Achromobacter

Proc Natl Acad Sci U S A, 1990 Nov, 87(22), 8960 - 4
Posttranslationally processed structure of the human platelet protein smg p21B: evidence for geranylgeranylation and carboxyl methylation of the C-terminal cysteine; Kawata M et al.; smg p21A and -B are small GTP-binding proteins that share putative effector and consensus C-terminal sequences with ras p21 proteins . In the present report, we showed that human platelet smg p21B became labeled when intact platelets were incubated with exogenous {3H}mevalonolactone and when a purified preparation of smg p21B was incubated with bovine brain membranes and S-adenosyl-L-{methyl-3H}methionine . In addition, we demonstrated by gas chromatography/mass spectrometry that treatment of smg p21B with Raney nickel released a geranylgeranyl moiety in a molar ratio of about 1:1 . In contrast, treatment of smg p21B with NH2OH or KOH yielded no evidence for the presence of a palmitoyl thioester . Extensive digestion of smg p21B with Achromobacter protease I yielded two C-terminal tripeptides that contained serine and cysteine in a molar ratio of 2:1 . Both peptides were modified by a thioether-linked geranylgeranyl group . One of the peptides comigrated with a 3H-labeled proteolytic product of methylated smg p21B on reverse-phase HPLC and this peptide appeared at the same retention time as that of the other peptide after being treated with KOH . Since the cDNA-predicted C-terminal sequence of smg p21B contains a unique Ser-Ser-Cys peptide within its C-terminal domain, -Lys-Lys-Ser-Ser-Cys-Gln-Leu-Leu184, these results indicate that smg p21B is posttranslationally modified by geranylgeranylation of Cys-181 and suggest that further modifications cause proteolytic removal of the three predicted C-terminal amino acids followed by partial methylation of the cysteinyl carboxyl group.

J Biochem (Tokyo), 1990 Nov, 108(5), 816 - 21
Cloning and sequence analysis of cDNA for Trimeresurus flavoviridis phospholipase A2, and consequent revision of the amino acid sequence; Oda N et al.; A cDNA clone of Trimeresurus flavoviridis phospholipase A2 was isolated from the venom gland cDNA library using DNA amplified by polymerase chain reaction with oligonucleotide primers corresponding to the N- and C-terminal sequences of this enzyme . The amino acid sequence deduced from the cDNA nucleotide sequence determined by the dideoxy termination method was found to be inconsistent in the segment comprising the 69th to 81st positions from that reported previously {Tanaka, S . et al . (1986) J . Biochem . 99, 281-289} . Reinvestigation of the amino acid sequence of the peptide covering the sequence in question showed that the amino acid sequence predicted from the nucleotide sequence is correct . The sequence contains Asn-Asn-Gly (positions 69-71) and it was found that the Asn-Gly bond easily undergoes alpha-beta transpeptidation when digested with Achromobacter protease I at pH 9.0 but not seriously at pH 6.8 . It is likely that the transpeptidation reaction caused a failure in the previous sequence determination . The cDNA clone obtained was 597 base pairs long and contained an open reading frame of 414 base pairs coding for 138 amino acid residues . A typical signal peptide sequence (16 amino acid residues long), located at the N-terminal moiety of the deduced sequence, was immediately followed by a polypeptide which corresponds to the mature enzyme . Northern blot analysis showed a single transcript only in the poly(A)+ RNA fraction of the venom gland but not in those of many other organs tested.

J Biol Chem, 1990 Oct 25, 265(30), 18345 - 50
Identification of lysine 134 in the steroid-binding site of the sex steroid-binding protein of human plasma; Namkung PC et al.; The sex steroid-binding protein of human plasma SBP (or sex hormone-binding globulin, SHBG) was specifically inhibited with the alkylating affinity label, 17 beta-{{( 2-14C}bromoacetyl)oxy}-5 alpha-androstan-3-one . The natural ligand, 5 alpha-dihydrotestosterone, was shown to protect against inactivation and labeling . The steroid-binding activity of the protein was abolished when approximately 1 mol of label was incorporated into 1 mol of dimeric SBP . In order to identify and locate the labeled amino acid in the steroid-binding site, the steroidal portion of the bound label was first removed and the protein was digested with Achromobacter protease and subdigested with trypsin . Seven radioactive peptides were isolated, sequenced, and found to contain the common sequence QVSGPLTSXR . Residue X was identified as lysine-134 from the SBP amino acid sequence (Walsh, K . A., Titani, K., Kumar, S., Hayes, R., and Petra, P . H . (1986) Biochemistry 25, 7584-7590) . The results indicate that only 1 of the 2 lysine-134 residues in the homodimer was labeled . This suggests that the steroid-binding site is constructed from an association of the two subunits in an AB to BA "sandwich" configuration with lysine-134 residue of one subunit on one surface near the D-ring and the lysine-134 of the other subunit at the opposite end of the steroid, or well away from the steroid-binding site . Although the nature of the data does not allow description of a specific role for lysine-134, its proximity to the 17 beta-OH of the steroid nucleus suggests participation in the binding process through direct or indirect hydrogen bonding.

Experientia, 1990 Oct 15, 46(10), 1066 - 8
Inhibitory effect of halocyamine, an antimicrobial substance from ascidian hemocytes, on the growth of fish viruses and marine bacteria; Azumi K et al.; Halocyamine A, an antimicrobial substance isolated from hemocytes of the solitary ascidian Halocynthia roretzi, inhibited in vitro the growth of fish RNA viruses (infectious hematopoietic necrosis virus and infectious pancreatic necrosis virus) . Pretreatment of RNA virus with halocyamine A reduced the infectivity of the virus toward host cells . The growth of marine bacteria, Achromobacter aquamarinus and Pseudomonas perfectomarinus, was also inhibited by halocyamine A but that of Alteromonas putrefaciens and Vibrio anguillarum was not . These results suggest that halocyamine may have a role in the defense mechanisms of H . roretzi against marine viruses and bacteria.

J Biochem (Tokyo), 1990 Oct, 108(4), 604 - 8
Purification and complex formation analysis of a cysteine proteinase inhibitor (cystatin) from seeds of Wisteria floribunda; Hirashiki I et al.; Seeds of Wisteria floribunda contain several kinds of cysteine proteinase inhibitor (cystatin) . We purified and characterized one of these inhibitors, named WCPI-3 . The molecular weight of WCPI-3 was estimated to be 17,500 and 15,700 by gel filtration and SDS-PAGE, respectively . The isoelectric point was 5.7 . WCPI-3 formed an equimolar complex with native papain and the dissociation constant was estimated to be 6.1 nM . Complex formation between WCPI-3 and Cys25-modified papain, such as S-carboxy-methylated or S-carbamoylmethylated papain, could not be observed by gel filtration or native PAGE analysis . A peptide fragment derived from WCPI-3 digested by Achromobacter proteinase (lysyl endopeptidase) had the amino acid sequence of VVAGVNYRFVLK . The VVAG sequence in this fragment corresponds to the conserved sequence QVVAG which is considered to be one of binding regions to cysteine proteinases . The amino acid sequence of the amino-terminal portion (34 residues) of WCPI-3 was highly homologous to that of oryzacystatin from rice seeds.

J Mol Recognit, 1990 Oct-Dec, 3(5-6), 181 - 6
Enzymatic semisynthesis of human insulin: an update; Morihara K; Peptide bond formation can be enzymatically catalysed by the reverse reaction of proteases . Application is seen in the industrial production of human insulin . Human insulin derivative can be enzymatically prepared using either porcine insulin or the single chain B(1-29)-A(1-21) insulin precursor as the starting material . This is accomplished by either (1) digesting the starting material at Lys29 with Achromobacter lyticus protease I (Ach) and then coupling with Thr-X (X = blocking residue) (two-step reaction) or (2) subjecting Ala-B30 of porcine insulin or Gly-A1 of the single chain insulin precursor to transpeptidation with Thr-X (one-step reaction) . Trypsin and Ach can be used for either type of reaction and, in the immobilized form, for the two-step reaction . Since the single chain insulin precursor can be produced by gene technology (yeast), use of immobilized trypsin or Ach and the two-step reaction using the single chain insulin precursor as the starting material ensures the continuous production of human insulin making it a feasible method for industrial manufacture.

