyzed more rapidly than phosphatidylcholine or lysophatidylcholine . Acidic phospholipids and sphinogomyelin were not hydrolyzed . When the enzyme was incubated with phospholipid mixture extracted from Ps aureofaciens and rat liver, both phosphatidylethanolamine and phosphatidylcholine were hydrolyzed more rapidly than in the single micelle state of these substrates.

Gastroenterology, 1976 Aug, 71(2), 365 - 8
Bacterial variants: etiologic agent in Crohn's disease?
Parent K, Mitchell PD.
Bacterial variants of Pseudomonas maltophilia and Pseudomonas-like bacteria were recovered from tissues removed during the surgical treatment of three successive patients with Crohn's disease and from one patient with clinical and pathological features of both Crohn's disease and chronic ulcerative colitis . Bacterial variants were not cultured from colonic specimens of one patient with classical features of chronic ulcerative colitis and two patients with adenocarcinoma of the colon . The findings suggest that a relationship exists between variant bacteria and the pathogenesis of Crohn's disease.

Biochem J, 1976 Aug 1, 157(2), 431 - 8
Some spectral and steady-state kinetic properties of Pseudomonas cytochrome oxidase; Barber D et al.; Some spectra of Pseudomonas cytochrome oxidase are reported, both for comparison with those of other workers and to illustrate the differences between the ascorbate- and dithionite-reduced forms of the enzyme . A spectrum of the reduced enzyme-CO complex, prepared in the absence of added reductants by incubation under CO, is also included . Ultracentrifugation studies yielded a value for the sedimentation coefficient (s20,w) of 7.5S, and an isoelectric point of pH6.9 was determined by isoelectric focusing . Steady-state kinetic constants of the electron donors, quinol, sodium ascorbate, reduced Pseudomonas azurin and Pseudomonas ferrocytochrome c551 were investigated giving Km values of 30mM, 4mM, 49muM and 5.6muM respectively . The two protein substrates were observed to be subject to product inhibition and the Ki for oxidized Pseudomonas azurin was evaluated at 4.9muM . Steady-state kinetics were also used to investigate the effects of the oxidation products of dithionite on the oxidase and nitrite reductase activities of Pseudomonas cytochrome oxidase . These experiments showed that whereas the oxidase activity was inhibited, the nitrite reductase activity was slightly enhanced.

Mikrobiologiia, 1976 JUL-AUG, 45(4), 650 - 4
{para-Aminobenzoic acid as a sole source of carbon and energy for Pseudomonas desmoliticum}; Surovtseva EG et al.; A strain of Psuedomonas desmoliticum has been isolated from soil . It utilizes, as a source of carbon and energy, p-aminobenozic (PABA), p-fluorobenzoic acids, and some other aromatic compounds . The strain has been isolated by inoculating soil suspensions onto Petri plates with a solid mineral medium containing 0.1% PABA as a carbon source . The preparatory metabolism of PABA was studied in this work; p-hydroxybenzoic and protocatechuic acids were found to be its intermediate products . Enzyme systems catalysing oxidation of aromatic compounds and glucose are inducible.

Prikl Biokhim Mikrobiol, 1976 Jul-Aug, 12(4), 524 - 7
{New strain of methylotrophic bacteria Pseudomonas oleovorans--a polysaccharide producer}; Egorov AM et al.; A new strain of facultative-methylotrophic bacteria was isolated and identified as Pseudomonas oleovorans . The culture grew on the methanol-containing medium (up to 6%) . In addition to methanol, bacterial growth was provided by different organic acids, multicarbon alcohols and carbohydrates . During growth bacteria synthesized extracellular polysaccharide (up to 400 mg/ml) . The compound included glucose, galactose and xylose.

Zentralbl Bakteriol {Orig B}, 1976 Jul, 162(1-2), 169 - 79
{Primary oxidation mechanisms in degradation of aliphatic hydrocarbons by bacterial enzyme systems (author's transl)}; Hammer KD et al.; The bacterial dissimilation of aliphatic hydrocarbons is catalysed by a monooxygenase mechanism with incorporation of molecular oxygen . Numerous publications have shown the cytochrome P 450-dependent hydroxylation of hydrocarbons, but there is considerably less information of hemo-protein-independent hydroxylations by alkanhydroxylases . In a marine Pseudomonad we found a system sensitive to cyanide: The oxygenase could be divided into three protein fractions . A cytochrome P 450 type spectrum was not detected . The NADH-dependent hydroxylation of n-decane can be activated by Mg2+ and Fe2+ ions . A noncompetitive product inhibition occurs which deserves special attention . An alcohol-dehydrogenase is closely associated with the oxygenase system by a kind of multienzyme-complex . Studies on kinetics and substrate specificity of this enzyme show an inhibition by excess substrate increasing with the chain length of the alcohols . The whole complex (alkanhydroxylase, alcoholdehydrogenase and aldehyddehydrogenase) is induceable by bacterial growth on alkanes, primary alcohols and fatty acids as sole carbon source.

Zentralbl Bakteriol {Orig B}, 1976 Jul, 162(1-2), 127 - 37
{On the mechanism of the biological persistence of halogenated and sulfonated aromatic hydrocarbons (author's transl)}; Knackmuss HJ et al.; Aromatic compounds with unphysiological substituents like halogen or SO2H-groups are mainly degraded by cometabolism . Investigations with model compounds show, that the negative inductive effect (-I-effect) of the halogen substituents impede the electrophilic attack of the oxygenases, particularly the pyrocatechases . Although the "affinity" of the enzyme for the substrate increases with the number of halogen substituents, the rate of ring cleavage decreases by the presence of halogen substituents . Benzoate oxygenation shows that in certain positions of substitution the -I-effect can be weakened by the + M-effect of the halogen . As soon as aromaticity is lost by the action of dioxygenases, halide can be eliminated with greater ease and total mineralization is possible . When naphthalene-2-sulfonic acid is degraded by a naphthalene utilizing Pseudomonas strain the sulfonic acid group appears to be eliminated oxygenolytically and the carbon skeleton is channelled into the naphthalene pathway.

Biochem J, 1976 Jul 1, 157(1), 197 - 205
A steady-state kinetic study of the reaction catalysed by the secondary-amine mono-oxygenase of Pseudomonas aminovorans; Brook DF et al.; 1 . Secondary-amine mono-oxygenase (proposed EC group 1.14.99.-) was partially purified from trimethylamine-grown Pseudomonas aminovorans by (NH4)2SO4 fractionation, gel filtration, hydrophobic chromatography on 5-aminopentylamino-Sepharose, and affinity chromatography on Sepharose-bound NADH . 2 . Some problems in the affinity-chromatography step are discussed . 3 . A steady-state kinetic analysis varying substrate, oxygen and electron-donor concentrations was performed, which, over the concentration range studied, gave a series of families of approximately parallel double-reciprocal plots . From secondary and tertiary plots, Michaelis constants of 0.160 mM, 0.086 mM and 0.121 mM were obtained for dimethylamine, NADPH and oxygen respectively . 4 . Product-inhibition studies supported the postulated Hexa Uni Ping Pong (triple-transfer) reaction mechanism.

J Gen Microbiol, 1976 Jul, 95(1), 121 - 33
Synthesis and hydrolysis of malyl-coenzyme A by Pseudomonas AM1: an apparent malate synthase activity; Cox RB et al.; The malate synthase activity detectable in crude extracts of Pseudomonas AM1 has been shown to be due to a coupling of a malyl-CoA hydrolase with malyl-CoA lyase and not due to a discrete malate synthase enzyme . The partial purification of this malyl-CoA hydrolase from Pseudomonas AM1 has shown that it is distinct from citrate synthase which also hydrolyses malyl-CoA . The malyl-CoA hydrolase has a low Km for malyl-CoA (7-0 muM) . A mutant of Pseudomonas AM1, ICT51 (Taylor & Anthony, 1975), which is unable to grow on ethanol, malonate or 3-hydroxybutyrate, has been shown to have an altered malyl-CoA hydrolase with a Km for malyl-CoA 30 times higher than that of the enzyme present in the wild-type organism . Two classes of revertants to growth on these substrates have been isolated: (i) those with a malyl-CoA hydrolase of similar Km to the wild-type and (ii) those in which the malyl-CoA hydrolase activity remains the same as in the mutant ICT51 . The nature of the mutation leading to the latter class of revertants is unknown.

J Biochem (Tokyo), 1976 Jul, 80(1), 79 - 87
Purification and properties of an enzyme catalyzing the splitting of carbon-mercury linkages from mercury-resistant Pseudomonas K-62 strain . I . Splitting enzyme 1; Tezuka T et al.; An enzyme (S-1) which catalyzes the splitting of carbon-mercury linkages of organomercury compounds was purified about 24-fold from the cell-free extract of mercury-resistant Pseudomonas K-62 strain by treatment with streptomycin, precipitation with ammonium sulfate, and successive chromatography on Sephadex G-150, DEAE-Sephadex, and DEAE-cellulose . A purified preparation of the enzyme showed a single band on polyacrylamide gel electrophoresis, and was colorless . The molecular weight of the enzyme was estimated to be 19,000, and Km was 5.3 X 10(-5) M for p-chloromercuribenzoic acid (PCMB) . The temperature and pH optimum for the reaction were 50degrees and 7.0, respectively . The enzyme was capable of catalyzing the decomposition of methylmercuric chloride (MMC), ethylmercuric chloride (EMC), phenylmercuric acetate (PMA), and PCMB in the presence of a sulfhydryl compound to form a mercuric ion plus methane, ethane, benzene, or benzoic acid, respectively . The mercuric ion thus formed was reduced to metallic mercury by metallic mercury-releasing enzyme (MMR-enzyme).

J Gen Microbiol, 1976 Jul, 95(1), 134 - 43
Acetyl-CoA production and utilization during growth of the facultative methylotroph Pseudomonas AM1 on ethanol, malonate and 3-hydroxybutyrate; Taylor IJ et al.; In Pseudomonas AM1, conversion of 3-hydroxybutyrate to acetyl-CoA is mediated by an inducible 3-hydroxybutyrate dehydrogenase, an acetoacetate: succinate coenzyme A transferase (specific for succinyl-CoA) and an inducible beta-ketothiolase . Ethanol is oxidized to acetate by the same enzymes as are involved in methanol oxidation to formate . An inducible acetyl-CoA synthetase has been partially purified and characterized; it is essential for growth only on ethanol, malonate and acetate plus glyoxylate, as shown by the growth characteristics of a mutant (ICT54) lacking this enzyme . Free acetate is not involved in the assimilation of acetyl-CoA, and hydroxypyruvate reductase is not involved in the oxidation of acetyl-CoA to glyoxylate during growth on 3-hydroxybutyrate . A mutant (ICT51), lacking 'malate synthase' activity has been isolated and its characteristics indicate that this activity is normally essential for growth, of Pseudomonas AM1 on ethanol, malonate and 3-hydroxybutyrate, but not for growth on other substrates such as pyruvate, succinate and C1 compounds . The growth properties of a revertant (ICT51R) and of a mutant lacking malyl-CoA lyase (PCT57) indicate that an alternative route must exist for assimilation of compounds metabolized exclusively by way of acetyl-CoA.

Biochim Biophys Acta, 1976 Jun 7, 438(1), 13 - 22
Affinity chromatography of 3 alpha-hydroxysteroid dehydrogenase from Pseudomonas testosteroni . Use of N,N-dimethylformamide to prevent hydrophobic interactions between the enzyme and the ligand; Aukrust LE et al.; 1 . The 3alpha-hydroxysteroid: NAD+-oxidoreductase (EC 1.1.1.50) from Pseudomonas testosteroni (ATCC 11996) has been purified by affinity chromatography on Sepharose 4B using glycocholic acid as ligand covalently bound through its carboxyl group to the ethylenediamine spacer . 2 . The attachment of the enzyme to the substrate-containing matrix is greatly enhanced by the presence of NAD+ suggesting that this enzyme has a compulsory ordered mechanism where NAD+ binds to the enzyme before the steroid . 3 . A NAD+-independent interaction between the enzyme and the ligand was also found . This interaction was mainly hydrophobic and interfered with the NAD+-dependent binding . The NAD+-independent interaction was reduced by N,N-dimethylformamide . 4 . By using the affinity column in the presence of 10% N,N-dimethylformamide, highly purified enzyme, as judged from polyacrylamide gel electrophoresis, could be obtained in one step from crude bacterial extracts.

Biochemistry, 1976 Jun 1, 15(11), 2323 - 7
Isobongkrekic acid, a new inhibitor of mitochondrial ADP-ATP transport: radioactive labeling and chemical and biological properties; Lauquin GJ et al.; An isomer of bongkrekic acid, designated as isobongkrekic acid, has been isolated from ethereal extracts of Pseudomonas cocovenenans grown on defatted coconut . Isobongkrekic acid was also obtained by alkaline treatment of bongkrekic acid . Isobongkrekic acid possesses the same ultraviolet spectrum and the same molecular weight as bongkrekic acid; it has a similar infrared spectrum but not the same nuclear magnetic resonance (NMR) spectrum . The differences in NMR data were interpreted to mean that isobongkrekic acid differs from bongkrekic acid by the configuration of the dicarboxylic end; whereas the two carboxylic groups of the dicarboxylic end have the trans configuration in bongkrekic acid, they have the cis configuration in isobongkrekic acid . Differences between bongkrekic and isobongkrekic acids are lost after catalytic hydrogenation of the molecules . Isobongkrekic acid, like bongkrekic acid, is an uncompetitive inhibitor of ADP transport in mitochondria, provided the mitochondria are preincubated in the presence of the inhibitor and a minute concentration of ADP . The inhibitory and binding efficiency of isobongkrekic acid is considerably increased below pH 7 . The number of high affinity sites for {3H} isobongkrekic acid is 0.13 to 0.20 nmol/mg protein in rat liver mitochondria and about 1 nmol/mg protein in rat heart mitochondria, i.e., similar to the number of high affinity sites for {3H} bongkrekic acid . Isobongkrekic and bongkrekic acids compete for the same site, but the affinity of isobongkrekic acid for mitochondria is one-half to one-fourth that of bongkrekic acid.

Surgery, 1976 Jun, 79(6), 690 - 6
Evaluation of a prototype therapeutic system for prolonged, continuous topical delivery of homosulfanilamide in the management of Pseudomonas burn wound sepsis; Vistnes LM et al.; The ability of homosulfanilimide (HS) delivered from two different dressing vehicles to limit bacterial proliferation was evaluated in burned animals deliberately infected with virulent Pseudomonas organisms . Treatment consisted of once daily topical application of one of two vehicles: (1) an experimental prototype system that utilized micronized HS in a hydrophobic, bioerodible, polymeric matrix, impregnated on a fabric backing; or (2) a commercially available dressing that contained the same mass of drug in a hydrophilic cream base impregnated on the same backing . Wounds on control animals were covered with fabric backing with or without the bioerodible matrix . The experimental system was designed to maintain a finite local concentration of HS on the burn wound for at least 24 hours . It is known that the cream base presents a rapidly decreasing concentration of drug to the burn surface . HS delivered from the experimental system produced a significant reduction in deaths compared with the HS delivered from the cream base . In addition, the new method of delivering HS provided better control of local and systemic infection, and better wound hydration and also promoted earlier eschar separation . The experimental system was at least as convenient to apply as the cream and had an advantage with respect to inspection of the wound, since it could be removed and reapplied easily.

J Gen Microbiol, 1976 Jun, 94(2), 323 - 32
Influence of dilution rate on enzymes of intermediary metabolism in two freshwater bacteria grown in continuous culture; Matin A et al.; Two freshwater bacteria, a Pseudomonas sp . and a Spirillum sp., were grown in continuous culture under steady-state conditions in L-lactate-, succinate-, ammonium- or phosphate-limited media . In Pseudomonas sp., NAD-independent and NAD-dependent L-lactate dehydrogenases, aconitase, isocitrate dehydrogenase and glucose 6-phosphate dehydrogenase activities increased up to 10-fold as the dilution rate (D) was decreased from 0.5 to 0.02 h-1, regardless of whether the growth-limiting nutrient was carbon, ammonium or phosphate . In contrast, 2-oxoglutarate dehydrogenase and succinate dehydrogenase activities were not influenced by D, and NADH oxidase activity increased with D . Spirillum sp . gave different results in some respects, but it also exhibited an increase in the activity of several enzymes at low D values . Such increases may emanate from release of catabolite repression, and catabolite repressors for the five enzymes in Pseudomonas sp . showing such increases are probably compounds of carbon, nitrogen and phosphorus . It is likely that increased enzyme syntheses in low D cultures represent the normal physiological state for bacteria in aquatic environments where growth occurs slowly under nutrient limitations . Such increases probably permit a more effective utilization of nutrients present at sub-saturating concentrations.

Arch Microbiol, 1976 Jun, 108(2), 217 - 9
Oxygen metabolism in Pseudomonas methanica; Ferenci T; Three sites of oxygen metabolism in Pseudomonas methanica have been identified on the basis of studies of methane, ethanol and formate oxidation . The three oxidations exhibit different affinities for oxygen and sensitivities to inhibition by cyanide . The results obtained are consistent with the presence of a methane oxygenase and at least two terminal oxidases in Pseudomonas methanica.

J Gen Microbiol, 1976 Jun, 94(2), 400 - 4
The effects of proteins on bacterial attachment to polystyrene; Fletcher M; Bovine serum albumin, gelatin, fibrinogen and pepsin impaired the attachment of a marine pseudomonad to polystyrene Petri dishes, apparently through adsorption on the dish surface . Serum albumin also appeared to affect the bacterial surface . The basic proteins protamine and histone did not markedly inhibit attachment . These findings are discussed in relation to comparative experiments using tissue cells.

J Gen Microbiol, 1976 Jun, 94(2), 333 - 41
Influence of dilution rate on NAD(P) and NAD(P)H concentrations and ratios in a Pseudomonas sp . grown in continuous culture; Matin A et al.; A freshwater Pseudomonas sp . was grown in continuous culture under steady-state conditions in L-lactate-, succinate-, glucose- or ammonium-limited media . Under carbon limitation, the NAD(H) (i.e . NAD + NADH) concentration of the organisms increased exponentially from approximately 2 to 7 mumol/g dry wt as the culture dilution rate (D) was decreased from 0.5 to 0.02 h-1 . Organisms grown at a given D in any of the carbon-limited media possessed very similar levels of NAD(H) . Therefore, under these conditions, cellular NAD(H) was only a function of the culture O and was independent of the nature of the culture carbon source . D had no influence on the NAD(H) content of cells grown under ammonium limitation . In contrast, cellular NADH concentration was not influenced by D in carbon- or ammonium-limited media . In L-lactate-limited medium, bacteria possessed 0.14 mumol NADH/g dry wt; very similar levels were found in organisms grown in the other media . The results are consistent with those of Wimpenny & Firth (1972) that bacteria rigidly maintain a constant NADH level rather than a constant constant NADH: NAD ratio . NADP(H) (i.e . NADP + NADPH) and NADPH levels were also not influenced by changes in the culture carbon source or in D; in L-lactate-limited medium these concentrations were 0.97 and 0.53 mumol/g cell dry wt, respectively . The NADPH:NADP(H) ratio was much higher than the NADH:NAD(H) ratio, averaging 55% in carbon-limited cells.

Appl Environ Microbiol, 1976 Jun, 31(6), 875 - 9
Stabilization of a psychrotrophic Pseudomonas protease by calcium against thermal inactivation in milk at ultrahigh temperature; Barach JT et al.; The heat-stable extracellular protease of Pseudomonas sp . (isolate MC60) was investigated . Heat resistance of the enzyme in milk at sterilization temperature was dependent on the presence of Ca2+ . The half-life of the enzyme at ultrahigh temperature (149 C) in skim milk or milk-salts buffer with Ca2+ was approximately 7.0 s . Treatment of milk with chelators completely removed the heatstabilizing effect of milk . The enzyme was partially purified by ammonium sulfate precipitation and column chromatography on Sephadex G-100 . At 21 C the enzyme retained greater than 85% activity after exposure to pH values between 5 and 10 . Enzyme activity was reduced by metal chelating agents . Both Ca2+ and Zn2+ were required for optimal enzyme activity . Molecular weight was estimated at 48,000 by gel filtration.

Biochim Biophys Acta, 1976 May 13, 429(3), 817 - 27
Purification and characterization of butyrylcholine-hydrolyzing enzyme from Pseudomonas polycolor; Nagasawa T et al.; A butyrylcholine-hydrolyzing enzyme (EC 3.1.1.-) fo Pseudomonas polycolor IFO 3918 was purified approximately 9270-fold with a recovery of 9.9% by use of chromatographic techniques . The enzyme preparation appeared to be homogeneous when subjected to electrophoretic and ultracentrifugational analyses . The molecular weight was determined as approximately 59000 by gel filtration . Isoelectric focusing electrophoresis revealed that the enzyme had an isoelectric point around pH 5.1 . The enzyme catalyzed the hydrolysis of butyrylcholine with the miximum activity among various esters tested, and split benzoylcholine, propionylcholine and some aliphatic esters, but did not attact acetylcholine . The estimated value of Km at pH 7.5 and 25 degrees C was 7-10(-4) M for butyrylcholine . The enzyme was irreversibly inhibited by organophosphorus compounds and carbamates, such as diisopropylphosphofluoridate and eserine . The enzyme was inhibited by some compounds, such as atropine and quinidine . Auaternary ammonium salts showed an inhibitory effect on the enzyme resembling co-operative inhibition.

J Biol Chem, 1976 May 10, 251(9), 2754 - 60
The reduced nicotinamide adenine dinucleotide-activated phosphoenolpyruvate carboxylase from Pseudomonas MA . Correlation of allosteric properties with changes in the sedimentation behavior; Millay RH Jr et al.; Phosphoenolpyruvate carboxylase from Pseudomonas MA, grown on methylamine as a sole carbon source, has been studied with respect to some of its regulatory properties . The enzyme shows both negative and positive cooperativity with respect to the substrate phosphoenolpyruvate (Hill coefficients of 0.5 and 1.75) . The enzyme requires a divalent cation for activity . Either magnesium or manganous ion is effective . While magnesium shows normal kinetics, manganous ion shows positive cooperativity with a Hill coefficient of 1.4 . The enzyme is activated 50-fold by 0.2 mM NADH at 1 mM phosphoenolpyruvate . This activation is hysteretic, showing a lag of 2 to 3 min . Both NADH and Mn2+ induce a change in the sedimentation coefficient of the enzyme from 12.4 to 8.5 as measured by sucrose density gradient centrifugation . High concentrations of phosphate or sulfate are capable of producing this effect on sedimentation, but neither will activate more than 3-fold . Thus, if NADH is an indicator of the total energy level of the cell, the enzyme appears to be susceptible to control by factors which reflect this total energy level . The importance of this control with respect to hypothetical pathways of carbon utilization in the organism is discussed.

J Biochem (Tokyo), 1976 May, 79(5), 937 - 43
Purification and properties of a four iron-four sulfur protein from a Pseudomonas species; Matsumoto T et al.; We have isolated an iron-sulfur proteins from a Pseudomonas species grown on glucose . This protein has different properties from the two known iron-sulfur proteins isolated from other Pseudomonas species: rubredoxin and putidaredoxin . The iron-sulfur protein was purified to homogeneity by DEAE-cellulose column chromatography and Sephadex G-75 gel filtration . The absorption spectrum of the oxidized iron-sulfur protein shows a peak at 283 nm with shoulders at about 290, 320, and 410 nm . The protein contains 4 g atoms of iron and 4 moles of labile sulfur per mole of protein, and has a molecular weight of approximately 14,000 . The amino acid composition of the protein shows a predominance of acidic amino acids . The Pseudomonas protein was found to be active for both photosynthetic nicotinamide nucleotide reduction by chloroplasts and cytochrome c reduction by spinach ferredoxin-NADP+ reductase {EC 1.6.7.1} . On the basis of these results, this protein appears to be unique among all known ferredoxins . From an evolutionary point of view, it appears to be more closely related to Azotobacter ferredoxin than to Desulfovibrio ferredoxin.

J Virol, 1976 May, 18(2), 701 - 8
Localization and functional role of the pseudomonas bacteriophage 2 depolymerase; Castillo FJ et al.; The adsorption apparatus of phage 2 consits of a symmetrical base plate of snowflake appearance, composed of six droplike spikes 7.0 to 7.5 nm in length with a maximum diameter of 4.5 to 5.0 nm . The spikes are attached by their narrow ends to a central ring 7.0 to 7.5 nm in diameter . Phage 2 deopolymerase, a phage 2-induced hydrolytic enzyme, was found to be a structural protein of phage 2 or in close association with the base plate . Pdp1, a phage 2 mutant, possesses a polypeptide that is antigenically similar to the depolymerase, but devoid of hydrolytic activity . This polypeptide was found to be located in the region of the base plate of pdp1 . Treatment of intact cells of strain BI with purified phage 2 depolymerase inhibited the adsorption of phage 2 . When phage receptor-containing fractions of slime glycolipoprotein and lipopolysaccharide were hydrolyzed by the depolymerase, amino sugars were released, and the phage-inactivating activities of these fractions were lost . The depolymerase was also observed to induce the lysis of strain BI cells in hypotenic medium . The phage 2 depolymerase appears to play a role in adsorption and release of phage.

J Clin Chem Clin Biochem, 1976 May, 14(5), 225 - 6
A new enzymatic method for the determination of free and conjugated glucuronic acid; Wagner G et al.; A new method is reported for the quantitative determination of glucuronic and galacturonic acid, which is based on spectrophotometric measurement of NADH . The NAD-linked oxidation of the uronic acids to the corresponding dicarboxylic acids is measured in the presence of uronic acid dehydrogenase . This enzyme was isolated from Pseudomonas syringae . The test is highly specific for glucuronic and galacturonic acid and permits the exact determination of free and conjugated glucuronic acid . This enzymatic determination of glucuronic is the most sensitive method available today.

Nouv Presse Med, 1976 May 1, 5(18), 1193 - 5
{The use of new immunoglobulin preparation, enriched in IgA and IgM (IgGAM)}; Blatrix C et al.; The clinical results obtained with fraction IgGAM are reported . Different types of antibody deficiency syndromes have successfully been treated : 8 cases of Bruton-type agammaglobulinemia . In one of these case a tenacious Pseudomonas infection cleared off during the treatment . Two cases of non sex-linked familial agammaglobulinemia . Three cases of isolated IgM deficiency . Five cases of isolated IgA deficiency . Five cases with Soothill type IgA deficiency associated with high IgE levels . Five cases of septicemia of the new-born . Three cases with acquired agammaglobulinemia and in the premature infant (5 cases) . No side-effects nor appearance of anti-IgA antibodies have been observed.

Biokhimiia, 1976 May, 41(5), 864 - 8
{Study of the initial reaction of enzymatic oxidation of 1,8-dimethylnaphthalene}; Tsfasman IM et al.; Crude enzymatic preparation has been obtained from Pseudomonas bacteria which oxidises 1,8-DMN during 10-hour incubation with the following formation of the same products which are formed when this compound is oxidized by the intact cells . The first product of the oxidation is 1-methyl-8-oxymethylnaphtalene (compound I), obtained as a result of hydroxylation of one methyl group . Probably hydroxylase of 1,8-DMN may be referred to the class of oxigenases of the basis of the absence of 18O incorporation from H218O to compound I, and also resulting from the data on absorption of molecular oxygen during the reaction . The enzyme is completely inhibited by chelating agents of Fe2+ NAD(P)H and Fe2+ stimulates the reaction of 1.8 DMN oxidation.

Appl Environ Microbiol, 1976 Apr, 31(4), 551 - 61
Colonization of soil by Arthrobacter and Pseudomonas under varying conditions of water and nutrient availability as studied by plate counts and transmission electron microscopy; Labeda DP et al.; Arthrobacter globiformis and a Pseudomonas soil isolate were incubated separately and in combination in soil that had been presterilized by autoclaving . Growth and other responses of the cells in situ in this soil were monitored by plate counts and transmission electron microscopy examinations of cell sections . During the soil incubations, some of the samples were first allowed to dry and then were remoistened with water or with a dilute or a concentrated nutrient solution . Based on plate counts and ultrastructural analysis . Arthrobacter seemed to be in a non-multiplying coccoid-rod resting state and to be virtually immune to soil drying . Addition of a dilute nutrient solution helped maintain cell ultrastructure and prevent a low level of lysing that occurred in the absence of nutrient addition . Addition of a concentrated nutrient solution brought on cell multiplication as both coccoid-rods and long rods, but the ultimate form with further incubation was the coccoid-rod . The Pseudomonas strain suffered death and ultrastructural deterioration as water became less available . It responded by cell multiplication to an equal extent when either water or dilute nutrients were added, but possibly was able to give a growth response to nutritive amendment when a concentrated nutrient addition was made . The Arthrobacter was not affected by the presence of Pseudomonas in dual culture . The Pseudomonas, however, possibly suffered a nutritive deficiency under these conditions.

Biochim Biophys Acta, 1976 Apr 8, 429(2), 616 - 23
Substrate specificity of the L-serine O-sulphate degrading activities of Pseudomonas FR; Tudball N et al.; 1 . Two enzyme systems obtained from Pseudomonas FR capable of catalysing the alphabeta-elimination of L-serine O-sulphate exhibit a wide range of substrate specificity . Greatest activity was exhibited towards beta-substituted serine and cysteine derivatives . Enzyme A shows a marked preference for the L-isomeric form and enzyme B shows a preference for D-isomers . 2 . The alternative activities were shown to be properties of the same enzyme by inhibition properties and heat denaturation experiments . 3 . The assay of enzyme A by a number of alternative substrates at various stages during its purification confirmed the multi-substrate specificity of the system . 4 . Growth of Pseudomonas FR on S-methyl-L-cysteine as the sole carbon source also resulted in the induction of enzyme B . Growth patterns and levels of induced enzyme were similar to those obtained when L-serine O-sulphate was employed in comparable circumstances.

Aust N Z J Med, 1976 Apr, 6(2), 156 - 7
Meliodosis presenting as encephalitis; Singh N; Meliodosis is an infectious disease caused by Pseudomonas pseudomellei, an organism that is common in South-East Asia, the Caribbean and northern Australia . In the author's case report the disease presented as an encephalitic illness with fever and epileptic fits in a five-year-old Chinese boy . The case illustrates one of the many forms of this illness . The patient, though very ill, made an excellent recovery . The report discusses the manifestations, diagnosis and management of Meliodosis.

J Gen Microbiol, 1976 Apr, 93(2), 259 - 65
A biochemical basis for obligate methylotrophy: properties of a mutant of Pseudomonas AM1 lacking 2-oxoglutarate dehydrogenase; Taylor IJ et al.; Pseudomonas AMI is a facultative methylotroph which grows on a wide range of carbon compounds . A mutant of Pseudomonas AMI (ICT4I) grew only on C1 compounds and is thus an artificial obligate methylotroph . Measurements of activities of the components of the 2-oxoglutarate dehydrogenase complex suggest that the E2 component (dihydrolipoamide transsuccinylase) is not functional . All other tricarboxylic acid cycle enzymes were present with activities comparable to those in wild-type Pseudomonas AMI and cytochrome levels were unchanged in the mutant . Suspensions of the mutant oxidized pyruvate, lactate, beta-hydroxybutyrate, acetoacetate and 2-oxoglutarate at very low rates . By contrast, C1 compounds were oxidized at the same rate as in wild-type bacteria . Two revertants of ICT4I which regained 2-oxyoglutarate dehydrogenase activity also regained the ability to oxidize and grow on the same substrates as wild-type bacteria . It is concluded that lack of 2-oxoglutarate dehydrogenase may well be the basis of obligate methylotrophy in some bacteria.

Antibiotiki, 1976 Apr, 21(4), 344 - 9
{Relationship between beta-lactamase activity of clinical strains of Pseudomonas pyocyanea and their resistance to penicillins}; Gromova GN; The study of 50 clinical strains of Ps . aeruginosa revealed their high resistance to benzylpenicillin, ampicillin, oxacillin, streptomycin and kanamycin with a tendency to polyresistance . The same strains were highly sensitive to gentamycin and polymyxin . The strains produced constitutive beta-lactamase in small amounts . The enzyme synthesis in Ps . aeruginosa was induced by benzylpenicillin . Still high concentrations of it were required for the induction . The maximum induction of beta-lactamase synthesis was observed by the 6th hour of the inductor addition . The maximum induction level was observed 4 hours after addition of benzylpenicillin . The level of induction of beta-lactamase synthesis in the strains ranged within 5-93 . A significant part of the enzyme was liberated from the cells during induction . Interaction between the induction level of beta-lactamase synthesis and resistance of Ps . aeruginosa to penicillins was found.

Arch Otolaryngol, 1976 Apr, 102(4), 236 - 7
Malignant external otitis in children; Joachims HZ; Malignant external otitis is a very serious variant of external ear infection . It is caused by Pseudomonas aeruglinosa, affects elderly diabetic patients and bears high mortality . The disease may occur in children in whom malnutrition and anemia create suitable conditions for P aeruginosa infection . The disease differs in children, it invades the middle ear and facial nerve in its intratemporal course . The treatment should consist of large doses of carbenicillin sodium and gentamicin sulfate, together with the thorough surgical debridement of all infected tissues.

Eur J Biochem, 1976 Apr 1, 63(2), 427 - 9
Horse-liver alcohol dehydrogenase and Pseudomonas testosteroni 3(17)beta-hydroxysteroid dehydrogenase transfer epimeric hydrogens from NADH to 17beta-hydroxy-5alpha-androstan-3-one . An exception to one of the Alworth-Bentley rules; Groman EV et al.; In the reduction of 17beta-hydroxy-5alpha-androstan-3-one to the 3beta-alcohol, horse liver alcohol dehydrogenase utilizes the 4-pro-R hydrogen of NADH whereas the 3(17)beta-hydroxysteroid dehydrogenase of Pseudomonas testosteroni utulized the 4-pro-S hydrogen . These observations provide an exception to the rule proposed by Alworth and Bentley that with regard to the paired methylene hydrogens at C-4 of NADH and NADPH "the stereospecificity of a particular reaction is fixed and does not vary with the source of the enzyme preparation" . It is also apparent that for these two enzymes, the selection of the side of NADH from which hydride is transferred to substrate cannot in both cases be dictated by the "best fit" of substrate and cofactor.

Arch Microbiol, 1976 Mar 19, 107(2), 225 - 7
Isolation of regulatory mutants of Pseudomonas acidovorans by use of amino acid analogs; Thevenet NJ et al.; Mutants resistance to various combinations of threonine, lysine and/or their analogs were obtained and characterized in Pseudomonas acidovorans . In particular, mutants resistant to aminoethylcysteine had a dihydrodipicolinate synthetase insensitive to lysine inhibition whereas mutants resistant to threonine plus a low concentration of aminoethylcysteine had a feedback-insensitive aspartokinase.

Eur J Biochem, 1976 Mar 16, 63(1), 233 - 40
Purification and properties of a methanol-oxidizing enzyme in Pseudomonas C; Goldberg I; A methanol-oxidizing enzyme has been purified from Pseudomonas C, grown on methanol as a sole source for carbon and energy . The purification procedure involved ammonium sulphate precipitation, ion-exchange chromatography and gel filtration and resulted in a yield of 35.4% . Enzyme activity can be coupled to phenazine methosulfate and requires the presence of ammonium ions in the assay mixtures . The enzymes possesses a broad specificity for primary alcohols . Formaldehyde is also oxidized by the purified enzyme . The Km value for methanol is 15 muM . The optimum pH for the oxidation of both methanol and formaldehyde is about 10.4 . The enzyme has a molecular weight of about 128000 and consists of two subunits each having a molecular weight of 60000.

Eur J Biochem, 1976 Mar 16, 63(1), 199 - 209
Purification and properties of cyclopentanone oxygenase of Pseudomonas NCIB 9872; Griffin M et al.; 1 . Cyclopentanone oxygenase from Pseudomonas NCIB 9872 has been purified some 40-fold . It gives a single peak in the ultracentrifuge and a single major protein band on polyacrylamide gels contaminated with about 5% of a slower migrating impurity . Flavin dissociates from the protein during electrophoresis . 2 . The enzyme has a molecular weight of about 200000 and is a homopolymeric assemblage of either three of four subunits of molecular weight 54000-58000 . 3 . The prosthetic group is FAD and values of about 2.5 are typically obtained for the number of moles bound to each mole of holoenzyme . Some FAD probably dissociates during purification and it seems likely that each subunit binds one FAD in the undamaged protein.

J Clin Pathol, 1976 Mar, 29(3), 224 - 7
Chemotaxis, random mobility, and mobilization of polymorphonuclear leucocytes in malnutrition; Chandra RK et al.; Neutrophil mobilization following administration of Pseudomonas polysaccharide was significantly reduced in malnutrition, especially during infection . The random mobility of polymorphonuclear leucocytes (PMNs) was slightly decreased in undernutrition . Chemotactic migration of PMNs was depressed and correlated more with the presence of infection than with nutritional deficiency . It is possible that these abnormalities of PMN mobilization and mobility in malnourished individuals contribute to suboptimal amount, kinetics, and pattern of tissue inflammatory response to bacteraemic challenge.

J Bacteriol, 1976 Mar, 125(3), 910 - 5
Diminution of outer membrane permeability by Mg2+ in a marine pseudomonad; Moustafa Hassan H; Intact cells of the marine pseudomonad MB-45, in the presence of optimal Mg2+, exhibited little alkaline phosphatase activity as judged by the hydrolysis of p-nitrophenylphosphate . Sonic extracts, in contrast, were rich in this activity . Removal of the loosely bound outer layer did not diminish this crypticity of alkaline phosphatase, but decreasing the concentration of Mg2+ in the suspending medium progressively exposed the alkaline phosphatase . Since MB-45 did not liberate alkaline phosphatase into the surrounding medium even in the absence of Mg2+ and since this enzyme is localized in the periplasmic space, it can be concluded that the crypticity was due to the exclusion of p-nitrophenylphosphate by the outer membrane . Mg2+ is apparently essential for the full expression of this limited permeability.

Invest Ophthalmol, 1976 Mar, 15(3), 211 - 3
Deposits of mucosubstances on the cornea by topical chloramphenicol: an electron microscopic study; Mitsui Y et al.; Chloramphenicol was instilled into rabbit eyes and five days later the corneal surface was examined by scanning electron microscopy . The corneal surface was roughened and showed a proliferative appearance . Examination of thin sections of the cornea by transmission electron microscope showed that the corneal surface stained strongly with alcian blue . It was thus supposed to be a deposit of mucosubstances . Mucosubstances increase pathogenicity of bacteria and, therefore, this deposit may accelerate Pseudomonas keratitis after chloramphenicol application.

Can J Microbiol, 1976 Mar, 22(3), 404 - 8
The distribution of the isocitrate lyase serine pathway amongst one-carbon utilizing organisms; Bellion E et al.; A study of several one-carbon-utilizing organisms was conducted to determine the distribution of the recently found isocitrate-lyase-positive serine pathway of C1 assimilation . The results showed that this pathway is restricted to soil-isolated, non-pigmented Pseudomonas, initially isolated in methylamine enrichments, and to certain species of Hyphomicrobium . It was not detected in any organisms possessing a pink pigment.

Chest, 1976 Mar, 69(3), 338 - 44
Pressure transducers as a source of bacteremia after open heart surgery . Report of an outbreak and guidelines for prevention; Weinstein RA et al.; During four weeks in 1974, eight (26 percent) of 31 intensive care unit patients who had undergone open-heart surgery developed symptomatic Pseudomonas cepacia bacteremia in the intensive care unit one to three days after the open-heart surgery . An investigation demonstrated that operating room pressure transducers were being contaminated during cleaning with a detergent that contained P cepacia at the rate of 10(4) organisms per milliliter and that the organisms were transmitted to patients after open-heart surgery as a result of one to three days of contact with transducer-monitoring lines used in the operating room and brought to the intensive care unit with the patient . Pressure-transducer contamination, a frequently unappreciated but preventable cause of nosocomial bacteremia, can be minimized by sterilizing transducers between use on different patients by paying strict attention to aseptic technique when setting up, calibrating, and using monitoring systems; and by changing transducers, tubing, and monitoring fluid for each monitored patient at regular intervals.

Can J Microbiol, 1976 Mar, 22(3), 429 - 31
Soybean flower-to-seed movement of epiphytic bacteria; Leben C; Epiphytic tracer bacteris (a Pseudomonas sp . and an Arthrobacter sp.) from soybean buds were introduced into open flowers of greenhouse-grown soybean plants . Of the 177 resulting pods cultured after surface disinfection, tracers were recovered from within 24 . Seed from two of these pods also carried tracers.

Mikrobiologiia, 1976 Mar-Apr, 45(2), 310 - 2
{Alpha-ketoglutaric acid transamination of synthetic omega-amino acids in bacteria}; Naumova RP et al.; Bacteria utilizing synthetic lactams and omega-amino acids contain enzymes involved in transamination of epsilon-aminocaproic, zeta-aminoenanthic, eta-aminocaprilic acids with alpha-ketoglutaric acid . Natural and synthetic omega-amino acids are not inductors of the synthesis of corresponding transaminases . A strain of Pseudomonas dacunchae has been selected; it possesses an elevated activity of transamination of synthetic omega-amino acids, including zeta-aminoenanthic and eta-aminocaprilic acids.

Invest Ophthalmol, 1976 Mar, 15(3), 208 - 10
The role of mucin on experimental Pseudomonas keratitis in rabbits; Mitsui Y et al.; The role of mucin in the manifestation of Pseudomonas keratitis was studied . Pseudomonas was cultivated in solutions of mucin, in which it grew rapidly and then inoculated into rabbit cornea by needle pricks . When the organism was inoculated as a suspension in saline, infection infrequently occurred as small ring abscesses of short duration around a few sites of inoculation . When the organism was inoculated as a suspension in a solution of gastric mucin, infection was usually observed as severe hypopyon-keratitis with formation of a huge ring abscess . Corneal perforation and panophthalmitis resulted in some cases . It was thus concluded that the pathogenicity of Pseudomonas is definitely increased when it was inoculated into the cornea with mucin solution.

Eur J Biochem, 1976 Feb 16, 62(2), 257 - 62
Phase transitions in the membrane of a marine bacterium, Pseudomonas BAL-31; Tsukagoshi N et al.; An unsaturated fatty acid auxotroph, strain UFA, isolated from the marine pseudomonad Pseudomonas BAL-31, host cell of the lipid-containing bacteriophage PM2, was grown in media supplemented with different unsaturated fatty acids . Under these conditions the fatty acid composition of the cell could be altered drastically . The phase transition in the native membrane and in the extracted lipids was analyzed by electron spin resonance using a nitroxide spin probe . Membranes prepared from strain UFA grown in cis16:1 or trans16:1 showed one transition at 9.4 degrees C and 12.4 degrees C respectively . Extracted lipids in both cases had almost the same transition temperature as that of the intact membrane . Membranes prepared from Pseudomonas BAL-31 had one transition at approximately 12 degrees C, on the other hand there was no clear cut phase transition using extracted lipids . Replication of bacteriophage PM2 took place below the transition temperature of the membrane lipids in the case where strain UFA was grown in tran16:1 . Other cases were not studied.