J Bacteriol, 1990 Sep, 172(9), 4945 - 50
Phosphorylation of the VirG protein of Agrobacterium tumefaciens by the autophosphorylated VirA protein: essential role in biological activity of VirG; Jin SG et al.; Agrobacterium tumefaciens virulence genes are induced by plant signals through the VirA-VirG two-component regulatory system . The VirA protein is a membrane-spanning sensor molecule that possesses an autophosphorylating activity, and the VirG protein is a sequence-specific DNA-binding protein . In this report, we demonstrate that the VirG protein is phosphorylated by the VirA protein and that the phosphate is directly transferred from the phosphorylated VirA molecule (phosphohistidine) to the VirG protein . The chemical stability of the phospho-VirG bond suggested that the VirG protein was phosphorylated at the aspartate and/or glutamate residue . The phosphorylated VirG protein was reduced with tritiated sodium borohydride and subjected to proteolytic digestion with the Achromobacter protease I enzyme . The resulting peptide fragments were separated by C8 reversed-phase high-pressure liquid chromatography, and the tritium-labeled peptide was sequenced . Amino acid sequence data showed that the aspartate residue at position 52 was the only site phosphorylated . Changing this aspartate into asparagine resulted in a nonphosphorylatable and biologically nonfunctional gene product . As a control, a randomly chosen aspartate was changed into an asparagine (position 72), and no effect on its phosphorylation or biological activity was observed . Unlike its homologs, including CheA-CheY, EnvZ-OmpR, and NtrB-NtrC, the phospho-VirG molecule was very stable in vitro . The possible implications of these observations and the function of VirG phosphorylation in vir gene activation are discussed.

Bioorg Khim, 1990 Aug, 16(8), 1040 - 4
{Site-specific endonucleases LplI and AagI}; Sokolov NN et al.; New site-specific endonucleases LplI and AagI have been isolated from the Lactobacillus plantarum and Achromobacter agile cells, respectively . The enzymes' purification stages included treatment of cell-free extracts with polyethylenimine, fractionation in two-phase system by Albertsson's method, chromatography on blue Sepharose and DEAE-cellulose . The results of cleavage of a 5'-32P-labelled oligodeoxynucleotide duplex by restriction endonucleases LplI and AagI indicate that these enzymes recognize and cut the sequence AT decreases CGAT, being therefore true isoschizomers of the ClaI restriction endonuclease from Caryophanon latum . The L . plantarum strain has 400 fold endonuclease productivity as compared with the ClaI producent and is perspective for preparative isolation of LplI.

J Biochem (Tokyo), 1990 Jul, 108(1), 139 - 43
Amino acid sequence of a lectin from Japanese frog (Rana japonica) eggs; Kamiya Y et al.; The complete amino acid sequence and the location of disulfide bonds of a lectin from Japanese frog (Rana japonica) eggs, which specifically agglutinates transformed cells, are presented . The sequence was determined by analysis of peptides generated by digestion of the S-carboxyamidomethylated protein with Achromobacter protease I, or chymotrypsin, and by chemical cleavage with BNPS-skatole or cyanogen bromide . The lectin is a single-chain protein consisting of 111 residues, with a pyroglutamyl residue at the amino terminus . Four disulfide bonds link half-cystinyl residue 19 to 72, 34 to 82, 52 to 97, and 94 to 111 . The sequence and the location of the disulfide bonds are highly homologous to those of bull frog (Rana catesbeiana) egg S-lectin . They are also homologous to human angiogenin, a tumor angiogenesis factor, and a family of pancreatic ribonucleases.

J Biochem (Tokyo), 1990 Jul, 108(1), 133 - 8
Complete amino acid sequence of rat kidney ornithine aminotransferase: identity with liver ornithine aminotransferase; Oyama R et al.; The complete amino acid sequence of rat kidney ornithine aminotransferase {EC 2.6.1.13} is presented . The 404-residue sequence was determined by analysis of peptides generated by digestion of the S-carboxyamidomethylated protein with CNBr, Achromobacter protease I, arginylendopeptidase, or Staphylococcus aureus V8 protease . Mueckler and Pitot have reported the amino acid sequence of the rat liver enzyme (440 residues) as predicted from the nucleotide sequence of the cDNA {Mueckler, M.M . & Pitot, H.C . (1985) J . Biol . Chem . 260, 12993-12997} . The amino acid sequence of the rat kidney enzyme presented herein coincides with residue 36 (Gly) through 440 (Phe) of the predicted precursor protein, indicating that the liver and kidney enzymes are identical, and that the enzyme is processed at the amino-terminal region after translation.

Biochem Biophys Res Commun, 1990 Jun 29, 169(3), 1235 - 41
EPR spectra of ferric cytochromes c' from five strains of Achromobacter xylosoxidans at low temperature and their temperature dependence; Yoshimura T et al.; The EPR spectra at low temperature (6 K) and their temperature dependence (10-93 K) for five ferric cytochromes c' isolated from chemoheterotrophic bacteria, Achromobacter xylosoxidans NCIB 11015 (formerly Alcaligenes sp . NCIB 11015), GIFU 543, GIFU 1048, GIFU 1051, and GIFU 1764 are reported . The EPR spectral results indicate that the ground state of the heme iron(III) of cytochromes c' from these chemoheterotrophic bacteria can appear to be in an admixed spin state which consists of predominant S = 5/2 with a slight S = 3/2 character . The EPR spectra were compared with those for ferric cytochromes c' from photosynthetic bacteria and the other ferric hemoproteins.

Blood, 1990 Jun 15, 75(12), 2349 - 56
Rapid purification and characterization of human platelet glycoprotein V: the amino acid sequence contains leucine-rich repetitive modules as in glycoprotein Ib; Shimomura T et al.; Glycoprotein V (GPV) is a membrane-associated, 82 Kd platelet glycoprotein that is hydrolyzed during thrombin activation to yield 69 Kd fragment . We have developed a rapid and simple method for isolation of the protein from platelet extracts using a combination of gel permeation, anion-exchange, and lectin affinity chromatography . The partial amino acid sequence was determined by analysis of peptides generated by digestion of the S-carboxyamido-methylated protein with Achromobacter protease I or cyanogen bromide . The sequence shows a remarkable periodicity of leucine residues, which is homologous to the consensus sequence of a highly diversified protein super-family with a common repetitive module . Thrombin cleavage site was determined to be located at the C-terminal region of GPV by analysis of the products separated by sizing and reversed-phase high performance liquid chromatography . By lectin blot analysis, the existence of mucin-type carbohydrate chains was indicated, as well as the existence of asparagine-linked carbohydrate chains shown by the amino acid sequence analysis . From these data, we report a structural model of GPV that is analogous to glycoprotein Ib.

J Biol Chem, 1990 Jun 5, 265(16), 9188 - 93
A distinct human testis and brain mu-class glutathione S-transferase . Molecular cloning and characterization of a form present even in individuals lacking hepatic type mu isoenzymes; Campbell E et al.; mu-Class glutathione S-transferases (GSTs) were identified in all 13 human testes and 28 brains examined; even subjects whose livers were devoid of mu-GSTs expressed extrahepatic GSTs of this class . Testes and brains from individuals with mu-class GSTs in their livers had additional forms that also reflected the liver phenotypes . An isoenzyme with an isoelectric point of 5.2, which was a major GST in testis and present as well in cerebral cortex but not detected in any livers, was identified and purified . Sequence analysis of peptides derived by cleavage of the testicular mu-class GST by Achromobacter protease I revealed distinct aspects of primary structure not found previously in any mammalian mu-class GSTs . These unique features included a blocked and extended amino terminus and 3 additional residues (Pro-Val-Cys) at the carboxyl terminus . This structure was confirmed by molecular cloning and sequencing of cDNAs derived from human testis and brain libraries . In the coding region the mRNA of the brain-testis mu-class GST was 75% homologous with that of the liver form, and its 3'-untranslated sequence was mostly divergent, indicating that it is the product of a separate gene . Distinct catalytic and structural properties of the testis-brain mu-class GSTs suggest that these GSTs may be uniquely involved in blood-barrier functions common to both organs.

J Appl Bacteriol, 1990 May, 68(5), 495 - 504
Numerical analysis of electrophoretic protein patterns of 'Achromobacter' group B, E and F strains from human blood; Holmes B et al.; Thirty-two clinical strains representing 'Achromobacter' groups B, E and F were characterized by one-dimensional SDS-PAGE of cellular proteins . All the strains were isolated from blood samples from hospital patients in the United Kingdom . The protein patterns, which contained 40 to 45 discrete bands, were highly reproducible and were used as the basis for a numerical analysis which included all the protein bands . The 32 'Achromobacter' strains formed two clusters at the 77% S level . The first, phenon 1, included the 28 group B and the two group E strains and the second, phenon 2, contained the two strains of group F . The strains in each phenon were characterized by a clearly distinct pattern of protein bands . Phenon 1 could be further divided at the 87% S level into three subphenons which correlated with differences in the principal bands found between 40.0 and 42.5 kD . Strains of group E clustered with group B strains from which they could not be distinguished by protein patterns . We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides a useful method for the classification of this group of bacteria . Reference strains of each of the PAGE types identified are available from NCTC for inclusion in future studies.