Biochim Biophys Acta, 1976 Feb 13, 422(2), 280 - 94
Purification, crystallization and properties of iron-containing superoxide dismutase from Pseudomonas ovalis; Yamakura F; Three electrophoretically distinct superoxide dismutases (EC 1.15.1.1) were observed in the crude extracts from Pseudomonas ovalis . One of these was isolated as an iron-containing superoxide dismutase . It contained 1.4 gatoms of Fe per mol of enzyme, and had a specific activity of 3900 units per mg of protein . A crystallized enzyme contained 1.1 gatoms of Fe per mol of enzyme, and had a specific activity of 3100 units per mg of protein . The results of sedimentation equilibrium and gel filtration indicated a molecular weight of 40,000 . S020,W was estimated as 3.18 by sedimentation velocity study . Sodium dodecyl sulfate gel electrophoresis indicated that the enzyme was composed of two subunits, and had a molecular weight of 19,500 . Analysis for sulfhydryl groups showed that there were four such groups per mol of enzyme . The spectrum of visible and ultraviolet region, the amino acid composition, the CD spectrum of the enzyme, and the effect of certain compounds on the enzyme, were studied and compared with iron-containing superoxide dismutases isolated from other organisms.

Can J Microbiol, 1976 Feb, 22(2), 154 - 8
Temporal origin of viral phospholipids of the enveloped bacteriophage phi 6; Sands JA et al.; The enveloped bacteriophage phi 6 contains a higher relative level of the negatively charged phospholipid phosphatidylglycerol than is found in the membranes of the host bacterium . During infection of Pseudomonas phaseolicola with phi 6, the level of phosphatidylglycerol synthesis increases significantly . The lipid used to form the viral envelope consists almost entirely of cellular phospholipids synthesized before infection and phosphatidylglycerol synthesized after infection . Based on these and previously published results, a speculative model for this viral envelope formation process is presented.

Pediatrics, 1976 Feb, 57(2), 239 - 43
Pseudomonas cepacia in an intensive care nursery; Rapkin RH; A small epidemic of Pseudomonas cepacia infection in an intensive care nursery was associated with contaminated, distilled water . This appeared to have occurred because of a leak in the distiller, contaminating its effluent . The distilled water was not subsequently sterilized, nor were the bottles used to distribute it . The epidemic was promptly terminated by substitution of sterile, distilled water distributed in sterile containers . Both professional and nonprofessional hospital personnel failed to oversee the appropriateness of handling of patient care materials.

J Bacteriol, 1976 Feb, 125(2), 679 - 88
Structure of plain and complex flagellar hooks of Pseudomonas rhodos; Raska I et al.; The proximal hooks of plain and complex flagella produced by a strain of Pseudomonas rhodos have been analyzed by electron microscopy and optical diffraction and filtering . Plain flagellar hooks are cone-shaped, 70 nm long, and 13 to 21.5 nm wide, and consist of helically arranged subunits . Complex flagellar hooks are cylinders, 180 to 190 nm long, and 15 to 16 nm wide, and are composed of globular subunits . The structure comprises four small-scale helical rows of subunits intersecting bewteen 10 and 11 large-scale helices of pitch angle 80 degrees . The axial and lateral dimensions of the unit cell, which define the surface lattice, are 4.9 and 4.7 nm, respectively . In addition, a core structure, approximately 5 nm wide, has been demonstrated inside the hook cylinder . Complex flagellar hooks were isolated and purified by gradient centrifugation after acid degradation of the attached filaments . Isolated hook particles have an average sedimentation constant of 130S and consist of a protein of molecular weight 43,000 . A model of the complex flagellar hook is presented, and its possible role in flagellar assembly and rotation is discussed.

Eur J Biochem, 1976 Jan 15, 61(2), 337 - 44
The reduction of plastocyanin by plastoquinol-1 in the presence of chloroplasts . A dark electron transfer reaction involving components between the two photosystems; Wood PM et al.; The reduction of plastocyanin by plastoquinol-1 was efficiently catalysed by disrupted chloroplasts or etioplasts in the dark . The reaction was inhibited by 2,5-dibromomethylisopropyl-p-benzo-quinone which inhibits photosynthetic electron transport between plastoquinone and cytochrome f . Evidence is presented that the reduction took place via cytochrome f, and that plastoquinone-9 was not involved . Triton X-100 and organic solvents were inhibitory, but partial fractionation was achieved without loss of activity by density gradient centrifugation in the presence of high digitonin concentrations . All active material contained cytochromes b-559LP and b-563 in addition to cytochrome f, but these b-type cytochromes were not directly involved . Other 1-electron acceptors could be used in place of plastocyanin, for instance ferricyanide and Pseudomonas cytochrome c-551 . The reaction can be applied to give a sensitive dark assay for active cytochrome f . It is suggested that cytochrome f possesses two sites for interaction with redox reagents: a hydrophilic site with which plastocyanin reacts by electron transfer and a hydrophobic site with which plastoquinol reacts by hydrogen atom transfer.

Eur J Biochem, 1976 Jan 15, 61(2), 589 - 96
Uronic acid dehydrogenase from Pseudomonas syringae . Purification and properties; Wagner G et al.; 1 . Uronic acid dehydrogenase was purified to homogeneity . After a 338-fold purification a yield of 16% was achieved with a specific activity of 81 mumol NADH formed min-1 mg protein-1 . 2 . The purity of the enzyme was controlled by disc electrophoresis, sodium dodecylsulfate electrophoresis and ultracentrifugation . 3 . A molecular weight of 60 000 was determined by gel chromatography and by ultracentrifugation . 4 . The native enzyme is composed of two subunits, their molecular weight being 30 000 as estimated by sodium dodecylsulfate electrophoresis . The subunits as such are inactive . 5 . The absorption spectrum with a maximum at 278 nm shows no evidence for a prosthetic group . 6 . For catalytic activity no SH groups and no metals seem to be necessary . 7 . The Michaelis constants determined with the pure enzyme are for glucuronic acid Km = 0.37 mM, galacturonic acid Km = 54 muM and NAD+ (with glucuronic acid) Km = 80 muM . 8 . A weak reverse reaction could be observed with glucaric acid lactones at acidic pH . 9 . NADH is competitive with NAD+ . The inhibitor constant is Ki = 60 muM . 10 . The NAD+ binding site seems to be of lower specificity than the uronic acid binding site.

Lancet, 1976 Jan 3, 1(7949), 31 - 3
Melioidosis antibodies in Commonwealth soldiers; Thin RN; Titres of melioidosis haemagglutinating antibodies of 1/40 or more were found in 18 of 905 British, Australian, and New Zealand soldiers serving in West Malaysia . Previous mild unsuspected melioidosis seemed to be responsible for these positive titres, which were more common in men exposed to surface water at work and during recreation . This accords with the current view that soil and surface water is the normal habitat of Pseudomonas pseudomallei, the causal organism . Pyrexia of unknown origin after arriving in Malaysia was significantly more common in men with titres of 1/40 or more than in the remainder . It is suggested that mild melioidosis may present as pyrexia of unknown origin . Pyrexias of unknown origin should be investigated vigorously in patients who are in or who have visited endemic areas.

J Clin Microbiol, 1976 Jan, 3(1), 62 - 6
Pseudomonas cepacia strains isolated from water reservoirs of unheated nebulizers; Gelbart SM et al.; Pseudomonas cepacia strains were isolated from the water reservoirs of unheated nebulizers in a hospital setting . The isolates were characterized by morphology, biochemical tests, and antibiotic susceptibility patterns . An interesting feature of these organisms was their capacity for sustained multiplication in either doubly deionized or doubly distilled water as well as 5% dextrose or 0.9% saline . They could not multiply in doubly deionized and then doubly distilled water or in any of several parenteral nutrition solutions studied . Isolation of Pseudomonas cepacia from the water reservoirs of unheated nebulizers suggests that this equipment may serve as a source of respiratory tract exposure to the organism.

Trans R Soc Trop Med Hyg, 1976, 70(4), 335 - 7
A retrospective serological suvey of Royal Marines previously exposed to Pseudomonas pseudomallei in South East Asia; Preston PJ et al.; Following the suggestion that it was possible that cases of melioidosis amongst those who had been exposed abroad in the past, might be escaping notice, 487 Royal Marines were examined by indirect haemagglutination studies . Four hundred and eleven of these subjects had served for variable times in areas where melioidosis has been known to occur in Indonesia and Malaya, between 1960 and 1974, occupied in activities in the jungle and paddy fields during which exposure to the disease was to be expected . No evidence of residual subclinical melioidosis was found and it seems unlikely that recrudescent disease will prove to be a problem in the future for English servicemen who have been in South East Asia.

J Supramol Struct, 1976, 5(1), 73 - 9
Control of membrane morphogenesis in bacteriophage PM2; Brewer GJ; The regulation of membrane formation in bacteriophage PM2 serves as a simple model for changes in membrane structure in eukaryotic cells . Prior to Pseudomonas host lysis, wild-type virions mature to an icosahedral morphology at the inner face of the cytoplasmic membrane . The preliminary characterization of two temperature-sensitive mutants of PM2 is described . In cells infected at the restrictive temperature with ts 1, an abundance of "empty" virus-size membrane vesicles are seen . Synthesis of DNA is also reduced in ts 1 infected cells . The preponderance of vesicles is not seen in cells infected with wild-type virus or with ts 1 at the permissive temperature . The "empty" appearance of the viral membranes suggests that viral DNA is not encapsulated . The major viral capsid protein (MW 26,000) is located just outside the viral membrane and normally sidiments with host and virus membranes . This protein made by mutant ts 5 does not pellet with these membranes; instead, large amounts of capsid protein can be precipitated from the supernatant with TCA . Compared to cells infected with wild type virus, cells infected with ts 5 at the restrictive temperature produced inside the cell an abundance of virus-size membrane vesicles . Taken together, these results with viral mutants suggest that formation of a viral membrane of the proper size does not require a DNA core around which to form, or an outer scaffolding of coat protein against which to form a spherical bilayer.

Appl Environ Microbiol, 1976 Jan, 31(1), 78 - 82
Enzymatic epoxidation: synthesis of 7,8-epoxy-1-octene, 1,2-7,8-diepoxyoctane, and 1,2-Epoxyoctane by Pseudomonas oleovorans; Schwartz RD et al.; The kinetics of the enzymatic formation of 7,8-epoxy-1-octene, 1,2-7,8-diepoxyoctane, and 1,2-epoxyoctane by growing and resting cell suspensions of Pseudomonas oleovorans are described . Formation of 1,2-epoxyoctane occurs concurrently with exponential growth on 1-octene, providing that 1-octene is in excess . Conversion of 1,7-octadiene to 7,8-epoxy-1-octene by cells growing on octane lags behind exponential growth and continues into the stationary phase, terminating upon cell death . Formation of 1,2-7,8-diepoxyoctane does not begin until the cells are well into the stationary phase and also continues until cell death . Results with growing and resting cell suspensions suggest that the various substrates compete for the same enzyme system; that viable cells are essential for substrate transport and epoxidation by whole cells; and that whole cells may concentrate and sequester the epoxides, rendering them unrecoverable by our current methods.

Acta Biochim Pol, 1976, 23(4), 375 - 86
Oxidation of methanol by facultative and obligate methylotrophs; Michalik J et al.; 1 . The newly isolated methanol obligate Methylomonas sp . and the methanol facultative Pseudomonas sp . oxidize methanol at an unchanged rate over concentration range from 0.1 to 600 mM; the oxidation rate by the obligate methylotroph is 2.5 times higher (300 nmoles O2/min/mg dry wt.) . Low-molecular alcohols, formaldehyde and formate serve as respiratory substrates for the intact cells of both methylotrophs . 2 . Methanol dehydrogenase of both methylotrophs isolated should be classified as the phenazine methosulphate-dependent pteridine-type enzyme of double methanol-and formaldehyde-dehydrogenase function . This soluble enzyme is stimulated about 10-fold by NH+4, which results in enhancement of V max, and shows the same specificity and the same affinity toward methanol and formaldehyde (K m about 5 X 10(-5) M) . Heat-inactivation of the 10-fold purified enzyme is associated with the release of a watersoluble pigment with maximum fluorescence at 420-430 nm . 3 . NAD-deendent formate dehydrogenase was found to catalyse the third step of methanol oxidation in both methylotrophs.

Mikrobiologiia, 1976 Jan-Feb, 45(1), 23 - 7
{Formation of L-asparagine and L-glutamine deamidases by bacterial cultures}; Vinogradov BD et al.; Among studied 40 bacterial cultures, 17 strains catalysed hydroxylaminolysis of I.-asparagine and L-glutamine, and among these cultures seven strains belonged to the Pseudomonas genus . Extracts of the cells of Ps . boreopolis 526 (MGU), Ps . aurantiaca IBFM B-14, and Ps . septica IBFM B-40 had the maximum deamidase activity during the stage of decelerated growth . The activity of L-asparaginase 2 was practically the same upon various ways of disintegration of the cells of Ps . aurantiaca IBFM B-14: ballistic technique, ultrasound, extrusion form the solid state . Production of beta-aspartylhydroxamic acid by L-asparaginase 2 depended on the concentration of hydroxylamine and protein in the reaction mixture and on the duration of the reaction.

Acta Chem Scand B, 1976, 30(8), 721 - 6
Pseudomonas cytochrome c peroxidase . XII . Product inhibition studies; Ronnberg M; Kinetic studies of the reaction mechanism of Pseudomonas cytochrome c peroxidase (PaCCP) were made by the method of product inhibition using oxidized cytochrome C (551 P.aeruginosa) and oxidized Pseudomonas azurin as products . Inhibition by the two oxidized substrates was linearly non-competitive towards the respective reduced electron donor and towards hydrogen peroxide . Although a full kinetic analysis is experimentally impossible in a peroxidase-type reaction, the results do provide some evidence in favour of an ordered reaction mechanism in which hydrogen peroxide is the first to add to PaCCP and electron donor the second.

Z Naturforsch {C}, 1976 Jan-Feb, 31(1-2), 91 - 7
{Chromosomal structures of Pseudomonas testosteroni . I . Isolation and characterization of the chromosomal complexes . (author's transl)}; Reimer G et al.; After lysis of Pseydomonas testosteroni with lysozyme and non-ionic detergents different DNA-protein complexes can be separated in 5-25% (w/v) neutral sucrose gradient . The protein to DNA ratio of these complexes varies between 0.5-4.5 to 1, whereby the faster sedimenting forms contain more protein than the slower sedimenting ones . Different initial rates of DNase digestion may indicate various degrees of DNA packing in these complexes . The chromosomal complexes of Pseudomonas testosteroni are relatively stable towards pronase . Treatment with RNase or sodium dodecylsulphate is accompanied by a dramatic increase in viscosity and decrease in relative density . It suggests that DNA in these complexes is maintained in its supercoiled form by RNA molecule(s) in a similar way as in isolated chromosome of E . coli.

Antonie Van Leeuwenhoek, 1976, 42(1-2), 145 - 55
Autolysis of high-GC isolates of Pseudomonas putrefaciens; Levin RE et al.; High-GC isolates of P . putrefaciens undergo extensive autolysis after growth, resulting in a marked decrease in turbidity and the release of high-molecular-weight DNA which imparts a high viscosity to culture broths . The native DNA released is resistant to attack by the exocellular DNase activity of the culture broths . Autolysis is inhibited by a pH of 6.0 and the presence of 0.001 M Mg++ or Ca++, and is enhanced by elevated pH values and temperatures . This autolytic phenomenon in broth cultures readily distinguishes high- from low-GC isolates . The latter do not exhibit autolysis.

Biochim Biophys Acta, 1975 Dec 18, 410(2), 333 - 46
Purification and some properties of a novel maltohexaose-producing exo-amylase from Aerobacter aerogenes; Kainuma K et al.; Maltohexaose producing amylase (EC 3.2.1.-) is the fourth known exo-amylase, the three previously known being glucoamylase, beta-amylase and Pseudomonas stutzeri maltotetraose producing amylase . The enzyme after release from Aerobacter aerogenes cells by 0.1% sodium lauryl sulfate extraction was purified by ammonium sulfate precipitation, DEAE-Sephadex column chromatography and Sephadex G-100 gel filtration to 80-fold of the original sodium lauryl sulfate extract activity, It gave a single band on disc electrophoresis, and the molecular weight by gel filtration was 54 000 . This amylase showed maximal activity at 50 degrees C and pH 6.80 . The pH stability range was relatively wide, the enzyme retaining more than 90% of its initial activity in the range of 6.50-9.0 . 80% of the activity was retained after 15 min at 50 degrees C . This enzyme produced maltohexaose from starch, amylose and amylopectin by exo-attack, but did not act on alpha- or beta-cyclodextrin, pullulan or maltohexaitol . Also the enzyme acted on beta-limit dextrins of amylopectin and glycogen to form branched oligosaccharides . The unusual reaction of this enzyme on beta-limit dextrin is discussed from the standpoint of the stereochemistry of 1,4-alpha- and 1,6-alpha-glucosidic bonds . This is the anomalous amylase for which it is recognized that 1,6-alpha-glucosidic linkages in the substrates can mimic the effect of 1,4-alpha-bonds, as previously observed in pseudo-priming reactions of E . coli phosphorylase.

J Antibiot (Tokyo), 1975 Dec, 28(12), 943 - 6
DB-2073, a new alkylresorcinol antibiotic . II . The chemical structure of DB-2073; Kitahara T et al.; DB-2073 (I), an antibiotic produced by Pseudomonas sp . B-9004, was obtained as a pure crystals state having a formula of C15H24O2 (ME 236) . The characterization indicated that I was a resorcinol antibiotic and it was elucidated as 2-n-hexyl-5-n-propylresorcinol.

J Bacteriol, 1975 Dec, 124(3), 1177 - 90
Characterization of neutral amino acid transport in a marine pseudomonad; Fein JE et al.; The transport of neutral amino acids in marine pseudomonad B-16 (ATCC 19855) has been investigated . From patterns of competitive inhibition, mutant analysis, and kinetic data, two active transport systems with overlapping substrate specificities were distinguished and characterized . One system (DAG) served glycine, D-alanine, D-serine, and alpha-aminoisobutyric acid (AIB) and, to a lesser extent, L-alanine and possibly other related neutral D- and L-amino acids . The other system (LIV) showed high stereospecificity for neutral amino acids with the L configuration and served primarily to transport L-leucine, L-isoleucine, L-valine, and L-alanine . This system exhibited low affinity for alpha-aminoisobutyric acid . Neither system was able to recognize structural analogues with modified alpha-amino or alpha-carboxyl groups . The kinetic parameters for L-alanine transport by the DAG and LIV systems were determined with appropriate mutants defective in either system . For L-alanine, Kt values of 4.6 X 10(-5) and 1.9 X 10(-4) M and Vmax values of 6.9 and 20.8 nmol/min per mg of cell dry weight were obtained for transport via the DAG and LIV systems respectively . alpha-Aminoisobutyric acid transport heterogeneity was also resolved with the mutants, and Kt values of 2.8 X 10(-5) and 1.4 X 10(-3) M AIB were obtained for transport via the DAG and LIV systems, respectively . Both systems required Na+ for activity (0.3 M Na+ optimal) and in this regard are distinguished from systems of similar substrate specificity reported in nonmarine bacteria.

Ann Surg, 1975 Dec, 182(6), 754 - 61
Infection in war wounds: experience during the 1973 October War in Israel; Simchen E et al.; The development of infections in 420 wounded soldiers, admitted to the Hadassah University Hospital in Jerusalem between October 7, 1973 and November 31, 1973, was studied . An attempt was made to relate the development of infection to the type of injury . The overall infection rate was 22%, but varied with the type of injury . Three "risk factors" were found to be associated with infection regardless of the number of injuries: 1)penetrating abdominal wounds involving the colon; 2) fractures involving the femur; 3) burns involving more than 25% of body surface . In patients with comparable injuries, the presence of infection was found to prolong the duration of hospitalization . Pseudomonas was the most common single pathogen . There were no cases of myonecrosis (gas gangrene) . Of the 8 soliders who died, 5 died with or because of infection.

Br J Ophthalmol, 1975 Dec, 59(12), 725 - 9
Nebcin in the treatment of experimental Pseudomonas keratitis; Belfort R Jr et al.; When the ocular toxicity and the in vivo and in vitro effects of gentamicin, Nebcin, and saline solution were compared in experimentally induced Pseudomonas keratitis in rabbits, both antibiotics showed the same toxicity for the rabbits' conjunctival tissues . But Nebcin showed better in vitro and in vivo results than gentamicin, and the clinical effect was confirmed by culture study: significant numbers of organisms were recovered from the corneas of the gentamicin-treated rabbits but none from the corneas of the Nebcin-treated rabbits.

Biochim Biophys Acta, 1975 Nov 20, 410(1), 12 - 20
Purification and properties of xanthine dehydrogenase from Pseudomonas acidovorans; Sin IL; Xanthine dehydrogenase (EC 1.2.1.37) from Pseudomonas acidovorans has been purified to near homogeneity (approx . 65-fold) . The enzyme has a molecular weight of about 275 000 . Electrophoresis in gels containing sodium dodecyl sulphate showed the presence of two types of subunit with molecular weights of about 81 000 and 63 000 . Thus the intact molecule probably contains two of each type of subunit . Xanthine and hypoxanthine are good substrates, and NAD+ is an effective electron acceptor . With xanthine and NAD+ as substrates the purified enzyme has a specific activity of about 20 mumol NADH formed/min per mg protein . Michaelis constants for xanthine and NAD+ are 0.07 and 0.12 mM, respectively, and for hypoxanthine and NAD+ 0.29 and 0.16 mM, respectively.

Arch Microbiol, 1975 Nov 7, 105(3), 241 - 8
Catabolism of D-fructose and D-ribose by Pseudomonas doudoroffii . II . Properties of 1-phosphofructokinase and 6-phosphofructokinase; Baumann L et al.; 1 . The 1-P-fructokinase (1-PFK) and 6-P-fructokinase (6-PFK) from Pseudmonas doudoroffii were partially purified by a combination of (NH4)2SO4 fractionation and DEAE-Sephadex column chromatography . The pH optima of these enzymes were 9.0 and 8.5, respectively . 2 . When the concentrations of the substrates of the 1-PFK reaction were varied, Michaelis-Menten kinetics were observed . The Kms for D-fructose-1-P (F-1-P) and ATP were 3.03 X 10(-4) M and 3.39 X 10(-4) M, respectively . Variation of MgCl2 at fixed concentrations of F-1-P and ATP resulted in sigmoidal kinetics; about 10 mM MgCl2 was necessary for maximal activity . Activity of 1-PFK was inhibited when the ratio of ATP:Mg++ was higher than 0.5, suggesting that ATP:2Mg++ was the substrate and that free ATP was inhibitory . Although an absolute requirement for K+ or NH4+ could not be demonstrated, these cations stimulated the rate of the reaction . Activity of 1-PFK was not significantly affected by 3 mM AMP, cyclic-AMP, Pi, D-fructose-6-P (F-6-P), ADP, P-enolpyruvate (PEP), pyruvate, citrate, or L-gluamate . 3 . Sigmoidal kinetics were observed for 6-PFK when the concentration of F-6-P was increased and the level of ATP was kept constant . Activity of 6-PFK was increased by ADP, inhibited by PEP, and unaffected by 3 mM AMP, cyclic-AMP, Pi, F-1-P, pyruvate, or citrate.

Arch Microbiol, 1975 Nov 7, 105(3), 225 - 40
Catabolism of D-fructose and D-ribose by Pseudomonas doudoroffii . I . Physiological studies and mutant analysis; Baumann P et al.; Pseudomonas doudoroffii, a strict aerobe of marine origin, was able to utilize fructose and ribose but not glucose, gluconate, or other hexoses, pentoses, or sugar alcohols as sole sources of carbon and energy . Evidence was presented indicating that in this organism fructose was utilized via an inducible P-enolpyruvate: fructose phosphotransferase system (FPTS) which catalyzed the phosphorylation of fructose in the 1 position . The resulting fructose-1-P (F-1-P) was converted to fructose-1,6-P2 (FDP) by means of an inducible 1-P-fructokinase (1-PFK) . The subsequent conversion of FDP to pyruvate involved enzymes of the Embden-Meyerhof pathway (EMP) which, with the exception of glyceraldehyde-3-P dehydrogenase (G3PDH), were constitutive . Two G3PDH activities were detected, one of which was inducible and NAD-dependent while the other was constitutive and NADP-dependent . Cell-free extracts of P . doudoroffii also contained enzymes of the methylglyoxal pathway (MGP) which converted dihydroxyacetone-P to pyruvate . The low specific activities of enzymes of this pathway as compared to the EMP suggested that the major route of FDP catabolism was via the latter pathway . 2 . Ribose catabolism appeared to involve an inducible uptake system and an inducible ribokinase, the resulting ribose-5-P being converted to glyceraldehyde-3-P and fructose-6-P (F-6-P) by means of constitutive activities of the pentose-P pathway . The F-6-P formed as a result of these reactions was converted to FDP by means of a constitutive 6-P-fructokinase (6-PFK) . Since no activity converting fructose or F-1-P to F-6-P could be detected in cell-free extracts of P . doudoroffii, the results suggested that fructose and ribose were catabolized via 1-PFK and 6-PFK, respectively, the two pathways converging at the level of FDP . Further evidence for this suggestion was obtained from a mutant which lacked an NAD-dependent G3PDH, accumulated FDP from both fructose and ribose, and was not able to grow on either of these compounds . 3 . Ribose grown cells had increased amounts of the fructose uptake system and 1-PFK suggesting that a compound (or compounds) common to the catabolism of both fructose and ribose acted as the inducer(s) of these activities . Evidence was presented suggesting that the probable inducer(s) of 1-PFK and FPTS could be FDP, glyceraldehyde-3-P, or dihydroxyacetone-P . 4 . A mutant unable to grow on fructose was characterized and found to lack FPTS while retaining 1-PFK and other enzyme activities of the EMP and MGP, indicating that a functional FPTS was essential for growth on fructose and suggesting that all or most of this sugar was catabolized via F-1-P.

Arch Dis Child, 1975 Nov, 50(11), 886 - 9
Septicaemia from prolonged intravenous infusions; Thong ML et al.; Four cases of septicaemia in children were traced to contaminated intravenous infusions and volume control sets . In each case Pseudomonas cepacia was isolated from multiple blood cultures and from intravenous fluid within the volume control set . The first patient died of septicaemia after a long and complicated postoperative period . The other three patients received appropriate antibiotics after removal of the contaminated intravenous sets and they recovered within 2 weeks.

J Bacteriol, 1975 Nov, 124(2), 679 - 85
Metabolism of naphthalene, 2-methylnaphthalene, salicylate, and benzoate by Pseudomonas PG: regulation of tangential pathways; Williams PA et al.; Naphthalene is metabolized by Pseudomonas PG through 1,2-dihydroxynaphthalene and salicylate to catechol, which is then degraded by the meta pathway . 2-Methylnaphthalene, but not 1-methylnaphthalene, also serves as a growth substrate and is metabolized by the same route, through 4-methylcatechol . The same nonspecific meta pathway enzymes appear to be induced by growth on either naphthalene or 2-methylnaphthalene . The level to which 2-hydroxymuconic semialdehyde hydrolase is induced is low and probably of no metabolic significance . Growth on salicylate or catechol, both intermediates of naphthalene degradation, or benzoate results in induction of the ortho pathway, the alternative route for catechol dissimilation . No induction of 1,2-dihydroxynaphthalene oxygenase was found in salicylate-grown cells . Anaerobic growth on a succinate-nitrate medium in the presence of various inducers indicates that cis, cis-muconate, or one of its metabolites is the inducer of the ortho pathway enzymes . The inducer or inducers of the early enzymes of naphthalene degradation and of the meta pathway enzymes must be an early intermediate of the naphthalene pathway above salicylate.

J Bacteriol, 1975 Nov, 124(2), 1028 - 9
Growth of Pseudomonas C on C1 compounds: a correction; Goldberg I et al.; On reexamination Pseudomonas C was found to be incapable of growth on formaldehyde or formate as a sole carbon source and to contain a hexose phosphate synthase activity when grown on methanol.

J Bacteriol, 1975 Nov, 124(2), 825 - 33
Reduced nicotinamide adenine dinucleotide-activated phosphoenolpyruvate carboxylase in Pseudomonas MA: potential regulation between carbon assimilation and energy production; Newaz SS et al.; Comparison of enzyme activities in crude extracts of methylamine-grown Pseudomonas MA (ATCC 23319) to those in succinate-grown cells indicates the involvement of an acetyl coenzyme A-independent phosphoenolpyruvate carboxylase in one-carbon metabolism . The purified phosphoenolpyruvate carboxylase is activated specifically by reduced nicotinamide adenine dinucleotide (KA = 0.2 mM) . The regulatory properties of this enzyme suggests that phosphoenolpyruvate serves as a focal point for both carbon assimilation and energy metabolism.

Mikrobiologiia, 1975 Nov-Dec, 44(6), 1022 - 4
{Effect of cultivation conditions on cell composition of Pseudomonas methanolica}; Shulgovskaia EM et al.; The effect of cultivation conditions on the ocmposition of polymers was studied in the cells of Pseudomonas methanolica, BKM B-1131 . The economic coefficient was high and the composition of polymers was normal if the growth was limited by methanol deficiency and the flow rate was low (D = 0.1 hr-1) . The composition of the cells did not change but the economic coefficient decreased (0.12--0.21) if the growth was inhibited by pH.

J Gen Microbiol, 1975 Nov, 91(1), 79 - 91
Oxidation of carbon monoxide and methane by Pseudomonas methanica; Ferenci T et al.; The oxidation of carbon monoxide and methane by suspensions and ultrasonic extracts of Pseudomonas methanica was studied . A continuous assay for the oxidation of CO to CO2 was devised, using O2 and CO2 electrodes in combination . Stoicheiometries of CO-dependent CO2 formation, O2 consumption and NADH oxidation, and the partial stoicheiometries of methane-dependent NADH oxidation, suggest the involvement of a mono-oxygenase in these oxidations . Evidence is presented suggesting methane and CO oxidation are catalysed by a single enzyme system, distinct, at least in part, from the NADH oxidase present in extracts . Ethanol was able to provide the reductant necessary for CO oxidation by cell suspensions, though the metabolism of ethanol by P . methanica was found unlikely to result in substrate-level formation of NADH; the means whereby alcohol oxidation could supply reductant for the mono-oxygenase are discussed.

Biochim Biophys Acta, 1975 Oct 22, 403(2), 412 - 24
Studies on phospholipase C from Pseudomonas aureofaciens . I . Purification and some properties of phospholipase C; Sonoki S et al.; Phospholipase C (phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) from Pseudomonas aureofaciens was purified 3600-fold from the culture filtrate with a recovery of 1.6% . Purification was performed with the useof (NH4)2SO4 precipitation, Sephadex G-100 gel filtration and by ion-exchange chromatography on DEAE-Sephadex A-50 and CM-Sephadex C-50 . The purified enzyme appeared to be homogeneous as revealed by polyacrylamide disc gel electrophoresis at pH 9.3 . The molecular weight was estimated to be 35 000 by gel filtration on Sephadex G-75 . Under our experimental conditions, phosphatidylethanolamine was more rapidly hydrolysed than phosphatidylcholine . Lyso forms of these two phosphatides were poor substrates . Phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, cardiolipin and sphingomyelin were not hydrolysed . The enzyme activity with phosphatidylcholine as substrate was slightly stimulated by Ca2+, Mg2+, and Mn2+ . However, these cations inhibited the activity with phosphatidylethanolamine as substrate . An anionic detergent, sodium deoxycholate, slightly enhanced the activity when phosphatidylcholine and phosphatidylethanolamine were used as substrates . A cationic detergent, cetyltrimethylammonium bromide, inhibited enzyme activity . EDTA and o-henanthroline inhibited the activity of the enzyme to a marked degree.

J Clin Microbiol, 1975 Oct, 2(4), 296 - 9
Effects of atmosphere of incubation and of routine subcultures on detection of bacteremia in vacuum blood culture bottles; Harkness JL et al.; Studies comparing isolation rates of bacteria and yeasts from vented and unvented vacuum blood culture bottles containing soybean-casein digest broth showed significantly more frequent and more rapid recovery of Candida and Pseudomonas from the vented bottle and no other statistically significant differences between the two . Subculture of bottles on the day of their collection was shown to accelerate recovery of 48% of positive cultures by day 1 . A second subculture of known positive cultures yielded additional organisms in 1.5% of cultures.

Ann Intern Med, 1975 Oct, 83(4), 512 - 3
Human-to-human transmission of Pseudomonas pseudomallei; McCormick JB et al.; Melioidosis, the clinical manifestation of infection with Pseudomonas pseudomallei, has occurred infrequently in American citizens; almost all reported cases have been in Vietnam veterans, usually associated with respiratory disease . A Vietnam veteran from Mississippi developed chronic prostatitis, with no other clinical manifestations, during service in Vietnam, and P . pseudomallei was isolated from prostatic secretions 2 years after his return to the United States . The patient had had sexual contact with four women including his wife since his return from Vietnam . Vaginal and cervical cultures and serum samples were obtained from the four women, and serum samples and cultures of semen were obtained from the patient . Vaginal swabs and semen cultures were negative for P . pseudomallei . The patient and his wife had hemagglutination titers (greater than 640) diagnostic of P . pseudominallei infection . This occurrence of venereal transmission is the first report of person-to-person spread of P . pseudomallei infection.

Clin Chem, 1975 Oct, 21(11), 1630 - 1
Specificity of oxidation of bile-salt hydroxyl groups by crude extracts of Pseudomonas testosteroni (ATCC 11996) used in determining bile salts; Beher WT et al.; Recent experimental evidence suggests that, presumably as a recent of mutation, crude extracts of Pseudomonas testosteroni (ATCC 11996) no contain amounts of 7alpha- and 12alpha-hydroxysteroid dehydrogenate activity that invalidate the results of bile-salt determinations . To confirm or deny this, we studied the specificity of hydroxysteroid dehydrogenase contained in crude extracts of currently available samples of this bacterium in the oxidation of bile-salt hydroxyl groups . The dehydrogenases in these extracts specifically oxidized 3alpha-hydroxyl groups of cholate, chenodeoxycholate, and deoxycholate anions . No significant amount of 12alpha- or 7alpha-hydroxysteriod dehydrogenase activity was detected . Currently available crude extracts of Pseudomonas testosteroni (ATCC 11996) are therefore suitable for bile-salt quantification.

J Gen Microbiol, 1975 Oct, 90(2), 271 - 85
The respiratory system of Chromobacterium violaceum grown under conditions of high and low cyanide evolution; Niven DF et al.; The particulate fraction of disrupted Chromobacterium violaceum grown under cyanide-evolving conditions was unable to oxidize ascorbate plus N,N,N',N'-tetra-methyl-p-phenylenediamine (TMPD), but oxidized NADH and succinate by a linear respiratory pathway which was very resistant to inhibition by cyanide . When the bacteria were grown under conditions where little cyanide evolution occurred, particulate fractions developed the ability to oxidize ascorbate-TMPD by a pathway highly sensitive to cyanide inhibition; respiratory activity with NADH and succinate proceeded via both the cyanide-sensitive and-resistant pathways . Studies with respiratory inhibitors, and the cytochrome compositions of the fractions derived from cultures grown under both conditions, are presented . A soluble, carbon monoxide-binding cytochrome c was found, and this appears similar to those found recently in Beneckea natiegens, methylotrophic bacteria and the marine pseudomonad B16.

Appl Microbiol, 1975 Oct, 30(4), 650 - 6
Enzymatic transformation of morphine by hydroxysteroid dehydrogenase from Pseudomonas testosteroni; Liras P et al.; Eznyme preparations from Pseudomonas testosteroni containing alpha- and beta- hydroxysteroid dehydrogenases catalyzed the oxidation of morphine and codeine by nicotinamide adenine dinucleotide . Morphine was converted in relatively low yield into 14-hydroxymorphinone probably via morphinone as an intermediate . Codeine was converted to codeinone and 14-hydroxycodeinone . Only the conversions at the 6-position were carred out by the hydroxysteroid dehydrogenase . Hydroxylation at the 14-position did occur spontaneously (or enzymatically with a contaminating enzyme) ater oxidation at the 6-position.

Can J Microbiol, 1975 Oct, 21(10), 1476 - 83
A p-nitrophenyl alpha-galactoside hydrolase from Pseudomonas atlantica . Localization of the enzyme; Day DF et al.; A p-nitrophenyl alpha-galactoside hydrolase is partially released when whole cells of Pseudomonas atlantica are converted to spheroplasts . The p-nitrophenyl alpha-glactoside hydrolase is completely inactivated by treatment of whole cells with diazonaphthalene -- disulfonic acid (NDS), a reagent which does not penetrate the cytoplasmic membrane . Under the conditions used no inactivation of lactic acid dehydrogenase was observed . A specific staining procedure for this enzyme for use in electron microscopy was developed . The results with this technique in conjunction with the results of spheroplasting and NDS localization suggest that p-nitrophenyl alpha-galactoside hydrolase is located in or on the double-track membranes, primarily on the outer double track.

Can J Microbiol, 1975 Oct, 21(10), 1512 - 8
Enzymatic hydrolysis of agar: purification and characterization of neoagarobiose hydrolase and p-nitrophenyl alpha-galactoside hydrolase; Day DF et al.; The mixture of polysaccharides in the gelling component of agar (agarose) is hydrolyzed to D-galactose and 3,6-anhydro-L-galactose by a series of hydrolytic enzymes obtained from Pseudomonas atlantica . The final degradative step in the pathway of agarose decomposition is the hydrolysis of the alpha-linkage in the dissaccharide neoagarobiose yielding D-galactose and 3,6-anhydro-L-galactose . Pseudomonas atlantica when grown on agar produces two specific enzymes, p-nitrophenyl alpha-galactose hydrolase and neoagarobiose hydrolase . The purification and partial characterization of both enzymes are presented.

Biochim Biophys Acta, 1975 Sep 22, 403(1), 245 - 55
Structural effects on the reactivity of substrates and inhibitors in the epoxidation system of Pseudomonas oleovorans; May SW et al.; The epoxidation reaction catalyzed by an enzyme system of Pseudomonas oleovorans exhibits a substrate specificity different from that expected on the basis of chemical reactivity in non-enzymatic epoxidation reactions . Cyclic and internal olefins, aromatic compounds and styrene are not epoxidated . The reactivity of straight chain diolefins is maximal for octadiene and falls off rapidly as the carbon chain is shortened, but decreases only slightly as the chain is lengthened . In contrast, methyl group hydroxylation is less sensitive to decreasing chain length . As a consequence, propylene and 1-butene are hydroxylated but not epoxidated by this enzyme system . With the substrate 1-decene, which is capable of undergoing both epoxidation and hydroxylation, the former reaction predominates . Methyl imidoesters were found to be inhibitors of enzymatic epoxidation, and the potency of a homologous series of imidoester inhibitors was examined . The results parallel the substrate specificity patterns observed, and support the conclusion that the mode of substrate binding severely moderates the inherent chemical reactivity of the activated oxygen in this system . The effect of the bifunctional imidoester, dimethyladipimidate, was also examined and the results compared with those obtained in other investigations.

J Neurol, 1975 Sep 22, 210(3), 219 - 26
{To the differential diagnosis of cranial nerve lesions: the progressive necrotising external otitis (author's transl)}; Draf W et al.; A review of necrotising external otitis, a relatively unknown and dangerous disease, brings out that, initially, it has three characteristics: a granulating necrotising ostitis of the external meatus, extreme pain and a yellowish green secretion . It is always caused by a pseudomonas infection and in almost all cases the patients suffer from diabetes mellitus . If the condition is not recognized in good time and an extensive debridement of the bone involved not performed promptly, ostomyelitis of the base of the skull may follow with involvement of cranial nerves . Severe chronic osteomyelitis of cervical vertebrae occurred in one of our cases . The neurologist must bear this disease in mind in the differential diagnosis when cranial nerves are affected because the nerve disturbances may become evident only after the local condition has subsided or the nerve deficits may be more prominent than and obscure the local ear condition . The most commonly involved nerve is the facial although there may be multiple cranial nerves involved including the third through the twelfth . If the cervical vertebrae become affected there may be nerve root lesions . A torpid meningoencephalitis may also occur . Close cooperation between otologists and neurologists is necessary to recognize and treat these conditions properly.

Med J Aust, 1975 Sep 20, 2(12), 479 - 80
Chronic melioidosis; Smith KV et al.; This report is of a man who suffered from chronic melioidosis contracted in Malaysia . In the course of the disease he had a lobe of a lung resected, developed empyema and, while this was still draining, developed infection in an ankle . Both the empyema thoracis and the ankle infection were due to Pseudomonas pseudomallel . He now appears to be cured, probably by massive doses of tetracycline.

Nucleic Acids Res, 1975 Sep, 2(9), 1459 - 92
Extracellular nucleases of Pseudomonas BAL 31 . I . Characterization of single strand-specific deoxyriboendonuclease and double-strand deoxyriboexonuclease activities; Gray HB Jr et al.; The culture medium of Pseudomonas BAL 31 contains endonuclease activities which are highly specific for single-stranged DNA and for the single-stranded or weakly hydrogen-bonded regions in supercoiled closed circular DNA . Exposure of nicked DNA to the culture medium results in cleavage of the strang opposite the sites of preexisting single-strand scissions . At least some of the linear duplex molecules derived by cleavage of supercoiled closed circular molecules contain short single-stranded ends . Single-strand scissions are not introduced into intact, linear duplex DNA or unsupercoiled covalently closed circular DNA . Under these same reaction conditions, 0X174 phage DNA is extensively degraded and PM2 form I DNA is quantitatively converted to PM2 form III linear duplexes . Prolonged exposure of this linear duplex DNA to the concentrated culture medium reveals the presence of a double-strand exonuclease activity that progressively reduces the average length of the linear duplex . These nuclease activities persist at ionic strengths up to 4 M and are not eliminated in the presence of 5% sodium dodecyl sulfate . Calcium and magnesium ion are both required for optimal activity . Although the absence of magnesium ion reduces the activities, the absence of calcium ion irreversibly eliminates all the activities.

J Biol Chem, 1975 Sep 10, 250(17), 6875 - 9
The sterochemistry at carbon 3 of pyruvate lyase condensation products . 2-Keto-3-deoxygluconate 6-phosphate and 2-keto-3-deoxygalactonate-6-phosphate aldolase of Pseudomonas saccharophila; Meloche HP et al.; In Pseudomonas saccharophila 2-keto-3-deoxygalactonate-6-P aldolase (EC 4.1.2.21) is induced by growth on galatose while 2-keto-3-deoxygluconate-6-P aldolase (EC 4.1.2.14) is constitutive . These enzymes catalyze identical reactions except for the configuration fixed at C-4 during the condensation reaction . It was found with each enzyme that in a condensation between {3-3H3}pyruvate and D-glyceraldehyde-3-P, the respective condensation products were formed 8 to 10 times faster than tritium was released to water . Since pyruvate deprotonation is obligatory for condensation, the above result requires a hydrogen isotope effect in enolpyruvate formation, which must be then at least partially rate limiting for C--C synthesis . Further, condensation between D-glyceraldehyde-3-P and (3R)-{3-3H, 2H,H}pyruvate or (3S)-{3-3H, 2H,H}pyruvate, as catalyzed by each enzyme, enriched for (3R)- and (3S)-3-3H, 2H-labeled condensation product, respectively . Thus, each enzyme catalyzes C--C and C--H synthesis with retention of configuration at C-3 . This shows that the active sites of both enzymes are asymmetric since solutes can only approach a single face of the bound pyruvyl enolate . In addition, the respective aldehyde specific portions of the two active sites must have opposite chiralities, with respect to each other, for correctly orienting the carbonyl faces of the incoming D-glyceraldehyde-3-P, to generate the correct configuration at C-4 of the respective condensation products.