Electrophoresis, 1990 May, 11(5), 382 - 91
Numerical analysis of sodium dodecyl sulphate-polyacrylamide gel electrophoretic protein patterns for the classification, identification and typing of medically important bacteria; Costas M; A numerical analysis of one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoretic protein patterns of three separate bacterial groups is described . Similarity between patterns was estimated using a correlation coefficient and is represented diagramatically by the unweighted pair-group with arithmetic averages method of clustering . Protein patterns were scanned using a laser densitometer interfaced directly to a microcomputer, where calculations were performed semi-automatically with specially written software . A method for the normalization of patterns from different gels is utilized . Whole cell protein patterns clearly distinguished strains representing various groups of Achromobacter . Similarly the two biovars of the Group EF-4 bacteria could be separated after analysis of partial "background patterns" as could the different species of Providencia . In addition, whole patterns could be used to type the various strains of P . rettgeri . These examples serve to illustrate the power, versatility and potential of the technique when applied to problems of classification, identification and typing of bacteria of medical interest . Its advantages and disadvantages, when compared to other available electrophoretic methodologies, are discussed, as is the future application of such technology to the hospital laboratory.

Biol Chem Hoppe Seyler, 1990 May, 371(5), 441 - 6
Susceptibility of baboon aorta elastin to proteolysis; Desfontaines L et al.; Elastin was purified from baboon aorta using Achromobacter collagenase and its susceptibility to proteolysis by various enzymes was studied . Human leukocyte elastase (HLE) hydrolysed baboon aortic elastin 8 times faster than human cathepsin G . Bovine chymotrypsin had virtually no activity against this substrate . The kinetic constants V and {S50} of aortic elastin hydrolysis by HLE (0.15 microM) were 0.00286 mg x ml-1 x min-1 and 0.158 mg x ml-1, respectively . One mg of this elastin could be saturated with 5.6 micrograms of HLE . As with elastins isolated from other sources, the hydrolysis of baboon aortic elastin by HLE was highly sensitive to ionic strength, and a biphasic effect was obtained with increasing NaCl concentrations . A nearly 2-fold stimulation of elastolysis was observed at a 0.15M NaCl concentration . Further increase in ionic strength led to a continuous decrease of the rate of elastolysis which paralleled the decrease of adsorption of elastase to baboon aortic elastin . Cathepsin G, but not bovine alpha-chymotrypsin, was able to stimulate the rate of hydrolysis of baboon aortic elastin by HLE . A 1.7 fold stimulation was observed for a 1:1 molar ratio of the two proteinases and rose to 2.1 for a HLE/Cat . G ratio equal to 8.

Diagn Microbiol Infect Dis, 1990 May-Jun, 13(3), 253 - 9
Antimicrobial activity of Ro 24-6778, a covalent bonding of desmethylfleroxacin and desacetylcefotaxime; Jones RN; A new "dual action" cephalosporin (Ro 24-6778), representing an ester-linked desacetylcefotaxime and desmethylfleroxacin was tested against 287 aerobic bacteria . Ro 24-6778 was found to be very active (minimal inhibitory concentrations {MIC90}, less than or equal to 0.5 micrograms/ml) against Enterobacteriaceae, Streptococcus spp., and Aeromonas hydrophila . Moderate Ro 24-6778 activity (MIC90, 1-8 micrograms/ml) was demonstrated against Bacillus spp., Staphylococcus spp . (including oxacillin-resistant strains), Flavobacterium spp., Enterococcus durans, and Acinetobacter anitratus . More Ro 24-6778-resistant strains (MIC90, 16- greater than 32 micrograms/ml) were usually found among the enterococci, Xanthomonas hydrophila, Pseudomonas spp., and Achromobacter xyloxidans isolates . These preliminary Ro 24-6778 MIC test results show a spectrum superior (93.4% of strains susceptible at less than or equal to 8 micrograms/ml) to the comparison drugs (cefotaxime or fleroxacin) and possible clinical utility for therapy of many fluoroquinolone- or cephalosporin-resistant strains.

J Biochem (Tokyo), 1990 Feb, 107(2), 292 - 7
Structure of recombinant human interleukin 5 produced by Chinese hamster ovary cells; Minamitake Y et al.; The complete peptide map of purified recombinant human interleukin 5 (rhIL-5) was determined to verify its primary structure, glycosylation sites, and disulfide bonding structure . Each peptide fragment generated by Achromobacter protease I (API) digestion was purified and characterized by amino acid analysis and amino acid sequence analysis . After digestion with API, we could identify all the peptides which were expected from human IL-5 cDNA sequence . The analyses of sulfhydryl content in rhIL-5 molecule and disulfide-containing peptide obtained from API digestion indicated that active form of rhIL-5 existed as an antiparallel dimer linked by two pairs of Cys-44 and Cys-86 . In addition, we concluded that Thr-3 and Asn-28 were glycosylated . The results indicate that primary structure of rhIL-5 is highly homogeneous and observed heterogeneity is due to the difference in the content of carbohydrate.

J Biochem (Tokyo), 1990 Feb, 107(2), 236 - 41
Purification, amino acid sequence, and some properties of rabbit kidney lysozyme; Ito Y et al.; The lysozyme (rabbit kidney lysozyme) from the homogenate of rabbit kidney (Japanese white) was purified by repeated cation-exchange chromatography on Bio-Rex 70 . The amino acid sequence was determined by automated gas-phase Edman degradation of the peptides obtained from the digestion of reduced and S-carboxymethylated rabbit lysozyme with Achromobacter protease I (lysyl endopeptidase) . The sequence thus determined was KIYERCELARTLKKLGLDGYKGVSLANWMCLAKWESSYNTRATNYNPGDKSTDYGIFQ INSRYWCNDGKTPRAVNACHIPCSDLLKDDITQAVACAKRVVSDPQGIRAWVAWRNHCQ NQDLTPYIRGCGV, indicating 25 amino acid substitutions from human lysozyme . The lytic activity of rabbit lysozyme against Micrococcus lysodeikticus at pH 7, ionic strength of 0.1, and 30 degrees C was found to be 190 and 60% of those of hen and human lysozymes, respectively . The lytic activity-pH profile of rabbit lysozyme was slightly different from those of hen and human lysozymes . While hen and human lysozymes had wide optimum activities at around pH 5.5-8.5, the optimum activity of rabbit lysozyme was at around pH 5.5-7.0 . The high proline content (five residues per molecule compared with two prolines per molecule in hen or human lysozyme) is one of the interesting features of rabbit lysozyme . The transition temperatures for the unfolding of rabbit, human, and hen lysozymes in 3 M guanidine hydrochloride at pH 5.5 were 51.2, 45.5, and 45.4 degrees C, respectively, indicating that rabbit lysozyme is stabler than the other two lysozymes . The high proline content may be responsible for the increased stability of rabbit lysozyme.

Biol Chem Hoppe Seyler, 1990 Feb, 371(2), 137 - 44
Collagenase activation of latent matrix-degrading proteinases from human plasma fibronectin; Imhoff JM et al.; Fibronectin contains two latent gelatinolytic enzymes, FN-gelatinase and FN-laminase that can be activated in the presence of Ca2+ from the purified cathepsin D-produced 190-kDa fibronectin fragment . The results of this work show that Achromobacter collagenase cleaves fibronectin and generates an active FN-gelatinase . In contrast to the cathepsin D digest, the collagenase digest directly exhibits gelatinolytic activity without additional activation . The gelatinolytic activity of the total collagenase digest can be inhibited by phenylmethanesulfonyl fluoride, a serine proteinase inhibitor and by pepstatin A, an aspartic-acid proteinase inhibitor . FN-laminase activity, when assayed with its synthetic substrate GPAGPR and also with laminin was revealed after separation of the collagenase digest of fibronectin on heparin Ultrogel . FN-gelatinase and FN-laminase activities were found in heparin unretained and heparin strongly retained fractions . These results have demonstrated that in contrast to cathepsin D, Achromobacter collagenase activates two matrix-degrading proteinases from fibronectin, FN-Gelatinase und FN-Laminase.

J Hosp Infect, 1990 Feb, 15(2), 149 - 55
Absence of role for plasmids in resistance to multiple disinfectants in three strains of bacteria; Nagai I et al.; Three chlorhexidine-resistant strains, two of Achromobacter xylosoxidans, one of Pseudomonas cepacia, were isolated from clinical sources . The three strains showed cross-resistance to benzethonium chloride, benzalkonium chloride and alkyldiaminoethylglycine . Resistance persisted after treatment of the strains with acridine orange (12.5 mg-25 mg l-1) . Conjugation to appropriate recipients of Escherichia coli was unsuccessful and plasmid DNA could not be detected . These results suggest that plasmids do not play a role in resistance to multiple disinfectants in the strains of bacteria studied.