J Biol Chem, 1975 Sep 10, 250(17), 6672 - 8
Phenylalanine hydroxylase from Pseudomonas sp . (ATCC 11299a) . Purification, molecular weight, and influence of tyrosine metabolites on activation and hydroxylation; Letendre CH et al.; Phenylalanine hydroxylase from Pseudomonas sp . (ATCC 11299a) has been purified 25- to 30-fold by a procedure which has been modified from that previously described for this organism (Guroff, G., and Ito, T . (1965) J . Biol . Chem . 240, 1175-1184; Guroff, G., and Rhoads, C . A . (1967) J . Biol . Chem . 242, 3641-3645) . Further purification yielded a preparation which was judged to be about 80% pure by sodium dodecyl sulfate-containing and standard analytical polyacrylamide gels, but the activity in this preparation has proved to be very labile . The enzyme appears to be a single protein chain of between 25,000 to 27,000 molecular weight . Phenylalanine, tyrosine, and tryptophan inhibit the activation of the enzyme by iron in a competitive fashion . The tyrosine metabolites, p-hydroxyphenylpyruvic and homogentisic acids exhibit a biphasic effect on activation, stimulating at low iron, and inhibiting at higher iron concentrations . The hydroxylation itself is inhibited by tyrosine and related compounds such as L-3,4-dihydroxyphenylalanine and dopamine . p-Hydroxyphenylpyruvic acid is a competitive inhibitor with respect to both substrate and cofactor . The data indicate a variety of means by which the bacterium can regulate phenylalanine hydroxylation.

Mikrobiologiia, 1975 Sep-Oct, 44(5), 779 - 83
{Hydrogenase activity of the methylotroph, Pseudomonas methylica}; Gogotov IN et al.; The cells of Pseudomonas methylica, strain 2, cultivated in a medium containing methanol, displayed the activity of hydrogenase in the exchange reaction (D2--H2O) and in the absorption of H2 in the presence of methylviologen, azocarmine, methylene blue, and ferricyanide . The rate of H2 utilization was highest in the presence of methylviologen . Cell extracts absorb H2 in the presence of methylviologen, NAD, and NADP, but much faster in the presence of flavin mononucleotide . The bulk of the hydrogenase remains, during centrifugation of the initial cell extract (3,000 g), in the soluble fraction (144,000 g) . The absorption of oxygen by the cell suspensions and the incorporation of 14C of formiate into the cells are stimulated by H2 . The cells, however, cannot grow in the autotrophic conditions at the account of molecular hydrogen and CO2.

J Pediatr, 1975 Sep, 87(3), 474 - 9
Clinical pharmacology of ticarcillin in the newborn infant: relation to age, gestational age, and weight; Nelson JD et al.; Ticarcillin, a new penicillin derivative with greater in vitro activity against Pseudomonas than carbenicillin, was given as a single dose of 50 mg/kg intramuscularly on 61 occasions to 54 neonates and young infants . The serum concentrations and rate of elimination varied considerably depending upon gestational age, chronological age, and body weight . Peak serum concentrations depended principally upon the volume of distribution which approximated the extracellular fluid volume of the neonate . The plasma clearance and serum half-life of the drug were determined . From these pharmacokinetic data appropriate groupings according to age and body weight were made and dosages and intervals of administration predicted for clinical trials of efficacy with repeated doses.

J Clin Pathol, 1975 Sep, 28(9), 741 - 3
Meningitis caused by Pseudomonas maltophilia; Patrick S et al.; A case of meningitis caused by Pseudomonas maltophilia is described, which was unusual in that it appeared to lack the predisposing factors commonly associated with this organism . Attention is drawn to the difficulties which may be encountered in the identification of Ps . maltophilia.

Am J Clin Pathol, 1975 Sep, 64(3), 382 - 4
Respiratory colonization with Pseudomonas putrefaciens after near-drowning in salt water; Rosenthal SL et al.; Pseudomonas putrefaciens, a marine organism infrequently found in human culture material, was repeatedly isolated from the sputum of a patient with pneumonia during a three-week period following a salt-water drowning accident . Similar organisms were found in the water at the site of the accident in Boston, and at ocean bathing beaches on nearby Martha's Vineyard.

J Virol, 1975 Sep, 16(3), 685 - 95
Proteins of bacteriophage phi6; Sinclair JF et al.; We investigated the protein composition of the lipid-containing bacteriophage phi 6 . We also studied the synthesis of phage-specific proteins in the host bacterium Pseudomonas phaseolicola HB10Y . The virion was found to contain 10 proteins of the following molecular weights: P1, 93,000; P2, 88,000; P3, 84,000; P4, 36,800; P5, 24,000; P6, 21,000; P7, 19,900; P8, 10,500; P9, 8,700; and P10, less than 6,000 . Proteins P3, P9, and P10 were completely extracted from the virion with 1% Triton X-100 . Protein P6 was partially extracted . Proteins P8 and P9 were purified by column chromatography . The amino acid composition of P9 was determined and was found to lack methionine . Labeling of viral proteins with {35S}methionine in infected cells indicated that proteins P5, P9, P10, and P11 lacked methionine . Treatment of host cells with UV light before infection allowed the synthesis of P1, P2, P4, and P7; however, the extent of viral protein synthesis fell off exponentially with increasing delay time between irradiation and infection . Treatment of host cells with rifampin during infection allowed preferential synthesis of viral proteins, but the extent of synthesis also fell off exponentially with increasing delay time between the addition of rifampin and the addition of radioactive amino acids . All of the virion proteins were seen in gels prepared from rifampin-treated infected cells . In addition, two proteins, P11 and P12, were observed; their molecular weights were 25,200 and 20,100, respectively . Proteins P1, P2, P4, and P7 were synthesized early, whereas the rest began to increase at 45 min post-infection.

J Bacteriol, 1975 Sep, 123(3), 954 - 61
Regulation of extracellular protease secretion in Pseudomonas maltophilia; Boethling RS; Cells grown in minimal medium and harvested in late exponential phase secreted extracellular protease linearly when suspended at high density in fresh medium . If the cells were suspended with 0.2% (wt/vol) yeast extract and no additional carbon source, the rate of exoenzyme production was increased several-fold . When pyruvate, L-malate, succinate, or alpha-ketoglutarate was added, repression of exoenzyme secretion was observed . The most effective repressor was alpha-ketoglutarate . These compounds were also preferred substrates for growth of Pseudomonas maltophilia . The data suggest that exoenzyme secretion is controlled by a mechanism similar to catabolite repression . In support of this was the observation that alpha-ketoglutarate repressed exoenzyme secretion preferentially with respect to total protein synthesis . The addition of inhibitors that affect protein synthesis indicated that exoenzyme secretion is several times more sensitive than is total protein synthesis . The addition of chloramphenicol and rifamycin-SV to actively secreting cell suspensions suggested that de novo protein synthesis is required, but that exoenzyme secretion may be supported for at least 30 min in the absence of messenger synthesis . Rifamycin-insensitive protease secretion could be reversed by either alpha-ketoglutarate or chloramphenicol, suggesting that alpha-ketoglutarate is coupled to a post-transcriptional control mechanism.

Proc Soc Exp Biol Med, 1975 Sep, 149(4), 845 - 8
Endotoxin-induced release of colony-stimulating activity in man; Golde DW et al.; Six healthy adult volunteers were injected intravenously with Pseudomonas endotoxin (Piromen) to determine effects on serum colony-stimulating activity (CSA) . Serum was assayed with a double-layer agar cloning technique using normal human bone marrow as the target cell preparation . A marked increase in serum CSA was noted after endotoxin administration lasting up to 180 min . The colonies formed in response to postendotoxin serum were smaller than those seen with leukocyte feeder layers but were otherwise morphologically normal . These studies suggest that granulopoietic humoral factors may be modulated in man by endotoxin administration.

Mikrobiologiia, 1975 Sep-Oct, 44(5), 839 - 43
{Effect of H+ and OH- ions on the physiological and biochemical properties of a Pseudomonas methanolica culture}; Ivanova II et al.; The growth of Pseudomonas methanolica was inhibited by unfavourable values of pH in the conditions of chemostat; the rate of the substrate assimilation was higher, and anabolic and catabolic reactions were decoupled . Hydrogen ions inhibited the activity of enzymes of the Krebs cycle, hydroxyl ions inhibited the activity of methanol dehydrogenase . Changes in pH are presumed to involve the energy apparatus of the cell membrane.

Mol Cell Biochem, 1975 Aug 30, 8(2), 113 - 21
Hydroxyproline-2-epimerase of Pseudomonas: active-site peptides; Zervos C et al.; Hydroxyproline-2-epimerase was treated with 14C-iodoacetate under conditions that produced almost complete inactivation of the enzyme and concomitant incorporation of almost one molar equivalent of iodoacetate . Both processes were prevented by saturating concentrations of substrate . From reaction mixtures in which both incorporation and inactivation were 85 to 90% complete, two radioactive tryptic peptides were isolated by paper chromatography-electrophoresis . The incorporated radioactivity was divided between the peptides in an approximately 2:1 ratio . Analysis of the isolated peptides suggested that they both contained 9 amino acids and had similar composition; one appeared to be a lysine, the second an arginine peptide . Attempts to sequence each peptide failed, apparently because of the conversion of the S-carboxymethylcysteine to S-carboxymethylcysteine sulfone, indicating that the cysteine residue was N-terminal in each peptide.

J Biol Chem, 1975 Aug 25, 250(16), 6208 - 13
pH dependence of the cooperative interactions and conformation of tryptophan oxygenase; Colman PD et al.; Allosteric interactions in the cupro-heme enzyme tryptophan oxygenase (EC 1.13.11.11) of Pseudomonas acidovorans are shown to be pH-dependent . Increasing the assay pH from 6.0 to 8.0 progressively desensitizes the enzyme from both homotropic and heterotropic ligand interactions . This pH-dependent reversible transition has a pK of 6.2 . Hill coefficients for the substrate L-tryptophan of 2.0 and 1.4 were measured at pH 6.0 and pH 7.0, respectively . In attempting to identify the enzymatic residue (or residues) responsible for these pH-dependent effects, the enzyme was observed to be irreversibly inactivated by photoinduced oxidation in the presence of the sensitizer, methylene blue . The photoinactivated enzyme showed a loss of one-half its Soret (405 nm) absorption which accompanied the loss of one-half its heme and histidine contents . This first order photoinduced inactivation was pH-dependent and corresponded to a requirement for a protonated species with a pK of 6.2 . These results suggest that histidine residues may be involved in the catalytic function and in mediating cooperative interactions of tryptophan oxygenase . Absolute and difference sedimentation velocity analyses indicate that the molecule undergoes a conformational transition when the pH is decreased from pH 8.0 to pH 6.0 . This conformational alteration, measured as a 3.9% increase in S20, w can be regarded as an equivalent decrease in the frictional coefficient . If, a more or less spherical shape to the molecule is assumed, then, the 3.9% decrease in the frictional coefficient between pH 8.0 and 6.0 corresponds to a 12% decrease in apparent hydrodynamic volume of the enzyme . Thus, protonation of an enzymatic moiety, possibly histidine, determines both the conformational and functional interactions between enzymatic sites.

Biochemistry, 1975 Aug 12, 14(16), 3682 - 7
Bromopyruvate inactivation of 2-keto-3-deoxy-6-phosphogalactonate aldolase of Pseudomonas saccharophila . Kinetics and stereochemistry; Meloche HP et al.; The enzyme 2-keto-3-deoxy-6-phosphogalactonate aldolase of Pseudomonas saccharophila is inactivated by the substrate analog beta-bromopyruvate, which satisfies several criteria of being an active site directed reagent . The inactivation exhibits saturation kinetics, and both bromopyruvate and pyruvate (substrate) compete for free enzyme . Upon prolonged incubation, inactivation is virtually complete . The Kinact for bromopyruvate is 12 mM and the minimum inactivation half-time is 16 min with a k of 0.0433 min minus 1 . Bromopyruvate is also a substrate for the enzyme in that 3(R,S)-{3-3H2}bromopyruvate is asymmetrically detritiated by the enzyme yielding 3(S)-{3-3H,H}bromopyruvate concomitant with inactivation . At various concentrations of bromopyruvate which affect the inactivation rate, the ratio of nanomoles of bromopyruvate turned over/unit of enzyme inactivated remains constant averaging 12:1, consistent with both inactivation and catalysis occurring at a single protein site, the catalytic site . The above value does not take into account a possible hydrogen isotope effect and is not thus an absolute value . The stereochemistry of bromopyruvate turnover catalyzed by this enzyme is the same as that for 2-keto-3-deoxy-6-phosphogluconate aldolase of P . putida . This fact provides the first evidence that the pyruvate-specific portions of the two active sites may have evolved from a common precursor.

Aust Vet J, 1975 Aug, 51(8), 395 - 8
Bovine melioidosis in South-Eastern Queensland; Ketterer PJ et al.; Acute fatal infection with Pseudomonas pseudomallei caused pneumonia, placentitis and endometritis in a pregnant cow . Pneumonia was also present in the foetal lung . Mononuclear cell response with extensive karyorrhexis occurred in maternal and foetal lung and the same cellular pathology, but with diffuse distribution, was responsible for plaques on the endometrium . A more chronic disease with encapsulated caseous lesions in the lung, together with arthritis, occurred in a bull on the same property . Nutritional and physical stress caused by a flood were thought to be predisposing factors . An unusually wet summer with prolonged flooding apparently provided suitable environmental conditions for saprophytic growth of Ps . pseudomallei in an area far south of the region in which infection of animals with this organism commonly occur.

Arzneimittelforschung, 1975 Aug, 25(8), 1318 - 9
{Studies on the influence of pentosanepolysulfate and a combination of pentosanepolysulfate, metamizol and paracetamol on inflammatory liability (author's transl)}; Feyen H et al.; The influence of pentosanepolysulfate (SP 54) and a combination of pentosanepolysulfate, metamizol and paracetamol (Probaphen) on the course of an experimentally induced erythema caused by intracutaneous injection of a lipopolysaccharide from Pseudomonas has been studied in man . The intramuscular injection of Probaphen as well as SP 54, separately given, showed a significant antiinflammatory effect . A similar effect was also achieved after administration of Probaphen suppositories . Probaphen shortened the inflammatory reaction more effectively than did SP 54 alone . The oral application of Probaphen, however, did not influence the course of inflammation.

Eur J Biochem, 1975 Jul 15, 55(3), 601 - 9
Inhibition by carbon monoxide of the secondary-amine mono-oxygenase of Pseudomonas aminovorans and the photochemical action spectrum for its reversal; Brook DF et al.; The secondary-amine mono-oxygenase (EC 1.14.99.--) of Pseudomonas aminovorans is potently inhibited by carbon monoxide . The degree of inhibition of the purified enzyme was determined by the CO:O2 ratio rather than by the absolute concentration of carbon monoxide . The partition constant (the CO:O2 ratio causing 50% inhibition of activity) was 9.2 X 10(-4) . The inhibition could be reversed by light, and the extent of reversal was proportional to the light intensity . With monochromatic light of wavelength 417 nm, the light sensitivity, L, was determined to be 2.5 X 10(8) cm2 min/mol quantum . The photochemical action spectrum for the light reversal of inhibition showed a single maximum of effectiveness at about 420 nm . The difference spectrum of the enzyme (reduced by NADH) on bubbling with CO (compared with an NADH-reduced reference sample) showed a peak at 426 nm . The preparations showed none of the spectral properties to cytochrome P-450 mono-oxygenase preparations, and was much more sensitive to carbon monoxide . The enzyme behaves as a typical o-type cytochrome (i.e . a carbon-monoxide-reactive b-type cytochrome), and its sensitivity to carbon monoxide as well as in its spectral properties, shows close resemblances to haemoglobin.

J Biol Chem, 1975 Jul 10, 250(13), 5041 - 8
The oxygenated complexes of the two catalytically active oxidation-reduction states of L-tryptophan-2,3-dioxygenase; Brady FO et al.; The oxygenated complexes of the two catalytically active forms of pseudomonad and rat liver L-tryptophan-2,3-dioxygenase (EC 1.13.11.11) have been studied . As was previously reported (ISHIMURA, Y., NORZAKI, M., HAYAISHI, O., TAMURA, M., AND YAMAZAK-I I . (1970) J . Biol . Chem . 245, 3593-3602), we observe that the fully reduced form of pseudomonad tryptophan oxygenase during steady state catalysis exists predominantly as the L-tryptophan ferroheme-O2 enzyme complex (lambdamax = 415 nm, 540 nm, 570 nm) . However, during steady state catalysis by a half-reduced form of both the pseudomonad and hepatic enzymes, the predominant species present manifest absorption spectra indicative of ternary complexes in which all the heme exists as ferriheme (Soret, 407 nm), there being no trace of a ferroheme-O2 complex . Carbon monoxide is a competitive inhibitor with respect to molecular oxygen of catalysis by either the half-reduced or fully reduced forms of pseudomonad tryptophan oxygenase . During steady state catalysis in the presence of CO, the fully reduced form of the enzyme exists as a mixture of the oxyferroheme (Soret = 415 nm) and carboxyferroheme (Soret = 421 nm) enzyme complexes . However, if the same experiment is repeated with the half-reduced form of the pseudomonad enzyme, all of the enzyme is in the ferriheme state, even though CO is inhibiting this form of the enzyme to the same degree as it does the fully reduced form . We conclude that for the half-reduced form of pseudomonad tryptophan oxygenase the substrate, O2, and the inhibitor, CO, are not binding to the heme moieties, but are bound elsewhere, presumably to the Cu(I) moieties . Examination of the kinetic mechanisms of the half-reduced and fully reduced forms of pseudomonad tryptophan oxygenase using the inhibitors carbon monoxide and 5-fluorotryptophan confirmed that the fully reduced enzyme binds L-tryptophan before O2 (FORMAN, H., AND FEIGELSON, P . (1971) Biochemistry 10, 760-763) and that for the half-reduced enzyme O2 binds first . In the presence of 5-fluorotryptophan a relatively stable oxyferroheme enzyme complex was generated with the fully reduced form of pseudomonad tryptophan oxygenase . Thus, saturation of the catalytic site alone either with the substrate, L-tryptophan, or the competitive inhibitor, 5-fluorotryptophan, enhances binding of O2 to the ferroheme moieties of the enzyme . The resistance of this complex to photolysis indicates that the bound molecular oxygen is predominantly present as superoxide, O2-minus.

J Biol Chem, 1975 Jul 10, 250(13), 5233 - 42
Structural, kinetic, and renaturation properties of an induced hydroxypyruvate reductase from Pseudomonas acidovorans; Utting JM et al.; A hydroxypyruvate reductase has been induced in Pseudomonas acidovorans by growth on glyoxylate . The enzyme has been purified to homogeneity as assessed by the criteria of analytical ultracentrifugation and analytical disc gel electrophoresis . It has a molecular weight of approximately 85,000 and is composed of two identical subunits . The subunits are not interconnected by disulfide bonds although the enzyme has 4 mol of half-cystine per mol of enzyme . The enzyme catalyzes the reversible conversion of hydroxypyruvate to D(minus)-glycerate in the presence of NADH . Glyoxylate cannot replace hydroxypyruvate as a substrate and is a competitive inhibitor of hydroxypyruvate reduction . The activity of the enzyme toward hydroxypyruvate is anion-modulated; the activity of the enzyme toward D(minus)-glycerate is unaffected by anions but is increased by tris-(hydroxymethyl)aminomethane . The subunits of the induced hydroxypyruvate reductase can be renatured . After the enzyme is dissociated in solutions of 6.0 M guanidine hydrochloride containing 0.1 M 2-mercaptoethanol, optimum renaturation occurs when subunits are diluted into a renaturation solvent consisting of 0.04 M Trischloride, pH 7.4, containing 25% glycerol, 25 mM 2-mercaptoethanol, and 0.14 MM NADH . NAD is an inhibitor of renaturation and therefore cannot substitute for NADH . The optimal temperature of dilution and subsequent incubation is 15 degrees, and increases in protein concentration up to 1.2 mg/ml, the highest concentration tested, improve both the rate of renaturation and the yield of active material . The half-time of renaturation at a protein concentration of 1.2 mg/ml was 1 min . The kinetics of renaturation is second order, i.e., is compatible with a bimolecular reaction preducted by the association of two similar subunits . The physical and kinetic parameters of the renatured protein are the same as those of the native enzyme.

J Biol Chem, 1975 Jul 10, 250(13), 4848 - 55
Extradiol cleavage of 3-substituted catechols by an intradiol dioxygenase, pyrocatechase, from a Pseudomonad; Fujiwara M et al.; Pyrocatechase (catechol 1,2-oxidoreductase (decyclizing), EC 1.13.11.1), a ferric ion-containing dioxygenase from Pseudomonas arvilla C-1, catalyzes the intradiol cleavage of catechol with insertion of 2 atoms of molecular oxygen to form cis,cis-muconic acid . The enzyme also catalyzed the oxidation of various catechol derivatives, including 4-methylcatechol, 4-chlorocatechol, 4-formylcatechol (protocatechualdehyde), 4,5-dichlorocatechol, 3,5-dichlorocatechol, 3-methylcatechol, 3-methoxycatechol, and 3-hydroxycatechol (pyrogallol) . All of these substrates gave products having an absorption maximum at around 260 nm, which is characteristic of cis,cis-muconic acid derivatives . However, when 3-methylcatechol was used as substrate, the product formed showed two absorption maxima at 390 and 260 nm . These two absorption maxima were found to be attributable to two different products, 2-hydroxy-6-oxo-2,4-heptadienoic acid and 5-carboxy-2-methyl-2,4-pentadienoic acid (2-methylmuconic acid) . The former was produced by the extradiol cleavage between the carbon atom carrying the hydroxyl group and the carbon atom carrying the hydroxyl group and the carbon atom carrying the methyl group; the latter by an intradiol cleavage between two hydroxyl groups . Since these products were unstable, they were converted to and identified as 6-methylpyridine-2-carboxylic acid and 2-methylmuconic acid dimethylester, respectively . Similarly, 3-methoxycatechol gave two products, namely, 2-hydroxy-5-methoxycarbonyl-2,4-pentadienoic acid and 5-carboxy-2-methoxy-2,4-pentadienoic acid (2-methoxymuconic acid) . With 3-methylcatechol as substrate, the ratio of intradiol and extradiol cleavage activities of Pseudomonas pyrocatechase during purification was almost constant and was about 17 . The final preparation of the enzyme was homogeneous when examined by disc gel electrophoresis and catalyzed both reactions simultaneously with the same ratio as during purification . All attempts to resolve the enzyme into two components with separate activities, including inactivation of the enzyme with urea or heat, treatment with sulfhydryl-blocking reagents or chelating agents, and inhibition of the enzyme with various inhibitors, proved unsuccessful . These results strongly suggest that Pseudomonas pyrocatechase is a single enzyme, which catalyzes simultaneously both intradiol and extradiol cleavages of some 3-substituted catechols.

J Bone Joint Surg Am, 1975 Jul, 57(5), 631 - 5
Septic arthritis due to pseudomonas in heroin addicts; Gifford DB et al.; Ten cases of septic arthritis due to Pseudomonas occurred in users of heroin . Nine cases were monarticular, two each occurring in the sternoclavicular and the sacro-iliac joints . Review of the literature suggests that the incidence of Pseudomonas septic arthritis is increasing, especially among heroin addicts . The reported cases in adults suggest a predilection for the sternoclavicular joints (eight of forty-one) which is even more pronounced among addicts (seven of twenty-four) . Pseudomonas infection associated with drug abuse should be suspected in cases of isolated sternoclavicular inflammation . The drugs used for treatment of septic arthritis due to Pseudomonas should include gentamicin and carbenicillin, in conjunction with adequate drainage by aspiration or surgery.

Can J Microbiol, 1975 Jul, 21(7), 1004 - 8
The bacterial degradation of fluoranthene and benzo{alpyrene; Barnsley EA; A gas chromatographic method to determine fluoranthene and benzo{a}pyrene at concentrations close to the limit of their solubility in water was used to identify several species of pseudomonads able to degrade fluoranthene and benzol{a}pyrene . Degradation occurs most rapidly in cultures in the stationary phase, is heat sensitive, requires oxygen, and is enhanced in the presence of cyanide.

Mikrobiologiia, 1975 Jul-Aug, 44(4), 651 - 6
{Physiological and biochemical properties of a Pseudomonas methanolica culture under chemostat cultivation}; Ivanova II et al.; The effect of different concentrations of methanol on Pseudomonas methanolica BKM B-1131 was studied . The limitation of growth by methanol in chemostat was found at a concentration of the substrate below 0.1--0.23 per cent; higher concentrations resulted in the appearance of residual methanol in the medium and in a decrease of the economic coefficient . A declining chemostat curve has been obtained when the growth was limited by a deficiency of methanol . If the growth rate increased, the content of RNA remained the same, the content of protein decreased from 50 to 39 per cent, and the content of POBA increased . Changes in the specific activity of some key enzymes of the Krebs and Embden-Meyerhof cycles and of methanol dehydrogenase suggest an increase in the activity of the Krebs cycle enzymes with the growth rate because at this time the substances between acetate and pyruvate are accumulated which are favourable substrates for the synthesis of POBA.

Mikrobiologiia, 1975 Jul-Aug, 44(4), 761 - 2
{Acetylcholinesterase activity of bacteria of the genus Pseudomonas}; Velikanov NL et al.; The activity of acetylcholine esterase was determined in 50 strains of the Pseudomonas genus: Ps . aurantiaca (6 strains), Ps . aeruginosa (8 strains), Ps . fluorescens (31 strains), Ps . putida (1 strain), Ps . ovalis (1 strain), Ps . aureofaciens (1 strain), Ps . geniculata (1 strain), and Ps . nonliquefaciens (1 strain) . The active strains of Ps . aeruginosa and Ps . aurantiaca are described.

Am J Med, 1975 Jul, 59(1), 29 - 36
Subacute and acute endocarditis due to Pseudomonas cepacia in heroin addicts; Noriega ER et al.; Five heroin addicts were treated for endocarditis caused by Pseudomonas cepacia . Two of these infections occurred in patients with no known heart disease whereas the others occurred at sites of previous endocarditis or valve prostheses . Infection was indolent in four patients but was associated with shock and skin lesions suggestive of ecthyma gangrenosum in the fifth . After failure of chloramphenicol and kanamycin, all patients were treated with a combination of sulfamethoxazole, trimethoprim and polymyxin plus heart valve resection or replacement.

Thoraxchir Vask Chir, 1975 Jun, 23(3), 187 - 91
{Surgical treatment of acute dissection of the ascending aorta (author's transl)}; Seybold-Epting W et al.; Since 1959, 51 patients underwent open heart surgery for correction of acute dissection of the ascending aorta . Upon admission 33 patients were severely hypotensive or in porgressive heart failure . Acute aortic insufficiency was found in 24 patients, and hemiplegia or hemiparesis in 4 . In 45 patients the ascending aorta was reconstructed with a woven graft . After excision of the dissected part of the aorta primary anastomosis or patch aortoplasty was performed in 6 patients . The aortic valve remained intact in 26 cases, and resuspension of the commissures restored competence of the aortic valve in another 9 patients . Sixteen patients required aortic valve replacement because of disrupture of the commissures . Dissection extended into the coronary ostia in 5 cases.Reconstruction of the coronary system was accomplished by reimplantation of the ostia, interposition of a vein graft or aorto-coronary bypass . Nine patients died within the early postoperative course from uncontrollable hemorrhage (4), further dissection (3) and myocardial infarction (2) . Within the first year after surgery another 5 patients died from acute aortic dissection (2), pseudomonas-infection causing rupture of the proximal graft anastomosis (1), and myocardial infarction (2) . The contraindications of antihypertensive treatment of actue dissection of the ascending aorta are discussed . We recommend prompt surgical intervention in acute dissecting aneurysm of the ascending aorta.

J Bacteriol, 1975 Jun, 122(3), 905 - 10
Enzymes involved in the assimilation of one-carbon units by Pseudomonas MS; Wagner C et al.; Pseudomonas MS can grow on methylamine and a number of other compounds containing C1 units as a sole source of carbon and energy . Assimilation of carbon into cell material occurs via the "serine pathway" since enzymes of this pathway are induced after growth on methylamine, but not malate or acetate . A mutant has been isolated which is unable to grow on methylamine or any other related substrate providing C1 units . This mutant is also unable to grow on acetate . Measurment of enzyme activities in cell-free extracts of wild-type cells showed that growth on methylamine caused induction of isocitrate lyase, a key enzyme in the glyoxylate cycle . The mutant organism lacks malate lyase, a key enzyme of the serine pathway, and isocitrate lyase as well . These results suggest that utilization of C1 units by Pseudomonas MS results in the net accumulation of acetate which is then assimilated into cell material via the glyoxylate cycle.

Biochim Biophys Acta, 1975 May 30, 393(1), 48 - 54
The subunit structure of Pseudomonas cytochrome oxidase; Kuronen T et al.; Pseudomonas cytochrome oxidase (EC 1.9.3.2) is composed of two subunits . Each subunit has a molecular weight of approx . 63000 and, according to the iron determination, contains two hemes . Cytochrome oxidase was subjected to various dissociation procedures to determine the stability of the dimeric structure . Progressive succinylation of 14 to 68% of the lysine residues of the enzyme increases the amount of the protein appearing in the subunit form (S20,W approximately 4 S) from 18 to 92% . At a high degree of succinylation a component with a sedimentation coefficient of approx . 2 S appears . The subunits with sedimentation coefficients of approx . 4 S and 2 S are also formed when the pH is below 4 or above 11 . The same molecular weight (63000) was found for these two components in sodium dodecylsulphate electrophoresis . No dissociation of cytochrome oxidase was observed in salt solutions like 3 M NaC1 and 1 M Na2SO4, or in 6 M urea . The slight decrease in the sedimentation coefficients in NaC1 solutions is partly explained by preferential hydratation of the protein.

J Biol Chem, 1975 May 25, 250(10), 3746 - 51
Purification and characterization of N-methylalanine dehydrogenase; Lin MC et al.; Cell free extracts of Pseudomonas MS previously have been shown to carry out the synthesis of a novel amino acid, N-methylalanine (Kung, H.F., and Wagner, C . (1970) Biochim . Biophys . Acta 201, 513-516) . An enzyme has been isolated from this organism which is responsible for the synthesis of N-methylalanine . The stoichiometry of the reaction catalyzed by this enzyme leads to the following formulation: Methylamine + pyruvate + NADPH + H-+ yields N-methylalanine + NADP-+ + H2O . This enzyme has been physically separated from alanine dehydrogenase, which is also present in these extracts . This new enzyme has been named N-methylalanine dehydrogenase . It has been purified to near homogeneity as judged by disc gel electrophoresis . Gel filtration chromatography showed that N-methylalanine dehydrogenase has an apparent molecular weight of 77,000, while electrophoresis in sodium dodecyl sulfate gave rise to a single band with a molecular weight of approximately 36,500 . The enzyme is optimally active in the pH range between 8.2 and 8.6 . The apparent K-m values for pyruvate, NADPH, and methylamine, respectively, are 1-5 times 10 minus 2 M, 3-5 times 10 minus 5 M, and 7.5 times 10 minus 2 M.

Biochim Biophys Acta, 1975 May 23, 391(1), 96 - 108
A Pseudomonas intracellular amylase with high activity on maltodextrins and cyclodextrins; Kato K et al.; An intracellular amylase from Pseudomonas MSl was purified 95-fold in a 14% yield by fractionation with (NH4) 2SO4, followed by chromatographies on CM-cellulose, hydroxylapatite and Sephadex G-200 . On polyacrylamide gel electrophoresis at pH 8.3 it gave a single band and its molecular weight was determined to be 96 000 from its mobility on sodium dodecylsulfate gel electrophoresis . Its activity was maximal at pH 5.5 and 50 degrees C . This enzyme hydrolyzed maltotriose and maltotetraose faster than amylose, but did not hydrolyze maltose . It also rapidly hydrolyzed a series of cyclomaltodextrins . Amylopectin and glycogen, which have branched structures, were attacked much slower than amylose . The enzyme cleaved the substrates in an endo-wise fashion and produced almost equimolar amounts of glucose and maltose as the final products from various substrates . The enzyme mechanism is discussed on the basis of quantitative autoradiographic data . The K-m values for maltotriose, maltotetraose, cyclomaltohexaose, corn amylose and waxycorn amylopectin were 1.0, 1.4, 3.5, 3.3 and 13 - 10-minus 3 g/ml, respectively . The products of the enzyme had an alpha-configuration . Thus, this enzyme is a specific alpha-amylase.

Eur J Biochem, 1975 May 6, 53(2), 357 - 69
Characterization of an inducible amidase from Pseudomonas acidovorans AE1; Alt J et al.; The main molecular and catalytic properties of an acetanilide-hydrolyzing enzyme from Pseudomonas acidovorans AE 1, purified to a homogeneous state, were investigated . The molecular weight was 57 500 as determined by gel filtration and 55 300 as computed from the amino acid composition . By polyacrylamide gel electrophoresis in dodecylsulfate a polypeptide chain weight of 56 700 was obtained . Based on the reaction of the highly purufied enzyme with diethyl-4-nitrophenyl phosphate an equivalent weight of approximately 59 100 was found . From these results it was concluded that the enzyme consists of a single polypeptide chain and contains one active site per molecule . The enzyme hydrolyzed esters as well as certain aromatic amides . It also catalysed the transfer of acetyl groups to phenetidine yielding phenacetin . The activities towards aliphatic esters were much smaller . The enzyme was stable at pH values ranging from 7 to 9 and its pH-optimum was about 10 . It was strongly inhibited by organophosphorous compounds, like diethyl-4-nitrophenyl phosphate or diisopropylphosphorofluoridate, as well as by physostigmine sulfate and -SH-blocking reagents, like HgCl-2 or 4-chloromercuribenzoic acid . o-Nitrophenol caused a competitive inhibition and phenetidine an uncompetitive inhibition.

Biochim Biophys Acta, 1975 May 6, 389(2), 345 - 57
Effect of lipid alkyl chain perturbations on the assembly of bacteriophage PM2; Cupp J et al.; The lipid-containing bacteriophage PM2 can produce infectious virus in cultures infected at temperatures up to 31.5 degrees C, but not at 34 degrees C . Its host, Pseudomonas BAL-31, grows at 34 degrees C and cultures infected at that temperature undergo lysis . Sucrose-gradient analysis shows that 34 degrees C lysates contain no PM2-like particles . Temperature-shift experiments establish that the thermally sensitive process is late in infection when virus assembly is taking place . Adamantanone, a small hydrophobic molecule that perturbs membrane hydrocarbon zones, prevents the production of infective virus . Concentrations which prevent virus production have no effect on host-cell growth or stability of mature virions . Adamantanone exerts its effects late in the infectious cycle, and lysates amde in its presence contain no PM2-like particles . These experiments, carried out at 25 degrees C, indicate that adamantanone prevents the assembly of stable PM2 virus . Spin-label studies suggest that the lipid alkyl chains of the host-cell membrane are in an "ordered" state at temperatures below about 33 degrees C and undergo a transition to a "disordered" state above that temperature . Furthermore, the addition of adamantanone perturbs the hydrocarbon zones, producing a greater degree of disorder even below 25 degrees C . Our findings suggest that the cell membrane can function and grow with the lipid alkyl chains in either the "ordered" or "disordered" state, but that the "ordered" state must be maintanined for PM2 assembly to occur.

J Clin Microbiol, 1975 May, 1(5), 462 - 4
Photoreactivation of Pseudomonas cepacia after ultraviolet exposure: a potential source of contamination in ultraviolet-treated waters; Carson LA et al.; Cells of a naturally occurring strain of Pseudomonas cepacia grown in distilled water were exposed to ultraviolet radiation . Irradiated samples incubated on membrane filters or in fluid recovery media in the absence of light showed no evidence of dark repair mechanisms . When samples were exposed to fluorescent light ranging from 50 to 950 foot candles (538 to 10,222 lux) of illumination, apparent photo-induced repair of ultraviolet injury resulted in 1- to 4-log increases in viable cells recovered.

J Bacteriol, 1975 May, 122(2), 557 - 64
Two distinct isocitrate lyases from a pseudomonas species; Bellion E et al.; The isocitrate lyases of acetate- and methylamine-grown Pseudomonas MA (Shaw strain) were studied . They were shown to be different by a variety of physical criteria including chromatographic elution patterns, heat inactivation kinetics, pH variation of Km values, and migration on polyacrylamide gels . The implications and significance of the existence of two enzymes in relation to the role of isocitrate lyase in methylamine utilization is discussed.

J Infect Dis, 1975 May, 131(5), 588 - 91
Bacteriophages and endotoxin in licensed live-virus vaccines; Moody EE et al.; In confirmation of recent reports, coliphages were found in seven of 19 unselected samples of the currently licensed live-virus vaccines . Coliphages and pseudomonas phage were found in 11 and 14, respectively, of the 20 bovine sera commonly used in the cell culture phase of virus vaccine production . The same lots of vaccine and serum were examined by the limulus assay for endotoxin, another product of bacterial contamination . Eighteen of 20 sera had detectable endotoxin-like activity . Our preliminary results suggest that endotoxin activity may serve as a sensitive indicator of residual products of previous bacterial contamination, including bacteriophages.

J Bacteriol, 1975 May, 122(2), 650 - 9
Interaction of Mg-2+ with peptidoglycan and its relation to the prevention of lysis of a marine pseudomonad; Rayman MK et al.; Intact cells of marine pseudomonad B-16 (ATCC 19855) which have been washed with a solution of NaCl require only 0.001 M MgSO4 and 100 to 300 times this concentration of NaCl or KCl to prevent lysis . Conversion of intact cells to mureinoplasts, a process involving removal of the outer double-track layer (outer membrane) and the periplasmic space layer of the cell wall, approximately doubled the requirement for the three salts to prevent lysis . The formation of protoplasts from mureinoplasts by removing the peptidoglycan layer again doubled the requirement for Na+ and K+ salts but increased the requirement for the Mg-2+ salt 200- to 300-fold . Cells of the marine pseudomonad suspended in solutions containing Mg-2+ salts failed to lyse on subsequent repeated suspension in distilled water, whereas cells presuspended in NaCl lysed immediately . Isolated envelope layers including the peptidoglycan layer, when dialyzed against solutiions containing Mg-2+ salts, retained Mg-2+ after subsequent suspension in distilled water . Envelope layers exposed to solutions of Na+ or K+ salts failed to retain these ions after exposure to distilled water . Na+ displaced Mg-2+ from the cell envelope layers . The results obtained indicate that the capacity of Mg-2+ salts at very low concentration to prevent lysis of intact cells and mureinoplasts of this organism is due primarily to the interaction of Mg-2+ with the peptidoglycan layer of the cell wall . Ion interaction with the layers lying outside of the peptidoglycan layer contributes only a small amount to the mechanical strength of the wall.

Biochem J, 1975 May, 147(2), 327 - 34
The purification and properties of L-histidine--2-oxoglutarate aminotransferase from Pseudomonas testosteroni; Hacking AJ et al.; 1 . Inducible L-histidine--2-oxoglutarate aminotransferase was purified some 170-fold from extracts of Pseudomonas testosteroni . 2 . The preparation showed only one major component after electrophoresis on polyacrylamide gels, though additional minor bands were observed when samples concentrated on a DEAE-cellulose column were used . 3 . The molecular weight of the enzyme was found to be approx . 70000 by chromatography on Sephadex G-200 . 4 . The purification scheme produced enzyme that was inactive in the absence of pyridoxal 5'-phosphate . 5 . The equilibrium constant for the reaction L-histidine+2-oxoglutarate equilibrium imidazolylpyruvate+L-glutamate was 0.49 . 6 . The reaction mechanism was Ping Pong . 7 . The enzyme was shown to have only low activity towards aromatic amino acids and was highly specific for 2-oxoglutarate.

Mikrobiologiia, 1975 May-Jun, 44(3), 552 - 4
{NAD-dependent N-methylglutamate dehydrogenase--new enzyme metabolizing methylamine in methylotrophs}; Netrusov AI; The properties of N-methylglutamate dehydrogenase, a new enzyme of methylamine metabolism in a facultative methylotroph Pseudomonas methylica, strain 2, were investigated . The enzyme was strictly dependent on the NAD presence and was not connected with particles in cell-free preparations . The enzyme appeared to be incucible and its activity increased after the growth of the cells in a medium containing methylamine.

Biochim Biophys Acta, 1975 Apr 29, 386(2), 590 - 602
Subunit structure of D-galactose dehydrogenase from Pseudomonas saccharophila; Wengenmayer F et al.; 1 . D-Galactose dehydrogenase from Pseudomonas saccharophila (molecular weight 102 000) dissociates in 8 M urea into its subunits (molecular weight 25 000) which migrate in polyacrylamide gels, containing 8 M urea, as a single band . 2 . The N-terminal residue determination by the dansyl method revealed only serine . 3 . The C-terminal group determination with carboxypeptidase A and B indicated the sequence -Tyr-His-Leu . Leucine as the single C-terminal amino acid was confirmed by the tritiation method and by tritiation and subsequent degradation with carboxypeptidases . 4 . The fragmentation of D-galactose dehydrogenase (24 mol methionine per mol enzyme) by CNBr resulted in six peptides, as detected in disc electrophoresis and substantiated by end group determination, indicating the identity of the subunits . 5 . The treatment of D-galactose dehydrogenase (24 mol lysine and 52 mol arginine per mol enzyme) with trypsin and subsequent peptide mapping showed 21, perhaps 22 peptides, indicating a structure comprising four identical subunits.

Biochim Biophys Acta, 1975 Apr 14, 387(1), 115 - 28
The kinetics and specificity of electron transfer from cytochromes and copper proteins to P700; Wood PM et al.; The rates of electron transfer to P700 from plastocyanin and cytochrome f have been compared with those from three other c-type cytochromes and azurin, a copper protein resembling plastocyanin . Three different disruptive techniques were used to expose P700; digitonin, Triton X-100 and sonication . The following rate constants were measured at 25 degrees C, pH 7.0, with digitonin-treated chloroplasts: plastocyanin, 8 x 10(7)M(-1) x s(-1); red-algal cytochrome c-553, 1.9 x 10(7)M(-1) x s (-1); Pseudomonas cytochrome c-551, 8 x 10(6)M(-1) x s (-1); azurin, less than or = 3 x 10(5)M(-1) x s (-1); cytochrome f, less than or = 2 x 10(4)M(-1) x s (-1); mammalian cytochrome c, less than or = 2 x 10(4)M(-1) x s (-1) . For electron transfer from plastocyanin, the effects of ionic strength, pH and temperature were also studied, and saturation effects found in earlier work were avoided by a full consideration of the various secondary reactions and inclusion of superoxide dismutase . The relative rates are discussed in relation to photosynthetic electron transport.