Biochem Biophys Res Commun, 1990 Jan 30, 166(2), 729 - 35
Isolation of a high specific activity pink, monomeric nitrous oxide reductase from Achromobacter cycloclastes; Hulse CL et al.; Nitrous oxide reductase from Achromobacter cycloclastes has been purified to homogeneity under aerobic conditions via DEAE-cellulose, phenyl-Sepharose, hydroxyapatite, and Sephacryl S-200 chromatography . It consists of a single polypeptide of MW 72 kDa, and contains 3.8 +/- 0.1 copper atoms per molecule . The enzyme is pink as isolated, yet exhibits a specific activity (86 U/mg) that is ca . 40 times greater than that observed for other N2O reductases under similar conditions . Double integration of the anomalous EPR spectrum at 77K showed the presence of 2.0 +/- 0.1 spins per molecule, implying the presence of EPR-silent copper atoms and/or spin-coupled mixed-valent centers.

Toxicon, 1990, 28(1), 43 - 54
Purification and amino acid sequence of basic protein I, a lysine-49-phospholipase A2 with low activity, from the venom of Trimeresurus flavoviridis (Habu snake); Yoshizumi K et al.; A basic protein (pI 10.2), named basic protein I, was purified to homogeneity from the venom of Trimeresurus flavoviridis (Habu snake) after four chromatographic steps . The amino acid sequence of this protein was determined by sequencing the S-pyridylethylated derivative of the protein and its peptides produced by chemical (cyanogen bromide and formic acid) and enzymatic (chymotrypsin, Achromobacter protease I, and Staphylococcus aureus V8 protease) cleavages . The protein consisted of 122 amino acid residues and was similar in sequence to phospholipases A2 from the venoms of crotalid and viperid snakes . A most striking feature of this protein is that aspartic acid at the 49th position common in phospholipases A2 is replaced by lysine . When the protein acted on 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphorylcholine, oleic acid was preferentially released, indicating that the protein has phospholipase A2 activity . Its molar activity toward 1,2-dilauroyl-sn-glycero-3-phosphorylcholine, however, was 1.5% that of T . flavoviridis phospholipase A2 isolated previously . The fact that both affinity to Ca2+ and reactivity to p-bromophenacyl bromide of basic protein I are approximately one order of magnitude lower than those of T . flavoviridis phospholipase A2 might explain the low activity of basic protein I.

CLAO J, 1990 Jan-Mar, 16(1), 49 - 52
Microbial keratitis and corneal ulceration associated with therapeutic soft contact lenses; Kent HD et al.; We reviewed the records of 22 patients whose corneal ulcers were associated with therapeutic soft contact lens wear . The patients required hospitalization on the Cornea Service at Wills Eye Hospital between January 1, 1978 and September 1, 1988 . A majority of the ulcers were associated with pseudophakic or aphakic bullous keratopathy (9 of 22 cases; 41%); neurotrophic/exposure keratitis was the second most common diagnosis (7 of 22; 32%) . Most patients used topical antibiotics (15 of 22; 68%) and/or corticosteroids (13 of 22; 59%) . Cultures were positive in 15 of 22 cases (68%) . Gram-positive organisms were isolated in 60% the culture-positive cases (9 of 15) . Streptococcus was the most common organism isolated (6 of 15 culture positive-cases; 40%) . Gram-negative organisms were found in four of 15 culture-positive ulcers (27%) . There was only one Pseudomonas infection in the series . Uncommon organisms--including Candida, atypical mycobacteria, Achromobacter, Acinetobacter and Micrococcus--were isolated in five cases . Therapeutic soft contact lens wearers are at risk for developing corneal ulcers; most often these are caused by gram-positive bacteria, especially streptococci, and uncommon organisms.

Agric Biol Chem, 1990 Jan, 54(1), 131 - 7
Limited proteolysis of silkworm antitrypsin by several proteinases; Sasaki T et al.; Silkworm antitrypsin (sw-AT) isolated from larval hemolymph was limitedly digested by Achromobacter lysylendopeptidase, alpha-chymotrypsin, subtilisin BPN', subtilisin Carlsberg, papain, or Pseudomonas elastase . Each proteinase could cleave specific site(s) around the reactive site identified for the reaction of sw-AT and bovine trypsin . Among these proteinases, only subtilisin BPN' was inhibited by sw-AT, although weakly . By the cleavable amino acid sequence in sw-AT, it was suggested that whether or not these proteinases were inhibited by sw-AT did not solely depend on their substrate specificities . The susceptibility to the attack of proteinase should indicate that this region is exposed on the molecular surface . The amino acid sequence in the COOH-terminal region slightly away from the reactive site in sw-AT had homology with that in the corresponding region of the serine proteinase inhibitor (serpin) group.

J Biol Chem, 1989 Dec 5, 264(34), 20625 - 31
Cloning, nucleotide sequence, and expression of Achromobacter protease I gene; Ohara T et al.; Achromobacter protease I (API) is a lysine-specific serine protease which hydrolyzes specifically the lysyl peptide bond . A gene coding for API was cloned from Achromobacter lyticus M497-1 . Nucleotide sequence of the cloned DNA fragment revealed that the gene coded for a single polypeptide chain of 653 amino acids . The N-terminal 205 amino acids, including signal peptide and the threonine/serine-rich C-terminal 180 amino acids are flanking the 268 amino acid-mature protein which was identified by protein sequencing . Escherichia coli carrying a plasmid containing the cloned API gene overproduced and secreted a protein of Mr 50,000 (API') into the periplasm . This protein exhibited a distinct endopeptidase activity specific for lysyl bonds as well . The N-terminal amino acid sequence of API' was the same as mature API, suggesting that the enzyme retained the C-terminal extended peptide chain . The present experiments indicate that API, an extracellular protease produced by gram-negative bacteria, is synthesized in vivo as a precursor protein bearing long extended peptide chains at both N and C termini.

Cornea, 1989 Dec, 8(4), 267 - 9
Achromobacter xylosoxidans corneal ulcer in a therapeutic soft contact lens wearer; Fiscella R et al.; Achromobacter xylosoxidans is an opportunistic organism that is usually seen in immunocompromised or immunosuppressed patients . It is an aerobic gram-negative rod, often confused with other more commonly seen gram-negative bacteria such as Pseudomonas aeruginosa . The organism is usually sensitive to extended spectrum penicillins such as carbenicillin and usually resistant to aminoglycosides and first generation cephalosporins . We wish to describe a corneal ulcer from A . xylosoxidans that developed in a patient wearing a therapeutic soft contact lens . The patient did not have a preexisting microbial keratitis and was not receiving corticosteroid therapy.

Protein Seq Data Anal, 1989 Dec, 2(6), 441 - 4
Determination of partial amino acid sequence of lipoate acetyltransferase of rat pyruvate dehydrogenase complex; Matuda S et al.; The partial amino acid sequence of rat lipoate acetyltransferase was determined using the intact protein and the peptides derived from a digest with Achromobacter protease I . The results showed the amino-terminal sequence of the mature enzyme to be (N) Ser-Leu-Pro-Pro-His-Gln-Lys-Val-Pro-Leu-Pro-Ser- Leu-Ser-Pro-Thr-Met-Gln-Ala-Gly-Thr-Ile-Ala-Arg-Trp-Glu-Lys . In addition, the sequences of two possible lipoyl-binding sites in the subunit, which are very similar to each other, were established.

J Biol Buccale, 1989 Dec, 17(4), 269 - 74
Action of a bacterial Achromobacter collagenase on the soft carious dentine: an in vitro study with the scanning electron microscope; Goldberg M et al.; Samples of carious human teeth treated in vitro for 48-92 hours with a bacterial Achromobacter collagenase in solution in borate buffer at 33 degrees were examined with the scanning electron microscope . The enzyme was seen to destroy soft carious dentine but not sound layers of dentine beneath the lesion . This finding may have clinical implications.

S Afr Med J, 1989 Nov 18, 76(10), 571 - 3
Fatal neonatal meningitis and ventriculitis caused by multiresistant Achromobacter xylosoxidans . A case report; Manjra AI et al.; Meningitis and ventriculitis in a 6-day-old neonate caused by a Gram-negative glucose-non-fermenting organism, Achromobacter xylosoxidans, was resistant to most antibiotics except ceftazidime and imipenem . The organism became resistant after 28 days treatment with ceftazidime . When the infant was 7 weeks old, imipenem became available but, in spite of 3 days of intravenous treatment, the organism was still recovered from ventricular cerebrospinal fluid and the child died . This would appear to be only the second report of neonatal meningitis caused by this organism.