J Clin Microbiol, 1975 Apr, 1(4), 339 - 44
Cultural and biochemical characteristics and fatty acid composition of Pseudomonas diminuta and Pseudomonas vesiculare; Kaltenbach CM et al.; The cultural characteristics, biochemical activity, and cellular fatty acid composition of Pseudomonas diminuta and Pseudomonas vesiculare provided means for differentiation of these closely related species . Broth cultures of P . diminuta showed turbid growth and a distinct surface pellicle after 24 h at 35 C . P . vesiculare had no pellicle, and only light, diffuse growth was observed . All strains of P . vesiculare oxidized maltose and hydrolyzed esculin to varying degrees; P . diminuta was negative in these tests . The cellular fatty acids of these two species were similar, except that P . diminuta possessed a C19 cyclopropane acid which was not detected in P . vesiculare.

J Gen Microbiol, 1975 Apr, 87(2), 260 - 72
Isolation of an inducible amidase from Pseudomonas acidovorans AE1; Alt J et al.; A bacterial strain, AEI, which hydrolysed acetanilide, was isolated from soil and identified as Pseudomonas acidovorans . Numerous amides, esters and enzyme inhibitors were tested as amidase inducers . Phenacetin was chosen as inducer for the large scale cultivation of these organisms because it was less toxic to the bacteria than acetanilide . The induction increased the enzymic activity 250-fold . In comparison, the type culture strain of P . acidovorans, ATTCCI5668, had no amidase activity which could be induced by phenacetin . Optimal growth conditions were established with respect to the concentration of carbon source and inducer so that about 10% of the extractable bacterial protein consisted of the amidase . The organisms were lysed with lysozyme in the presence of EDTA and the enzyme was isolated mainly by column chromatography procedures . A preparation form 60 g (wet wt) bacteria yielded about 100 mg highly purified amidase with a specific activity of 137 mugmol substrate hydrolysed/min/mg protien . In addition to acetanilide, the purified enzyme hydrolysed several other amides and esters . As standard substrate, p-nitroacetanilide was chosen.

Am J Cardiol, 1975 Apr, 35(4), 481 - 6
Hemodynamic consequences of total removal of the tricuspid valve without prosthetic replacement; Robin E et al.; Tricuspid valvulectomy without prosthetic replacement has been advocated as a life-saving measure in the treatment of Pseudomonas endocarditis of the tricuspid valve . This report describes the hemodynamic data obtained in 10 patients before and after removal of the tricuspid valve . Seven patients remained free of carciac decompensation, but right heart failure developed in three . Analysis of the preoperative data did not permit differentiation of these two groups of patients.

Can J Microbiol, 1975 Apr, 21(4), 577 - 9
Dissimilation of the lignin model compound veratrylglycerol-beta-(o-methoxyphenyl) ether by Pseudomonas acidovorans: initial transformations; Crawford RL et al.; The initial transformadomonas acidovorans are demethylation of the 4-methoxyl group of the 3,4-dimethoxyphenyl portion of the molecule, and a NAD+-linked dehydrogenation of the benzyl alcohol group . Based on these findings and those described before, an overall degradation scheme is postulated.

Proc Soc Exp Biol Med, 1975 Apr, 148(4), 981 - 5
Correlation of the critical micelle concentrations of surfactants with their effects on a bacterial demethylase; Pinto JT et al.; The activity of a sarcosine dehydrogenase isolated from a strain of Pseudomonas is enhanced by the addition of Triton X-100, Brij 35, and Tween 80, and is inhibited by deoxycholate and Sarkosyl NL-97 . 2,6-Dichlorophenolindophenol, which is used as the oxidant in the dehydrogenase assay, has also been employed as an indicator in the spectrophotometric determination of the critical micelle concentrations (CMC) of both the nonionic and anionic detergents under conditions optimal for the enzyme analyses . A correlation between the activation or inhibitory activities of the surfactants and their CMC values has been established.

Biochim Biophys Acta, 1975 Mar 28, 384(1), 235 - 41
Isolation of an extracellular neutral proteinase from Pseudomonas fragi; Porzio MA et al.; A single proteolytic enzyme (EC 3.4.4.-) was isolated from culture supernatants of Pseudomonas fragi with 20% yielded and 60-fold purification by means of stepwise DEAE-Sephadex batch adsorption, ammonium sulfate precipitation, gel filtration and DEAE-cellulose chromatography . The enzyme was Zn-2+ activated and Ca-2+ stabilized, had optimum activity at pH 6.5--8.0 and 40 degrees C . The molecular weight range was 40 000--50 000 as determined by dodecylsulfate gel electrophoresis, gel filtration and Zn assay . This proteinase has properties similar to other extracellular bacterial neutral proteinases.

Arch Microbiol, 1975 Mar 12, 103(1), 71 - 6
Enzymatic analysis of the pathways of glucose catabolism and gluconeogenesis in Pseudomonas citronellolis; O'Brien RW; Extracts of Pseudomonas citronellolis cells grown on glucose or gluconate possessed all the enzymes of the Entner-Doudoroff pathway . Gluconokinase and either or both 6-phosphogluconate dehydratase and KDPG aldolase were induced by growth on these substrates . Glucose and gluconate dehydrogenases and 6-phosphofructokinase were not detected . Thus catabolism of glucose proceeds via an inducible Entner-Doudoroff pathway . Metabolism of glyceraldehyde 3-phosphate apparently proceeded via glyceraldehyde 3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, enolase and pyruvate kinase . These same enzymes plus triose phosphate isomerase were present in lactate-grown cells indicating that synthesis of triose phosphates from gluconeogenic substrates also occurs via this pathway . Extracts of lactate grown-cells possessed fructose diphosphatase and phosphohexoisomerase but apparently lacked fructose diphosphate aldolase thus indicating either the presence of an aldolase with unusual properties or requirements or an alternative pathway for the conversion of triose phosphate to fructose disphosphate . Cells contained two species of glyceraldehyde 3-phosphate dehydrogenase, one an NAD-dependent enzyme which predominated when the organism was grown on glycolytic substrates and the other, an NADP-dependent enzyme which predominated when the organism was grown on gluconeogenic substrates.

Arch Microbiol, 1975 Mar 10, 102(3), 253 - 60
Comparative biochemistry of alpha-glucan-utilization in Pseudomonas amyloderamosa and Pseudomonas saccharophila: physiological significance of variations in the pathway; Norrman J et al.; Growth patterns on and utilization of various alpha-glucans were investigated in Pseudomonas amyloderamosa and P . saccharophila . Maltose, maltodextrins (average chain length 7 glycosyl units) and glycogen supported excellent growth of both organisms and were extensively metabolized, although glycogen utilization in P . saccharophila was preceded by a prolonged lag phase . P . amyloderamosa produced limited growth on amylopectin and the carbohydrate was only partly degraded . It seemed likely that many of the unit chains liberated from amylopectin had a length exceeding the substrate range accepted by the maltodextrin permease (transport) system . A correlation was established between the pH of the medium and the utilization of glycogen and amylopectin for growth in P . amyloderamosa . The carbohydrates were at least partly utilizable at pH 6.0, whereas they could not support any growth at pH 6.5 . Most likely, the lack of growth at the higher pH reflected the low activity of isolamylase at this pH . The enzyme patterns of maltodextrin catabolism in the two bacteria were established . Intracellularly, maltodextrin phosphorylase and 4-alpha-glucanotransferase occurred in both . Degradation of extracellular alpha-glucans was mediated by a mainly intracellular isoamylase in P . amyloderamosa, whereas P . saccharophila possessed an extracellular alpha-amylase and a firmly cell-bound pullulanase.

Cancer Chemother Rep, 1975 Mar-Apr, 59(2 Pt 1), 411 - 9
Intensive cyclophosphamide (NSC-26271) therapy for solid tumors; Mullins GM et al.; Cyclophospamide was given in two dose schedules to 25 patients with a variety of nonlymphoid solid tumors . Eleven patients were given 18 courses of cyclophosphamide at a total dose of 60 mg/kg . Sixteen patients received 26 courses at a total dose of 100 mg/kg . Two patients were treated with both regimens . Partial responses were achieved in two patients treated with 60-mg/kg dose of cyclophosphamide . One of these patients had osteogenic sarcoma and the other had renal carcinoma . The higher dose also produced two partial responses, one in a patient with anaplastic carcinom a of the lung and the other in a patient with anaplastic carcinoma of the lung and the other in a patient with embryonal testicular carcinoma . Mean leukocyte counts fell to a nadir of 1400 cells/mm after 60 mg/kg while they dropped to below 1000 cells/mm for 5 days after 100 mg/kg of cyclophosphamide . Mean platelet counts remained above 150,000 platelets/mm after both cyclophosphamide schedules . In fective complications were documented aftter three of the 18 courses at 60 mg/kg and after ten of the 26 courses at 100 mg5kg . In the latter group, there were three episodes of bacteremia, including one death from pseudomonas sepsis . Nonhematologic toxicity noted with the 100-mg/kg dose of cyclophosphamide included rare instances of electrocardiogram changes and serum enzyme alterations compatible with myocardial toxicity . The intensive cyclophosphamide therapy did not appear to result in an increased antitumor response in malignancies usually considered to be refractory to alkylating agents.

Proc Natl Acad Sci U S A, 1975 Mar, 72(3), 849 - 53
Intermediates in the biosynthesis of double-stranded ribonucleic acids of bacteriophage phi 6; Coplin DL et al.; Pseudomonas phaseolicola infected with bacteriophage phi 6 synthesized all three viral double-stranded RNA segments, three single-stranded RNAs, and three replicative intermediate-like RNAs in the presence of rifampin . The single-stranded RNA intermediates sedimented and electrophoresed along with melted viral double-stranded RNA, annealed with melted viral double-stranded RNA, and were transient in nature . The relative amounts of the single-stranded RNA intermediates varied during the infection cycle and were altered in the presence of chloramphenicol . The replicative intermediate-like RNAs sedimented faster than double-stranded RNA, failed to enter 2.5% polyacrylamide gels, eluted with double-stranded RNA from a CF-11 cellulose column, were precipitated with single-stranded RNA in 2 M LiC1, and yielded three genome-size pieces of double-stranded RNA upon digestion with RNase . These results are consistent with the hypothesis that complementary strands of the phi 6 double-stranded RNAs are synthesized asynchronously during the infection cycle.

J Thorac Cardiovasc Surg, 1975 Mar, 69(3), 377 - 81
Localization of endovascular infection by selective catheterization with serial cultures; Bach MC et al.; Pseudomonas infection developed at the suture line of an aortic graft in a patient 13 years after the operation . The site of the infection was localized by quantitative blood cultures taken with the aid of selective arterial catheterization . This technique may be of great help in localizing the source of endovascular infection in difficult cases.

J Bacteriol, 1975 Mar, 121(3), 933 - 41
Purification and properties of a serine protease from Pseudomonas matophilia; Boethling RS; The extracellular protease of Pseudomonas maltophilia was partially purified by ammonium sulfate precipitation and chromatography on Sephadex G-75 and Bio-rex 70 . Gel electrophoresis revealed minor impurities . The enzyme exhibited the following properties: (i) molecular weight, 35,000; (ii) A see article; 10.8; (iii) isoelectric point, 9.3; (iv) pH optimum, 10.0; (v)s20, w equal 3.47 . The enzyme was rapidly inactivated by ethylenediaminetetracetate, but activity could be partially restored with divalent cations . Of those tested, Ca2+, Sr2+, Ba2+, Co2+, Cu2+, Mg2+, and Zn2+ were all effective . Both phenylmethylsulfonylfluoride and diisopropylfluorophosphate were powerful inhibitors of protease activity, but L-1-tosylamide-2-phenylethylchloromethyl ketone, iodoacetic acid, and iodoacetamide were without effect . The enzyme hydrolyzed the esters N-acetyl-L-tyrosine ethyl ester and alpha-N-benzoyl-L-arginine ethyl ester (BAEE) with Km values of 10.4 and 3.4 mM, respectively . The hydrolysis of BAEE was also inhibited by phenylarsonic acids . The kinetics of inhibition by m-nitrophenylarsonate were of the mixed type, and the K1 was 1.8 mM . The data followed a theoretical curve for a 1:1 enzyme-inhibitor complex with a dissociation constant of 1.8 mM . Inhibition by m-nitrophenylarsonate was pH dependent and followed a theoretical curve for the titration of a protonated group with a pKa of 7.0.

Arch Neurol, 1975 Mar, 32(3), 204 - 5
Neurological sequelae of malignant external otitis; Faden A; The neurological sequelae of malignant external otitis (MEO) form a characteristic syndrome . Following Pseudomonas external otitis, usually in an elderly, diabetic patient, either isolated facial nerve paralysis or multiple cranial nerve palsies develop . Once extensive neurological signs have developed, recovery rarely occurs . We saw a patient with MEO and multiple cranial nerve palsies who recovered following an extended course of gentamicin sulfate and carbenicillin disodium therapy.

Appl Microbiol, 1975 Mar, 29(3), 382 - 7
Bacterial metabolism of arylsulfonates: role of meta cleavage in benzene sulfonate oxidation by Pseudomonas testosteroni; Ripin MJ et al.; Pseudomonas testosteroni H-8 oxidizes certain lower alkylbenzene sulfonates at rates inversely related to the length of the alkyl group . Appreciable Q(O)2 values were observed for benzene sulfonate (BS), toluene sulfonate (TS), and ethylbenzene sulfonate (EBS), but not for propylbenzene sulfonate (PS) and higher homologues . Catechol oxidation was catalyzed by a constitutive catechol-2,3-oxygenase (EC 1.99.2.a) . Yellow meta cleavage products accumulated when BS-grown cells were exposed to catechol, 4-methylcatechol, 3-methylcatechol, EBS and PS, but not BS or TS . Traces of a yellow metabolite (probably 2-hydroxymuconic semialdehyde) were detectable during growth on BS . PS completely inhibited growth on BS, but not on L-leucine or nutrient broth . Also, PS antagonized respiration on BS and catechol, but not glutamate, the extent of inhibition being directly related to PS concentration . Formation of a meta cleavage product from PS, and inhibition of catechol oxidation by PS, suggested that the actual inhibitor may not be PS itself, but a metabolite.

Prikl Biokhim Mikrobiol, 1975 Mar-Apr, 11(2), 195 - 202
{Study of inosine transformation into 5'-inosinic acid by the culture of Pseudomonas trifoli}; Kazarinova LA et al.; The transformation of inosine into 5'-inosine acid by Pseudomonas trifolii cells was studied . The synthesis of 5'-inosine acid can be performed by both live intact and dry cells . The effectiveness of inosine phosphorylation depends on the ratio of the inosine and phosphate donor concentrations and the amount of cells . The temperature and pH effect on activity of nucleoside phosphotransferase, phosphomonoesterase and 5'-nucleotidase has been studied . The influence of surface active substances and metal ions on the synthesis of 5'-inosine acid has been investigated . Optimal conditions for the inosine transformation by the above culture have been established.

J Biol Chem, 1975 Feb 10, 250(3), 905 - 11
Sequential degradation of keratan sulfate by bacterial enzymes and purification of a sulfatase in the enzymatic system; Nakazawa K et al.; Pseudomonas sp . IFO-13309 and Actinobacillus sp . IFO-13310, bacteria which exhibit a symbiotic growth in a medium containing keratin sulfate as a sole carbon source, were isolated from soil . Extracts of these organisms were shown to contain an endoglycosidase, a sulfatase, and exo-beta-D-galactosidase, and an exo-beta-D-N-acetylglucosaminidase which, together, catalyze an extensive cleavage of corneal keratan sulfate . The Pseudomonas extract was particularly rich in the endoglycosidase activity and poor in the exoglycosidase activities . The Actinobacillus extract, in sharp contrast, contained principally the exoglycosidases . The sulfatase activity did not show this marked difference in distribution . A sulfatase was purified from the crude extract of Actinobacillus . The purified sulfatase reacted little or not at all with keratan sulfate, but acted on 2-acetamido-2-deoxy-6-O-sulfo-D-glucose, 2-acetamido-2-deoxy-6-O-sulfo-beta-D-glucosyl-(1 leads to 3)-D-galactose, and a tetrasaccharide trisulfate having 2-acetamido-2-deoxy-6-O-sulfo-D-glucose at the nonreducing end (prepared from keratan sulfate with an endogalactosidase) . The enzyme removed one sulfate group from the tetrasaccharide trisulfate, producing an oligosaccharide which, unlike the parent oligosaccharide, was susceptible to hydrolysis with exo-beta-D-N-acetylglucosaminidase . The data suggest that the nonreducing end is the only site at wich enzymatic desulfation is carried out.

J Biol Chem, 1975 Feb 10, 250(3), 912 - 7
Purification of Keratan Sulfate-endogalactosidase and its action on keratan sulfates of different origin; Nakazawa K et al.; A glycosidase which attacks corneal keratan sulfate was purified from extracts of Pseudomonas sp . IFO-13309 . When corneal keratan sulfate was degraded by the purified enzyme, Sephadex G-50 chromatography indicated the presence of a number of oligosaccharides differing in size and sulfate content . The characterization of two major fractions of the oligosaccharides indicated that the point of enzyme attack is limited to the endo-beta-D-galactoside bonds in which nonsulfated D-galactose residues participate . The enzyme, unlike ordinary exo-beta-D-galactosidases, did not catalyze the hydrolysis of phenyl beta-D-galactoside . Moreover, beta-D-galactosyl-(1 leads to 3)-2-acetamido-2-deoxy-beta-D-glucosyl-(1 leads to 3)-beta-D-galactosyl-(1 leads to 4)-D-glucose ("lacto-N-tetraose") was completely refractory to the action of this enzyme, suggesting that a structure of the type, X-(1 leads to 3)-beta-D-galactosyl-(1 leads to 4)-Y, is not the only specificity-determining factor, i.e . neighboring sugars, X and Y, or even larger portions of substrate molecule must have an important effect . Compared with corneal keratan sulfate, keratan sulfates from human nucleus pulposus and shark cartilage were attacked at lower rates with a resultant production of oligosaccharides of relatively large size . The result is in agreement with the view that considerable variations exist in the structure of keratan sulfates of different origin, and further suggests that the enzyme may serve as a useful reagent in studying these variations.

Biochem J, 1975 Feb, 145(2), 305 - 9
Studies on the subunit structure of 4-deoxy-5-oxoglucarate hydro-lyase (decarboxylating) from Pseudomonas acidovorans; Jeffcoat R; 1 . Homogeneous preparations of D-4-deoxy-5-oxoglutarate hydro lyase (decarboxylating)(EC4.2.1.41) were analysed in the ultracentrifuge by the high-speed sedimentation-equilibrium method of Yphantis (1964) . The molecular weight in 0.1 M-potassium phosphate buffer, pH 7.2, in 6M-guanidine hydrochloride and in 0.1 M-beta-mercaptoethanol in 6M-guanidine hydrochloride was 113,000, 56,000 and 30,400 respectively . Polyacrylamidegel electrophoresis in the presence of sodium dodecyl sulphate indicated a minimum molecular weight of 30,500 . 2 . Measurement of the thiol content of the enzyme, before and after reduction with NaBH4 or dithiothreitol under denaturing conditions, indicated the presence of eight thiol residues and two interchain disulphide bridges/enzyme molecule . 3 . Amino acid analysis showed that the intact enzyme contains a total of approximately 100 arginine and lysine residues, but digestion of the enzyme with trypsin yielded about 49 peptides staining with ninhydrin in a peptide "map" . 4 . With the knowledge that the enzyme contains only two substrate-binding sites, it is suggested that the enzyme probably consists of four polypeptide chains arranged in an alpha2beta2 confirmation.

J Clin Pathol, 1975 Feb, 28(2), 149 - 55
Strains of Pseudomonas putrefaciens from clinical material; Holmes B et al.; Eight strains of Pseudomonas putrefaciens have been received from among 466 strains of Pseudomonas submitted to the Computer Trials Laboratory for identification over the last eight years . Two of the strains of P . putrefaciens from patients with otitis media and otitis externa respectively appear to have played a pathogenic role . The biochemical characteristics of these eight strains were compared with those of seven culture collection strains.

Can J Microbiol, 1975 Feb, 21(2), 173 - 80
A mixed bacterial population in a continuous culture with and without kaolinite; Maigetter RZ et al.; Chromobacterium lividum and a Pseudomonas sp . were grown in pure and mixed continuous culture with and without the clay-mineral, kaolinite . Irrespective of the growth conditions, C . lividum adhered to the wall of the culture vessel whereas the Pseudomonas sp . showed no such tendency, at least visually . During mixed culture studies, the organism which was initially established in the culture dominated . The ratio between C . lividum and the Pseudomonas sp . was about 20:1 when C . lividum was first established and 1:2 when the Pseudomonas sp . was first grown . The indirect fluorescent antibody technique provided a rapid method for differentiating the mixed cultures when the bacterial concentration was sufficient for microscopic analysis . During both pure and mixed continuous culture studies, the addition of kaolinite reduced the C . lividum but not the Pseudomonas sp . population.

Br J Exp Pathol, 1975 Feb, 56(1), 34 - 43
Protective properties and haemagglutinins in serum from humans and in serum from mice injected with a new polyvalent Pseudomonas vaccine; Jones RJ et al.; Mice given single injections of a polyvalent pseudomonas vaccine produced anti-pseudomonas haemagglutinins against the 16 component immunogens of the multivalent vaccine . Mice passively immunized with sera from vaccinated mice were protected against lethal challenge by 8/10 strains of Ps . aeruginosa of homologous serotype . Protection by the serum was inversely proportional to the virulence of the challenge strains . Anti-pseudomonas haemagglutinins were always present in sera which passively protected mice against pseudomonas infection . Low levels of anti-pseudomonas haemagglutinins were present in some sera which failed to passively immunize mice against pseudomonas infection . Anti-pseudomonas haemagglutinins and antibodies involved in passive protection were mainly in the IgM fractions of mouse serum . Control human sera contained anti-pseudomonas haemagglutinins against most serotypes of Ps . aeruginosa . Sera from patients with burns contained high levels of anti-pseudomonas haemagglutinins against some but not all serotypes of Ps . aeruginosa . Sera from both controls and patients with burns passively protected mice against pseudomonas infection.