Biochem Biophys Res Commun, 1989 Nov 15, 164(3), 1366 - 72
Spectroscopic evidence for a copper-nitrosyl intermediate in nitrite reduction by blue copper-containing nitrite reductase; Suzuki S et al.; The reactions of nitrogen monoxide (NO) with the blue copper-containing nitrite reductases from Alcaligenes sp . NCIB 11015 and Achromobacter cycloclastes IAM 1013 were investigated spectroscopically . The electron paramagnetic resonance (EPR) signals of the blue coppers vanished in the presence of NO at 77 K, being fully restored by the removal of NO . The additions of NO to the enzyme solutions resulted in the substantial bleaching of the visible absorption bands at room temperature . The reactions were also completely reversible . These results suggest the formation of a cuprous nitrosyl complex (Cu+-NO+), which is likely the intermediate in the enzymatic nitrite reduction.

J Biochem (Tokyo), 1989 Oct, 106(4), 548 - 51
The primary structure of porcine liver acylamino acid-releasing enzyme deduced from cDNA sequences; Mitta M et al.; Two overlapping cloned cDNAs encoding the entire amino acid sequences of the subunits of acylamino acid-releasing enzyme (AARE) {EC 3.4.19.1} have been isolated from porcine liver cDNA lambda gt10 cDNA libraries and sequenced . Sequence analyses of the cDNA and several Achromobacter protease I-digested peptides of the purified protein revealed that porcine liver AARE consists of four identical subunits, and each comprising a single chain of 732 amino acids with acetylmethionine at the N-terminus.

J Bacteriol, 1989 Oct, 171(10), 5467 - 72
Molecular relationship of chromosomal genes encoding biphenyl/polychlorinated biphenyl catabolism: some soil bacteria possess a highly conserved bph operon; Furukawa K et al.; All the genes we examined that encoded biphenyl/polychlorinated biphenyl (PCB) degradation were chromosomal, unlike many other degradation-encoding genes, which are plasmid borne . The molecular relationship of genes coding for biphenyl/PCB catabolism in various biphenyl/PCB-degrading Pseudomonas, Achromobacter, Alcaligenes, Moraxella, and Arthrobacter strains was investigated . Among 15 strains tested, 5 Pseudomonas strains and one Alcaligenes strain possessed the bphABC gene cluster on the XhoI 7.2-kilobase fragment corresponding to that of Pseudomonas pseudoalcaligenes KF707 . More importantly, the restriction profiles of these XhoI 7.2-kilobase fragments containing bphABC genes were very similar, if not identical, despite the dissimilarity of the flanking chromosomal regions . Three other strains also possessed bphABC genes homologous with those of KF707, and five other strains showed weak or no significant genetic homology with bphABC of KF707 . The immunological cross-reactivity of 2,3-dihydroxybiphenyl dioxygenases from various strains corresponded well to the DNA homology . On the other hand, the bphC gene of another PCB-degrading strain, Pseudomonas paucimobilis Q1, lacked genetic as well as immunological homology with any of the other 15 biphenyl/PCB degraders tested . The existence of the nearly identical chromosomal genes among various strains may suggest that a segment containing the bphABC genes has a mechanism for transferring the gene from one strain to another.

Nippon Juigaku Zasshi, 1989 Oct, 51(5), 1003 - 10
Biological effects of lipopolysaccharide from Achromobacter stenohalis on lymphocytes and macrophages; Isogai H et al.; The immunopotentiating activities of lipopolysaccharide from Achromobacter stenohalis (A-LPS) were examined . A-LPS was structurally atypical and gave no endotoxin shock in A-LPS-inoculated mice . Analysis in vitro showed that A-LPS was a potent activator of both macrophages and B-lymphocytes . After macrophage stimulation with A-LPS, interleukin-1 (IL-1) secretion, interferon (IFN) production and chemiluminescence (CL) response were induced . A-LPS was a potent mitogen for spleen lymphocytes . However, induction of interleukin-2 (IL-2) secretion in T lymphocytes was not observed . These activities of A-LPS were similar to or higher than that of enterobacterial LPS.

J Biol Chem, 1989 Sep 25, 264(27), 16282 - 91
Cytoplasmic orientation and two-domain structure of the multidrug transporter, P-glycoprotein, demonstrated with sequence-specific antibodies; Yoshimura A et al.; The predicted cytoplasmic orientation and two-domain structure of the multidrug efflux pump P-glycoprotein were demonstrated with sequence-specific antibodies . We synthesized peptides corresponding to amino acid residues, Glu393-Lys408 (anti-P) and Leu1206-Thr1226 (anti-C) in P-glycoprotein from human mdr1 cDNA and used these peptides to produce polyclonal antibodies . From the primary structure of P-glycoprotein, and anti-C antibody is expected to recognize another position, Leu561-Thr581, in the duplicate structure of P-glycoprotein, but anti-P recognizes only one site . These antibodies bind to multidrug-resistant cells (KB-C2) with permeabilized plasma membrane but do not bind to nonpermeabilized KB-C2 cells or parental KB cells, supporting the predicted cytoplasmic orientation of these sequences . With immunoblotting of the membrane fractions from KB-C2 cells, a major 140-kDa polypeptide of the P-glycoprotein was detected with both anti-P and anti-C . Two minor polypeptides with molecular mass of 95 and 55 kDa were also detected . When membrane vesicles were digested mildly with trypsin, the amount of these two polypeptides increased . Anti-P detected only the 95-kDa polypeptide, and anti-C detected both 95- and 55-kDa polypeptides . Achromobacter lyticus protease I (lysyl endopeptidase) and Staphylococcus aureus V8 protease also produced two polypeptides with similar molecular weights . Absorption into lectin-agarose beads and labeling with {3H}glucosamine indicated that the 95-kDa polypeptide was glycosylated but that the 55-kDa polypeptide was not . These two polypeptides as well as P-glycoprotein were photoaffinity-labeled with a calcium channel blocker, {3H}azidopine, but most of the label was found in the 55-kDa polypeptide . The yield of labeled fragments from membrane vesicles photolabeled after digestion with trypsin was similar to that from membrane vesicles digested with trypsin after photolabeling . These data indicate 1) that the 95-kDa polypeptide is the fragment corresponding to the amino-terminal half of P-glycoprotein containing sugar chains; 2) that the 55-kDa polypeptide is the carboxyl-terminal half which was mainly labeled with {3H}azidopine; and 3) that P-glycoprotein has a relatively rigid structure with a small number of protease-sensitive sites and its global structure is not destroyed by tryptic cleavage.

J Clin Microbiol, 1989 Jul, 27(7), 1538 - 42
Application of gas-liquid chromatography to the routine identification of nonfermenting gram-negative bacteria in clinical specimens; Veys A et al.; A total of 430 strains of glucose-nonfermenting gram-negative bacteria representing 35 species were analyzed for their cellular fatty acid composition by gas-liquid chromatography (GLC) . On the basis of qualitative differences in their cellular fatty acid composition, these bacteria could be divided into 19 distinct chromatographic groups . Eight Pseudomonas species, Achromobacter xylosoxidans, group Vd, and Agrobacterium radiobacter were identified from their fatty acid compositions alone . The other glucose-nonfermenting gram-negative bacterial species studied here, classified within nine distinct GLC groups, were easily recognized by using the GLC fatty acid analysis supplemented with a limited number of conventional biochemical tests . The results support the hypothesis that bacterial fatty acid composition is rather specific and that qualitative GLC fatty acid analysis can be adapted in the clinical laboratory either to provide additional criteria for differentiation of closely related groups or to serve as a rapid and highly reproducible method for their routine identification.

J Bacteriol, 1989 Jul, 171(7), 4038 - 44
Cloning of a carbofuran hydrolase gene from Achromobacter sp . strain WM111 and its expression in gram-negative bacteria; Tomasek PH et al.; A 14-kilobase-pair (kbp) EcoRI DNA fragment that encodes an enzyme capable of rapid hydrolysis of N-methylcarbamate insecticides (carbofuran hydrolase) was cloned from carbofuran-degrading Achromobacter sp . strain WM111 . When used to probe Southern blots containing plasmid and total DNAs from WM111, this 14-kbp fragment hybridized strongly to a 14-kbp EcoRI fragment from the greater than 100-kbp plasmid harbored by this strain but weakly to EcoRI-digested total DNA from Achromobacter sp . strain WM111, indicating that the gene for N-methylcarbamate degradation (mcd) is plasmid encoded . Further subcloning localized the mcd gene on a 3-kbp ScaI-ClaI fragment . There was little or no expression of this gene in the alternative gram-negative hosts Pseudomonas putida, Alcaligenes eutrophus, Acinetobacter calcoaceticus, and Achromobacter pestifer . Western blotting (immunoblotting) of the protein products produced by low-level expression in P . putida confirmed that this 3-kbp fragment encodes the two 70+-kilodalton protein products seen in sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified carbofuran hydrolase.