Hoppe Seylers Z Physiol Chem, 1975 Feb, 356(2), 193 - 201
Studies on a 2,3-diaminopropionate: ammonia-lyase from a Pseudomonad; Vijayalakshmi KR et al.; 2,3-Diaminopropionate:ammonia-lyase, an induced enzyme in a Pseudomonas isolate, has been purified 40-fold and found to be homogeneous by disc gel electrophoresis and by ultracentrifugation . Some of its properties have been studied . The optimum pH and temperature for activity are 8 and 40 degrees C, respectively . The enzyme shows a high degree of substrate specificity, acting only on 2,3-diaminopropionate; the D-isomer is only one-eighth as effective as the L-form . L-Homoserine and DL-cystathionine are not substrates, and 3-cyanolalanine does not inhibit its activity . It is a pyridoxal phosphate enzyme which requires free enzyme sulphhydryls for activity . The Km values for L-2,3-diaminopropionate and pyridoxal phosphate are 1mM and 25 muM, respectively . The molecular weight of the enzyme is about 80 000 as determined by gel filtration . On treatment with 0.5M urea or guanidine by hydrochloride, the enzyme dissociates into inactive subunits with an approximate molecular weight of 45 000 . One mole of the active enzyme binds one mole of pyridoxal phosphate . The bacterial enzyme seems to be quite different in many of its properties from the rat liver enzyme which also exhibits the substrate specificity of cystathionine gamma-lyase.

J Bacteriol, 1975 Feb, 121(2), 695 - 9
Cyanide production from glycine by a homogenate from a Pseudomonas species; Wissing F; A cell-free preparation with cyanide-producing activity was obtained from a bacterium, strain C, of the genus Pseudomonas . To preserve activity, an oxidizing agent, e.g., phenazine methosulphage (PMS), had to be added to the cell suspension before disruption by sonic treatment . By the procedure described, a total homogenate made from a 15% (wet weight) bacterial suspension in tris(hydroxymethyl)aminomethane-hydrochloride buffer (0.05 M, pH 8.2) and with PMS (0.4mM) exhibited about 8% of the activity obtained from a suspension of untreated bacteria . In the presence of flavine-adenine dinucleotide (0.3 mM) and PMS (0.4mM), the activity was augmented to about 16% of that of the intact cells . By gradient centrifugation the homogenate was separated into three fractions . The main enzyme activity was associated with those fractions which by electron microscopy were found to consist of membranous structures.

Biochemistry, 1975 Jan 28, 14(2), 300 - 7
DNA-dependent RNA polymerase from Pseudomonas BAL-31 . II . Transcription of the allomorphic forms of bacteriophage PM2 DNA; Zimmer SG et al.; Transcription of the supercoiled form (I) and the relaxed circular form (II) of bacteriophage PM2 DNA was studied utilizing the DNA-dependent RNA polymerase from its host, Pseudomonas BAL-31 . Transcription of both templates is continuous for up to 2 hr, but proceeds at a two-fold higher rate on I than on II . This difference is mainly due to a 2.2-fold higher rate of chain initiation on I . When rifampicin (Rif) is added ater 10 min of synthesis, (1) transcription of II ceases by 30 min with a maximum product length of 7000 nucleotides (number average) being produced; (2) transcription of I continues with little rate reduction and with the product reaching 16,000 nucleotides (number average) by 2 hr . Sucrose gradient analysis shows that the product of II achieves maximum size 20 min after Rif addition and sediments in three peaks of 24, 33, and 39 S (approximately one-third, two-thirds, and one genome lengths) . The product of I has a heterogeneous distribution and grows continuously with a large fraction reacting greater than 3 genome lengths by 90 min . The same differences in synthesis kinetics, Rif inhibition, and product size distribution are observed when I and II are transcribed by Escherichia coli RNA polymerase . These experiments show that (i) PM2 form I DNA is transcribed mainly by a process of continuous chain elongation, with little chain termination occurring; (ii) PM2 form II is transcribed by a process of continuous chain initiation, elongation, and termination of yield discrete products . Thus, the tertiary structure of circular DNA influences chain termination by RNA polymerase.

Biochemistry, 1975 Jan 28, 14(2), 290 - 9
DNA-dependent RNA polymerase from Pseudomonas BAL-31 . I . Purification and properties of the enzyme; Zimmer SG et al.; The DNA-dependent RNA polymerase from Pseudomonas BAL-31, the host for bacteriophage PM2, has been purified 154-fold using differential centrifugation, chromatography on DEAE-cellulose, ammonium sulfate precipitation, and sucrose gradient centrifugations at low and high ionic strength . The resulting enzyme is free of enzyme activities which could interfere with transcription studies and is greater than 85% pure as judged by polyacrylamide gel electrophoresis . Like other bacterial RNA polymerases, its subunit structure is beta'beta sigma alpha2 . From gel electrophoresis the beta', beta, and alpha subunits have approximately the same molecular weights as those from Escherichia coli, whereas the sigma subunit is 5% larger (89,000 vs . 85,000) . A summation of the subunits yields a molecular weight of 485,000 for the holoenzyme . Like other bacterial RNA polymerases, it sediments as a monomer (15 S) at low ionic strength (0.065) and as a dimer (22 S) at high ionic strength (0.75) . Its activity is stimulated three-fold by monovalent cations (K+,NH4+, NA+) with additional stimulation provided by divalent cations (Mg2plus, Mn2plus) . The transcription of phage PM2 form I (supercoiled) DNA has an ionic strength optimum of 0.26 for continuous long-term synthesis, and over an ionic strength range of 0.09-0.46 "plateau-type" kinetics are not observed . The sigma subunit is required for optimal PM2 transcription . The enzyme is sensitive to the same inhibitors of transcription as the RNA polymerase from E . coli, it has a temperature optimum of 28 degrees, and it is 50% inactivated by heating 10 min at 41 degrees . It has template preference similar to E . coli polymerase and shows little preference for homologous templates . With various DNAs the order of template activities is T7 greater than PM2 I congruent to T4 greater than PM2 II (relaxed circular form) greater than lambda-c greater than calf thymus greater than BAL-31 DNA . Phage PM2 form I DNA is transcribed at a twofold greater rate than PM2 form II DNA by this enzyme.

Eur J Biochem, 1975 Jan 15, 50(3), 603 - 9
3Alpha-hydroxysteroid dehydrogenase from Pseudomonas testosteroni: kinetic properties with NAD and its thionicotinamide analogue; Skalhegg BA; The kinetics of 3alpha-hydroxysteroid : NAD oxidoreductase (EC 1.1.1.50) from Pseudomonas testosteroni (ATCC 11996) have been investigated . The kinetic analysis based on initial activity measurements and product inhibition studies, indicates that the addition of substrate to the enzyme and the release of products from it, follows an obligatory order (ordered bi bi mechanism) . The ability of the enzyme to utilize the thionicotinamide analogue of NAD (sNAD) as cofactor has been investigated using various 3alpha-hydroxysteroids from both the C19, C21, and C24 series . The results show that the reaction velocity with sNAD as the cofactor is generally lower than with NAD . The decrease, however, varies considerably, being negligible with some steroids such as litocholic acid and deoxycholic acid and very pronounced with other such as tetrahydrocortisol and tetrahydrocortisone . The introduction of an 11beta-hydroxy or an 11-oxo group into the steroid molecule significantly reduces the ability of the enzyme to attack the 3alpha-hydroxy group . No such effect could be seen when the 11-hydroxy group was in the alpha-position . The results also indicate that, whereas NAD can serve as cofactor for both the monomeric and the dimeric forms of the enzyme, sNAD only acts as cofactor for the monomeric form . Thus sNAD is a valuable tool for the study of the reversible, concentration-depenedent monomeric-dimeric transition of the 3alpha-hydroxysteroid dehydrogenase.

J Biol Chem, 1975 Jan 10, 250(1), 276 - 80
Concentration-dependent association of delta5-3-ketosteroid isomerase of Pseudomonas testosteroni; Benson AM et al.; Gel chromatography and ultracentrifugation studies show that delta5-3-ketosteroid isomerase of Pseudomonas testosteroni a dimer with a molecular weight of 26,800 at concentrations below 1 mg per ml, undergoes reversible, concentration-dependent association at higher enzyme concentrations . In the concentration range between 0.04 and 15.6 mg per ml, apparent molecular radii of 23 A to 36 A and molecular weights of 26,000 to 69,000 were observed . The latter value represents the weight average molecular weight of two or more ploymerization species in rapid equilibrium, rather than a discrete polymeric form of the enzyme . The isomerase dimer has been found to be unusually stable to dissociation upon dilution, even at concentrations in the nanogram per ml range . Evidence is presented which suggests that the enzyme is present as a dimer in P . testosteroni cells and that this is a catalytically active species . The isomerase monomer has been obtained and its molecular weight studied by gel electrophoresis in the presence of sodium dodecyl sulfate . A new determination of the extinction coefficient of the isomerase gives the value of 0.336 for the absorbance at 280 nm in a 1-cm light path of a solution containing 1 mg of the isomerase per ml.

J Biol Chem, 1975 Jan 10, 250(1), 271 - 5
Effect of protein concentration on the molecular weight of delta5-3-ketosteroid isomerase; Tivol WF et al.; The molecular weight of delta-5-3-ketosteroid isomerase from Pseudomonas testosteroni was determined by means of sedimentation equilibrium and exclusion chromatography over a wide range of enzyme concentrations in 0.2 M potassium phosphate buffer, pH 7.0 . In addition, the sedimentation constant of the enzyme was determinded over an extended range of concentrations . The enzyme was found to have a molecular weight of 26,000 plus or equal to 1,000, suggesting that it is a dimer of identical or similar 13,400 molecular weight polypeptide chains . In the ultracentrifuge this dimeric species was found to undergo aggregation at enzyme concentrations above 2 mg per ml and dissociation at enzyme concentrations below 0.05 mg per ml . Exclusion chromatography studies indicate that under the conditions of chromatography the oligomeric enzyme is partially dissociated at enzyme concentrations in the range 0.2 to 0.002 mug per ml . These results suggest that under conditions of enzyme assay in 0.2 M potassium phosphate buffer, pH 7.0, isomerase is in a monomeric state of aggregation.

J Biol Chem, 1975 Jan 10, 250(1), 344 - 7
Radiocopper in L-tryptophan 2,3-dioxygenase isolated from Pseudomonas acidovorans grown in the presence of 64Cu (II); Brady FO; L-Tryptophan 2,3-dioxygenase (EC 1.13.11.11), isolated from L-tryptophan-induced Pseudomonas acidovorans, ATCC 11299b, which has been grown in a medium containing 64Cu(NO3)2, has been shown to contain radiocopper . At several stages of purification of the enzyme samples were taken, and these were subjected to disc acrylamide gel electrophoresis in the presence of 10 mM L-tryptophan . After electrophoresis the position of the yellow heme band, corresponding to tryptophan oxygenase, was visually located, and the gels were sliced and counted . A large peak of radioactivity was seen to occur at the location on the gel of tryptophan oxygenase no matter what the stage of purification . Treatment of each sample before electrophoresis for 30 min at 37 degrees with gamma-globulins prepared from rabbits sensitized to homogeneous pseudomonad tryptophan oxygenase greatly reduced this peak of radioactivity, whereas treatment of each sample with rabbit preimmune gamma-globulin did not . This direct demonstration of the presence of coper in pseudomonad tryptophan oxygenase, using 64-Cu, avoided the problems and artifacts inherent in the usual techniques of copper analysis and unequivocally refutes the recent contention of Ishimura and Hayaishi ((1973) J . Biol.Chem . 248, 8610-8612) "that copper is not an essential component of L-tryptophan 2,3-dioxygenase of Pseudomonas." The presence of copper in pseudomonad and rat liver tryptophan oxygenases, previously reported by us (Brady, F . O., Monaco, M . E., Forman, H . J., Schutz, G., and Feigelson, P . (P . (1972) J . Biol . Chem . 247, 7915-7922), is reaffirmed by the experiments reported herein.

Antibiotiki, 1975, 20(12), 1077 - 81
{New antibiotically active fluoroglucide from Pseudomonas aurantiaca}; Esipov SE et al.; A new metabolite with an antibiotic activity against grampositive bacteria in concentrations of 0.1--1.0 gamma/ml was isolated from the culture fluid of Pseudomonas aurantiaca . The study of its physico-chemical properties showed that it was di-2,4-diacetylfluoroglucylmethan . The conclusion was confirmed by synthetic studies . Di-2,4-diacetylfluoroglucylmethan belongs to the group of the antibiotics, derivatives of fluoroglucin, characteristics metabolites of Pseudomonas aurantiaca.

Langenbecks Arch Chir, 1975, Suppl, 169 - 72
{Treatment of pseudomonas infection following burn injuries with specific antitoxic Ig-therapy}; Stadtler K et al.; The mortality rate of mice after standardized Pseudomonas infection was increased by preinjection of a sublethal dose of burn toxin . This effect was eliminated by a specific antitoxic Ig . After a sublethal burn followed by infection, 60% of the animals died, while treatment with specific Ig reduced the mortality rate to 3.3% . This might be a new way to improve the treatment of severely burned patients.

Int J Pept Protein Res, 1975, 7(4), 335 - 9
On the 3alpha-hydroxysteroid dehydrogenase from Pseudomonas testosteroni . The effect of denaturing agents; Skalhegg BA; Highly purified preparations of the 3alpha-hydroxysteroid:NAD- oxidoreductase (E.C.1.1.1.50) from Pseudomonas testosteroni (ATCC 11996) which consist of two major isoenzymes, with traces of a third, have been split into two enzymatically inactive polypeptides A and B by the use of sodium dodecylsulphate, urea and guanidinium hydrochloride . Both polypeptides have a molecular weight of 25,000 +/- 2,500 as shown by thin-layer gel chromatography and ultracentrifugations . They differ, however, in charge as shown by electrophoresis on cellulose acetate strips in the presence of 8 M urea . Each of the isoenzymes, have molecular weight of about 50,000 and thus consist of two subunits . The presence of the three isoenzymes may be explained by the following combinations of the subunits, AA, AB and BB . Close to 100% of the original activity towards the three substrates, androsterone tetrahydrocortisone and desoxycholate could be restored within 24 h when the inactivated enzyme was diluted in order to remove the effect of the denaturant.

Int J Vitam Nutr Res, 1975, 45(2), 144 - 52
Effects of pyridoxal phosphate N-oxide and 2'-hydroxy pyridoxal phosphate on L-aspartate beta-decarboxylase; Shibatani T et al.; Pyridoxal phosphate N-oxide and 2'-hydroxypyridoxal phosphate served as the coenzyme for aspartate beta-decarboxylase (EC 4.1.1.12) from Pseudomonas dacunhae . Reconstituted enzymes with those pyridoxal phosphate analogues exhibited an absorption band near 370 nm . Close to 1 mole of vitamin B6 derivative is bound per minimal catalytic unit with high affinity . The decarboxylase, desulfinase, and transaminase activites of the both pyridoxal phosphate derivate-enzymes are relatively low . But the Km values for aspartate and cysteine sulfinate are not affected.

Acta Biochim Pol, 1975, 22(1), 57 - 66
Molecular properties of the inducible lupanine hydroxylase from growing cultures of Pseudomonas lupanini; Rogozinski J; An improved method for isolation of lupanine hydroxylase, giving a 450-fold purification, is presented . The molecular weight of the enzyme is about 72 000, and the sedimentation coefficient S20, W 5.05 . The enzyme contains a component similar to Pseudomonas cytochromes c . Its oxidation-reduction potential was found to be below + 45 mV.

J Bacteriol, 1975 Jan, 121(1), 160 - 4
Kinetics of Na+-dependent K+ ion transport in a marine pseudomonad; Hassan HM et al.; The effect of external Na plus concentration on the transport of K plus was studied using K plus-depleted cells of a marine pseudomonad . K plus transport was found to be a saturable process and requires Na plus . The initial rates for K plus transport over a range of external K plus concentrations were measured in suspensions containing various fixed concentrations of Na plus . Reciprocals of the initial rates for K plus transport were plotted against reciprocals of the external concentration of K plus or Na plus to yield two primary Lineweaver-Burk plots . The experimental data were found to fit bisubstrate enzyme kinetics, with a sequential type mechanism . However, the initial rate data did not allow distinction between ordered or random mechanisms . The results suggest that Na plus and K plus form a ternary complex with a specific K plus carrier molecule on the outer surface of the membrane prior to translocation and the release of K plus inside the cell.

Can J Microbiol, 1975 Jan, 21(1), 43 - 50
Kinetics of Naplus-dependent amino acid transport using cells and membrane vesicles of a marine pseudomonad; Sprott GD et al.; Sodium ion is required for transport of L-alanine and alpha-aminoisobutyric acid (AIB) into cells and membrane vesicles of a marine pseudomonad . Initial rate data obtained at various Naplus and amino acid concentrations were tested for fit by least squares analysis to sequential and Ping-Pong equations . The sequential case is preferred statistically at the 99% confidence limit . Cotransport of AIB and Naplus in cells could not be detected, perhaps explained by efflux of Naplus shown to occur from these cells . The kinetic analysis is consistent with formation of a ternary complex involving Naplus and the amino acid.

Antonie Van Leeuwenhoek, 1975, 41(4), 569 - 74
Characteristics of weak H2S-producing isolates of Pseudomonas putrefaciens from human infections; Levin RE; Isolates from human clinical specimens which produced salmon-colored colonies were found to grow at 2C and produce ornthine decarboxylase, DNase, trimethylamine and only slight amounts of H2S . The notably weak ability of such isolates to produce H2S was originally regarded as a negative reaction . The DNA base composition of such isolates was found to range from 48.3 to 49.2 moles % GC . These observations, along with bacteriocin typing, resulted in identification of the isolates as Pseudomonas putrefaciens.

Antonie Van Leeuwenhoek, 1975, 41(2), 135 - 46
Substrate inhibition in Pseudomonas oxalaticus OX1: a kinetic study of growth inhibition by oxalate and formate using extended cultures; Dijkhuizen L et al.; Pseudomonas oxalaticus OX1 has been grown in a mineral salts medium with oxalate or formate as the sole source of carbon and energy . At concentrations of these substrates above 50 mM inhibition of growth was indicated by a long and variable lag phase in batch culture . This inhibition was further studied by estimating maximum specific growth rates at different substrate concentrations using the extended culture technique for control of the substrate concentration . With formate, inhibition became apparent at substrate concentrations above 20 mM, whereas oxalate inhibited growth at concentrations above 15 mM . Complete inhibition was not observed even at concentrations of 100 mM . A number of inhibition functions were fitted with the experimental data using computer analysis . The results indicated that the Haldane equation was the simplest function to describe quantitatively the kinetics of the observed substrate inhibition . Studies on the rate of oxygen uptake at different concentrations of oxalate indicated that respiration was much more sensitive to inhibition than growth . However with formate, inhibition of respiration was not observed up to concentrations of 50 mM, indicating that different mechanisms may underlie the observed growth inhibition by the two substrates.

Antonie Van Leeuwenhoek, 1975, 41(1), 97 - 100
Bacteriocin typing of Pseudomonas putrefaciens from food, human clinical specimens, and other sources; Williams JL et al.; With the use of 4 bacteriocin donor strains of Pseudomonas putrefaciens most low G+C strins were readily distinguished from high G+C strains . Two bacteriocin donor strains exhibited autoinhibition when subjected to bacteriocin typing.

Mikrobiologiia, 1975 Jan-Feb, 44(1), 55 - 60
{Growth and antibiotic formation of bacteria of genus Pseudomonas on media with n-alkanes of low molecular weight}; Kvasnikov EI et al.; The ability to assimilate n-alkanes form hexane to decane was studied among 495 collection strains and 27 freshly isolated strains belonging to the genus Pseudomonas . All freshly isolated strains and over one third of collection cultures of Ps . aurantiaca grow on mineral media with n-alkanes of low molecular weight, but do not assimilate heavy paraffins . The strains of Ps . aeruginosa, Ps . fluorescens and Ps . putida, isolated from oilbearing soils, and individual collection cultures, belonging to the two latter species, can assimilate both n-alkanes of low molecular weight (C6--C10) and heavy paraffins . Contrary to Ps . aurantiaca, other species of the Pseudomonas genus lose the ability to assimilate n-alkanes of low molecular weight after cultivation on rich organic media . An increase in the concentration of the mixture of low molecular weight paraffins (to 20 per cent by volume) has no toxic effect on the Pseudomonas bacteria whose biomass has a high content of protein and all necessary amino acids . The strains of Ps . aurantiaca produce a highly active antibiotic preparation consisting of floroglucine derivatives on the defined medium with n-alkanes of low molecular weight . The ratio between components of the preparation obtained on the media with n-alkanes and on the optimal organic media is different.

Antonie Van Leeuwenhoek, 1975, 41(1), 89 - 95
A novel inducible formaldehyde dehyrogenase of Pseudomonas sp . (RJ1); Mehta RJ; A novel, pyridine-nucleotide-independent, inducible formaldehyde dehydrogenase acitivity was detected in cells of Pseudomonas sp . (RJ1) propagated on methylamine and oxalated . The pH optimum of the dehydrogenase was 7.0 . Dichlorophenol-indophenol or potassium ferricyanide served as an electron acceptor . The rate of reduction of these electron acceptors was shown to be stimulated by phenazine methosulfate . The dehydrogenase was inhibited by parahydroxymercuric benzoate and iodoacetamide . This inhibition suggests that the enzyme contains sulfhydryl groups . The stoichiometry of the reaction in terms of oxygen uptake to formate formation was 0.5, which agrees with the theoretical value.

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