J Biochem (Tokyo), 1989 Jun, 105(6), 986 - 91
Comparative studies on the primary structure of human cystatin as from epidermis, liver, spleen, and leukocytes; Takeda A et al.; We have studied the primary structure of human cystatin As from epidermis, liver, spleen, and leukocytes . These molecules were indistinguishable on PAGE in the presence and absence of SDS, by fast protein liquid chromatography (FPLC) chromatofocusing on a Mono P column, and in amino acid composition . The NH2- and COOH-terminal amino acid sequences of human cystatin As from epidermis, liver, and spleen were identical with those of human leukocyte cystatin A previously reported except for the lack of the NH2-terminal methionine residue in human epidermal cystatin A . The peptides obtained upon digestion of four human cystatin As with Achromobacter protease I (AP) showed identical peptide maps on HPLC except for different retention times of the NH2-terminal peptides . Furthermore, the amino acid compositions of corresponding separated peptide quartets were identical . We also determined the complete amino acid sequence of human epidermal cystatin A by sequencing peptides obtained from AP digestion and cyanogen bromide (CNBr) cleavage . It consisted of 97 amino acid residues, and was identical with those of human cystatin As from liver, spleen, and leukocytes except for the lack of the NH2-terminal methionine residue.

Rev Cubana Med Trop, 1989 May-Aug, 41(2), 274 - 83
{Identification of non-fermenting gram-negative bacteria . "Havana Center" Pediatric Teaching Hospital . December 1986 through May 1987}; Azahares Romero LE et al.; A study is made of non-fermenting gram-negative bacterial strains isolated at the Microbiology Lab, "Centro Habana" Pediatric Teaching Hospital . The strains studied come from various types of samples with prevalence of exudates and smears . A definite biotyping of 100% of strains is made . 90.0% are classified within the genus Pseudomonas . In addition, organisms of the genera Acinetobacter, Alcaligenes, Achromobacter, and Moraxella are identified . The most frequent species turned out to be Pseudomonas aeruginosa, which accounted for 68.4% of the total.

J Biochem (Tokyo), 1989 Apr, 105(4), 573 - 6
Identification of glutamic acid 186 affinity-labeled by 2,3-epoxypropyl alpha-D-glucopyranoside in soybean beta-amylase; Nitta Y et al.; Soybean beta-amylase was modified with 2,3-epoxypropyl alpha-D-{U-14C}glucopyranoside ({14C}alpha-EPG), a radioactive affinity-labeling reagent for beta-amylase, until it lost 95% of its enzyme activity . After S-carboxymethylation at pH 8.0 of SH groups, the modified enzyme was digested at pH 7.0 with Achromobacter protease I and the digest was fractionated by reverse-phase HPLC . A radioactive peptide was finally isolated and its amino acid sequence was determined to be 181Leu-Gly-Pro-Ala-Gly-Glu186 . Radioactivity derived from {14C}-alpha-EPG was found exclusively at Glu-186, the gamma-carboxyl group of which is esterified with the affinity label . It was concluded that the carboxylate of Glu-186 is a functional group at the catalytic site of soybean beta-amylase.

J Biol Chem, 1989 Mar 5, 264(7), 3832 - 9
The primary structure and structural characteristics of Achromobacter lyticus protease I, a lysine-specific serine protease; Tsunasawa S et al.; The complete amino acid sequence of Achromobacter lyticus protease I (EC 3.4.21.50), which specifically hydrolyzes lysyl peptide bonds, has been established . This has been achieved by sequence analysis of the reduced and S-carboxymethylated protease and of peptides obtained by enzymatic digestion with Achromobacter protease I itself and Staphylococcus aureus V8 protease and by chemical cleavage with cyanogen bromide . The protease consists of 268 residues with three disulfide bonds, which have been assigned to Cys6-Cys216, Cys12-Cys80, and Cys36-Cys58 . Comparison of the amino acid sequence of Achromobacter protease and other serine proteases of bacterial and mammalian origins has revealed that Achromobacter protease I is a mammalian-type serine protease of which the catalytic triad comprises His57, Asp113, and Ser194 . It has also been shown that the protease has 9- and 26-residue extensions of the peptide chain at the N and C termini, respectively, and overall sequence homology is as low as 20% with bovine trypsin . The presence of a disulfide bridge between the N-terminal extension Cys6 and Cys216 close to the putative active site in the C-terminal region is thought to be responsible for the generation of maximal proteolytic function in the pH range 8.5-10.7 and enhanced stability to denaturation.

Antibiot Khimioter, 1989 Jan, 34(1), 7 - 10
{Production of antibiotic substances by natural variants of the marine bacterium Vibrio Fischeri}; Baranova NA et al.; It was shown that under definite conditions there was competition between natural variants of sea bacteria belonging to V . fischeri . Natural variants of V . fischeri, strain 6 differed in their resistance to streptomycin and had different growth rates under conditions of limited aeration . Morphologically all the variants were identical . V . fischeri P-0, V . fischeri P-1 and V . fischeri P-2 were studied . The study revealed that V . fischeri P-0 produced a non-dialysing thermostable trypsin-sensitive substance inhibiting the growth of V . fischeri P-1 and V . fischeri P-2 . The maximum activity of the antibacterial substance was observed when V . fischeri P-0 was grown in a liquid medium with peptone and yeast extract without agitation at 26 degrees C . The inhibiting substance was also active against V . fischeri BKM B995 and V . fischeri P-7 isolated as a result of V . fischeri P-0 exposure to ethidium bromide . The substance had no effect on the following bacterial species: Aeromonas liquefaciens 301, Achromobacter liquefaciens, Pseudomonas putida 15, Pseudomonas fluorescence 7, Escherichia coli AH-32 and Staphylococcus aureus.

Bioorg Khim, 1989 Jan, 15(1), 5 - 17
{Catalytic component of calmodulin-independent adenylate cyclase from bovine brain . Isolation and determination of partial amino acid sequence}; Lipkin VM et al.; The catalytic component of calmodulin-independent adenylate cyclase of cattle cerebral cortex was solubilized and purified to the homogeneous state . The conditions for preparative obtaining of the enzyme on the column with immobilized antibodies to adenylate cyclase were found . These antibodies were proved to interact with the calmodulin-independent rather than the calmodulin-dependent form of the enzyme . Molecular mass of the calmodulin-independent adenylate cyclase determined electrophoretically is 140 +/- 10 kDa . Amino acid composition of the enzyme and sequences of its fragments (in total 300 amino acid residues) obtained upon treatment with lysyl-specific proteinase from Achromobacter liticus were determined . Clone containing a cDNA 605 bp insertion coding for the 183 amino acid residue fragment of adenylate cyclase was isolated from the bovine brain cDNA library . Homology of this fragment to the known sequences of Escherichia coli and Bordetella pertussis adenylate cyclases was revealed.

FEMS Microbiol Lett, 1989 Jan 1, 48(1), 61 - 4
Extracellular product of Nocardia amarae induces bacterial cell flocculation; Koizumi J et al.; The fact that Nocardia amarae YK1 produced a bacterial flocculation-inducing substance (designated as FIX) was discovered . FIX had a function of flocculating proliferous cells . FIX-induced flocculation was inhibited by making cells resting, but not completely by adding chloramphenicol . FIX worked widely on Gram-positive to -negative bacteria . In the presence of FIX, Achromobacter cycloclastus IAM1013, Acinetobacter calcoaceticus IAM1517, Bacillus subtilis IAM1069, Escherichia coli C600-1, E . coli IAM1239, Flavobacterium lutescens IAM1667, Klebsiella pneumoniae IAM1102, Micrococcus luteus IAM1313 and Pseudomonas putida IAM1002 formed flocs . B . cereus IAM1029, however, exhibited no flocculation.

G Batteriol Virol Immunol, 1989 Jan-Dec, 82(1-12), 34 - 8
{Bacterial infections of the ear: case contribution on clinical and microbiological aspects}; Cavallo GP et al.; Ciprofloxacin was given in oral doses of 250 mg . each 8 hours to 21 patients with otitis . The efficacy of treatment was assessed by clinical and bacteriological parameters . The 61.9% of isolated bacteria are opportunistic . A case of chronic otitis by Achromobacter xylosoxidans is described.

J Clin Microbiol, 1988 Nov, 26(11), 2425 - 6
Achromobacter xylosoxidans (Alcaligenes xylosoxidans subsp . xylosoxidans) meningitis associated with a gunshot wound; D'Amato RF et al.; The microbiological and clinical features of a case of Achromobacter xylosoxidans (Alcaligenes xylosoxidans subsp . xylosoxidans) meningitis associated with a gunshot wound are described . To our knowledge, this is the third confirmed case report of meningitis caused by this organism.

Appl Environ Microbiol, 1988 Nov, 54(11), 2874 - 6
Microbial transformation of the pyrethroid insecticides: permethrin, deltamethrin, fastac, fenvalerate, and fluvalinate; Maloney SE et al.; Pure cultures of Bacillus cereus, Pseudomonas fluorescens, and Achromobacter sp . were shown to transform five pyrethroid insecticides in the presence of Tween 80 . One of the major products, 3-phenoxybenzoic acid, was further transformed to 4-hydroxy-3-phenoxybenzoic acid . Permethrin was the most rapidly transformed of the pyrethroids, with a half-life of less than 5 days.

Microb Pathog, 1988 Nov, 5(5), 57 - 69
Identity of molecular structure of Shiga-like toxin I (VT1) from Escherichia coli O157:H7 with that of Shiga toxin; Takao T et al.; The primary structures of the A and B subunits of Shiga toxin and of Shiga-like toxin I (VT1), isolated from the culture supernatants of Shigella dysenteriae 1 and Escherichia coli O157:H7, respectively, were analyzed by Edman degradation of intact proteins and peptides in their digests with trypsin or Achromobacter protease I and also by fast atom bombardment mass spectrometry of the digests . The results indicated that the A and B subunits of Shiga toxin and Shiga-like toxin I have the same primary structures . The identity of their primary structures was confirmed by determining the nucleotide sequence of the gene encoding Shiga-like toxin I cloned from a Shiga-like toxin I converting phage . This nucleotide sequence was different from that reported by Jackson et al . (Microbial Pathogenesis 1987; 2: 147-153), by Calderwood et al . (Proc Natl Acad Sci USA 1987; 84: 4364-8) and by Grandis et al . (J Bacteriol 1987; 169: 4313-9) in one base at position 231, which was found to be adenine instead of thymine, which they reported . The amino acid residue at position 45 from the N-terminus of the A subunit of Shiga-like toxin I deduced from the nucleotide sequence determined in this study is threonine, which corresponds with that found by amino acid sequencing, whereas from previous reports by other investigators it is serine . Edman degradation of the intact A subunit of Shiga toxin indicated that the A subunit was nicked between Ala253 and Ser254 to form A1 and A2 fragments linked by a disulfide bond.

J Biol Chem, 1988 Oct 15, 263(29), 14625 - 8
Resonance Raman spectra of the copper-sulfur chromophores in Achromobacter cycloclastes nitrite reductase; Dooley DM et al.; Resonance Raman spectroscopy at ambient temperature and 77 K has been used to probe the structures of the copper sites in Achromobacter cycloclastes nitrite reductase . This enzyme contains three copper ions per protein molecule and has two principal electronic absorption bands with lambda max values of 458 and 585 nm . Comparisons between the resonance Raman spectra of nitrite reductase and blue copper proteins establish that both the 458 and 585 nm bands are associated with Cu(II)-S(Cys) chromophores . A histidine ligand probably is also present . Different sets of vibrational frequencies are observed with 457.9 nm (ambient) or 476.1 nm (77 K) excitation as compared with 590 nm (ambient) or 593 nm (77 K) excitation . Excitation profiles indicate that the 458 and 585 nm absorption bands are associated with separate {Cu(II)-S(Cys)N(His)} sites or with inequivalent and uncoupled cysteine ligands in the same site . The former possibility is considered to be more likely.

Eur J Biochem, 1988 Oct 1, 176(3), 683 - 97
Amino-acid sequence of ribonuclease T2 from Aspergillus oryzae; Kawata Y et al.; The amino acid sequence of ribonuclease T2 (RNase T2) from Aspergillus oryzae has been determined . This has been achieved by analyzing peptides obtained by digestions with Achromobacter lyticus protease I, Staphylococcus aureus V8 protease, and alpha-chymotrypsin of two large cyanogen bromide peptides derived from the reduced and S-carboxymethylated or S-aminoethylated protein . Digestion with A . lyticus protease I was successfully used to degrade the N-terminal half of the S-aminoethylated protein at cysteine residues . RNase T2 is a glycoprotein consisting of 239 amino acid residues with a relative molecular mass of 29,155 . The sugar content is 7.9% (by mass) . Three glycosylation sites were determined at Asns 15, 76 and 239 . Apparently RNase T2 has a very low degree of sequence similarity with RNase T1, but a considerable similarity is observed around the amino acid residues involved in substrate recognition and binding in RNase T1 . These similar residues may be important for the catalytic activity of RNase T2.

Anal Biochem, 1988 Oct, 174(1), 54 - 64
Generation of peptides suitable for sequence analysis by proteolytic cleavage in reversed-phase high-performance liquid chromatography solvents; Welinder KG; A methodological study of practical importance to protein sequencing has been carried out . Peptide mapping and sequence analysis of the cleavage products of reduced and carboxymethylated ribonuclease have been applied to the study of the activity and specificity of trypsin, chymotrypsin, elastase, lysyl endopeptidase (Achromobacter protease I), endoproteinase Arg-C (from mouse submaxillary gland), Staphylococcus aureus V8 protease, pepsin, and thermolysin in the presence of 20% methanol, ethanol, 2-propanol, and acetonitrile at 22 and 37 degrees C . The peptide bond specificities were retained, and the activities were generally unaffected or moderately reduced at 22 degrees C and pH 8 . At 37 degrees C the activity of chymotrypsin, endoproteinase Arg-C, V8 protease at pH 4, and pepsin was substantially reduced and decreased in the order methanol, ethanol, 2-propanol, and acetonitrile . The activity of thermolysin at 55 degrees C was reduced very little in the presence of 20% organic solvent and 50 mM Ca2+ . In low calcium and 20% 2-propanol at 22 degrees C the activity of thermolysin was restricted to the complete and specific cleavage of peptide bonds N-terminally of Phe, Ile, and Leu . The experiments suggest that secondary proteolytic digestions can be carried out directly in reversed-phase-HPLC fractions, and that organic cosolvents can be applied to control the degree of proteolysis . Moreover, the denaturing potential of these solvents might be useful in the degradation of proteins resistant to proteolysis, for example, in studies aimed at identification of disulfide bridges.

J Clin Microbiol, 1988 Sep, 26(9), 1901 - 3
Evaluation of 9-chloro-9-{4-(diethylamino)phenyl}- 9,10-dihydro-10-phenylacridine hydrochloride (C-390) in broth and agar media for identification of Pseudomonas aeruginosa; Fader RC et al.; Brain heart infusion broth and Mueller-Hinton agar containing the compound 9-chloro-9-{4-(diethylamino)phenyl}- 9,10-dihydro-10-phenylacridine hydrochloride (C-390) were evaluated as selective media for identifying Pseudomonas aeruginosa . Of the 192 Pseudomonas spp . and 68 additional oxidase-positive or glucose-nonfermenting gram-negative organisms tested, only P . aeruginosa (120 of 121 isolates) and certain strains of Pseudomonas cepacia, Aeromonas hydrophila, and Achromobacter xylosoxidans were able to grow in brain heart infusion broth containing 15 micrograms of C-390 per ml . When production of green or yellow-green pigment after overnight growth on Mueller-Hinton agar containing 30 micrograms of C-390 per ml was used as the criterion for identification, only P . aeruginosa produced a positive test (96% sensitivity and 100% specificity) . The agar medium, therefore, can provide a single differential medium for the overnight identification of P . aeruginosa.

Anal Biochem, 1988 Aug 1, 172(2), 420 - 6
A spectrophotometric assay for dissimilatory nitrite reductases; Hulse CL et al.; A spectrophotometric assay for dissimilatory nitrite reductases has been developed utilizing mammalian cytochrome c (equine heart) as reductant and spectrophotometric agent . The copper-containing nitrite reductase from Achromobacter cycloclastes has been shown to have apparent Km's for reduced cytochrome c and nitrite of 86 +/- 5 and 5.63 +/- 0.03 microM, respectively . The heme cd-containing enzyme from Pseudomonas stutzeri was shown to have apparent Km's for reduced cytochrome c and nitrite of 260 +/- 60 and 1.8 +/- 0.8 microM, respectively . This assay represents a simple, general method for consistently evaluating the activity of the copper- and heme cd-containing nitrite reductases that are capable of utilizing readily available mammalian cytochrome c as electron donor and should be useful for mechanistic studies of these enzymes.

J Hosp Infect, 1988 Jul, 12(1), 1 - 6
Outbreak of infection with Achromobacter xylosoxidans from contaminated intravascular pressure transducers; Gahrn-Hansen B et al.; Achromobacter xylosoxidans contaminating transducers caused 15 cases of hospital infection . In the eight patients with bacteraemia the interval from inoculation to fever was an average of 6.6 days . All the infected patients recovered . Computerization of laboratory records allowed retrieval of previous isolates, and review of clinical records focused the problem on patients with cardiac and aortic diseases . The problem arose from the re-use of disposable equipment after disinfection with a benzalcone.

Biochim Biophys Acta, 1988 Jun 29, 955(1), 43 - 9
New Achromobacter collagenase and its immunological relationship with a vertebrate collagenase; Nguyen TT et al.; Evidence is presented that Achromobacter iophagus produces two distinct collagenases . Achromobacter collagenases A and B were separated by high-performance liquid chromatography from partially purified enzyme . The main collagenase, A (EC 3.4.24.8), which has been already described, was eluted in the region of molecular mass 110-90 kDa . A minor collagenase B eluted in the region of 320 kDa, although in SDS-gel electrophoresis the apparent molecular masses of its main active forms were estimated as 55 and 110 kDa . The specificities of collagenases A and B are different . Collagenase A splits in its synthetic substrate Pz-Pro-Leu-Gly-Pro-DArg the bond Leu-Gly, collagenase B does not split this substrate . Both collagenases split bonds Gln-Gly and Leu-Gly in synthetic peptides DNP-Pro-Gln-Gly-Ile-Ala-Gly-Gln-DArg-OH and DNP-Pro-Leu-Gly-Ile-Ala-Gly-DArg-NH2, respectively . Collagenase B is twice as active as A on the native collagen type I . Both enzymes are inhibited by EDTA . The antibodies raised against the human tooth collagenase specifically inhibited the collagenase B, but did not influence the activity of collagenase A . These results indicate, to our knowledge for the first time, an immunological relationship between a bacterial and a vertebrate collagenase.

Eur J Clin Microbiol Infect Dis, 1988 Jun, 7(3), 428 - 31
Comparative in vitro activity of the new oral cephalosporin cefixime; Counts GW et al.; Cefixime was 8 to 10 times more active than cefaclor and augmentin against isolates of Escherichia coli, Klebsiella pneumoniae and Salmonella typhi, MIC90 values ranging from 0.06 to 0.25 micrograms/ml . However, none of these three drugs was particularly active against isolates of more resistant gram-negative bacilli such as Enterobacter, Serratia, Citrobacter, Acinetobacter, Providencia and Achromobacter spp . The lowest MIC values for gram-negative bacilli were seen with ciprofloxacin, except for isolates of Acinetobacter, where cotrimoxazole was the most active of the five drugs studied . Augmentin and ciprofloxacin exhibited the lowest MICs for isolates of streptococci and corynebacteria . Although cefixime may be among the most active oral beta-lactam drugs, it does not appear to be useful for treatment of infections caused by more resistant gram-negative bacilli.

Biochem Biophys Res Commun, 1988 May 31, 153(1), 74 - 80
Near stoichiometric, irreversible inactivation of bacterial collagenases by o-chloranil (3,4,5,6-tetrachloro-1,2-benzoquinone); Makinen PL et al.; The hydrogen-abstracting quinone derivative 3,4,5,6-tetrachloro-1,2-benzoquinone (o-chloranil) caused a strong, near stoichiometric, irreversible inactivation of the collagenases from Bacillus cereus, Clostridium histolyticum and Achromobacter iophagus . p-Chloranil was a weaker inactivator . o-Chloranil reacted rapidly with a site that affected substrate binding . Amino acid analyses of native and totally inactivated enzymes, and the pH-profile of inactivation suggest that the dissociated form of a tyrosine residue was modified.

J Biol Chem, 1988 May 15, 263(14), 6731 - 7
Characterization of the major protein-tyrosine-phosphatases of human placenta; Tonks NK et al.; In the preceding article (Tonks, N . K., Diltz, C . D., and Fischer, E . H . (1988) J . Biol . Chem . 263, 6722-6730), the purification of the major protein-tyrosine-phosphatases from human placenta, some to apparent homogeneity, was described . This report compares the characteristics of these enzymes and clearly identifies at least two distinct protein-tyrosine-phosphatase catalytic subunits . All were absolutely specific for phosphotyrosyl residues and showed no activity on any of the phosphoseryl/phosphothreonyl-containing proteins tested; they exhibited a high affinity for substrate with Km values in the submicromolar range . All were absolutely dependent on sulfhydryl compounds and appeared to contain at least one highly reactive cysteinyl residue essential for activity . Subtypes 1A and 1B could be distinguished by their response to polyanionic and polycationic compounds . The 1B enzymes were activated by EDTA, spermine, spermidine, and myelin basic protein to a greater extent than the 1A subtypes . Furthermore, they were inhibited by approximately 2 orders of magnitude lower concentrations of heparin (IC50 approximately 20 nM) and 1:1 or 4:1 poly (glutamate/tyrosine) (IC50 approximately 50 nM) than the 1A subtypes . Surprisingly, inhibition by the glutamate/tyrosine copolymers was strictly noncompetitive . Peptide mapping following digestion with Achromobacter protease I or Staphylococcus aureus V8 protease supported the view that, whereas protein-tyrosine-phosphatase subtypes 1A and 1B are different, their soluble and particulate counterparts are closely related structurally and are distinct from serine/threonine phosphatases 1 and 2A.

FEBS Lett, 1988 Apr 25, 231(2), 347 - 51
Primary structure of sheep prostaglandin endoperoxide synthase deduced from cDNA sequence; Yokoyama C et al.; The complete amino acid sequence of prostaglandin endoperoxide synthase from sheep vesicular gland has been deduced by cloning and sequence analysis of DNA complementary to its messenger RNA . The results were confirmed by digestion of the enzyme with carboxypeptidase Y and by automated Edman degradation of the intact enzyme polypeptide and peptide fragments obtained by limited proteolysis of the enzyme with Achromobacter proteinase I . Mature sheep prostaglandin endoperoxide synthase is shown to be composed of 576 amino acids with an Mr of 66,175 . The precursor peptide is predicted to contain a 24-residue signal peptide . The serine residue susceptible to acetylation by aspirin is found to be located near the C-terminus of the enzyme polypeptide.

J Biol Chem, 1988 Apr 25, 263(12), 5724 - 31
A phospholipase A2 in the supernatant fraction of rat spleen . Its similarity to rat pancreatic phospholipase A2; Tojo H et al.; Rat spleen supernatant contained two forms of calcium-dependent cellular phospholipase A2 which could be separated from each other by TEAE-cellulose chromatography . The phospholipase A2, named PLA2 S-1, present in the major flow-through fraction was purified to homogeneity . The structural and catalytic properties of splenic PLA2 S-1 were systematically compared with those of rat pancreatic phospholipase A2 . Structural evidence, including the sequence of the N-terminal 32 residues, peptide maps obtained on Achromobacter protease I digestion and cyanogen bromide cleavage, and the amino acid composition, showed the close similarity of the two enzymes . Their catalytic and immunochemical properties were also similar . These results demonstrated the existence of a pancreatic type phospholipase A2 in a non-pancreatic organ as a member of the cellular phospholipases A2 and suggest the potential functional involvement of pancreatic type phospholipase A2 in cellular phospholipid metabolism.

J Biochem (Tokyo), 1988 Apr, 103(4), 596 - 605
Amino acid sequence of a coagulant enzyme, flavoxobin, from Trimeresurus flavoviridis venom; Shieh TC et al.; The amino acid sequence of a coagulant enzyme, named flavoxobin, isolated from the venom of Trimeresurus flavoviridis (the habu snake) was determined by sequencing the S-pyridylethylated derivative of the protein and its peptides generated by chemical (cyanogen bromide and hydroxylamine) and enzymatic (clostripain, Staphylococcus aureus V8 protease, Achromobacter protease I, and elastase) cleavages . Hydrazinolysis was also employed to determine the C-terminal amino acid . The enzyme consisted of 236 amino acids and had a calculated molecular weight of 25,744 . Flavoxobin was found to be highly (69%) homologous in sequence to batroxobin, a coagulant enzyme from the venom of Bothrops atrox, and 27, 39, and 31% homologous to bovine thrombin, bovine trypsin, and human kallikrein, respectively . The sequence around the active site serine residue deduced from the homology relationship was Phe-Asp-Ser-Gly-Thr, which is different from the common sequence, Gly-Asp-Ser-Gly-Gly, for most serine proteases . Flavoxobin appears to be similar in secondary structure composition to batroxobin.

J Clin Microbiol, 1988 Mar, 26(3), 598 - 9
Achromobacter xylosoxidans (Alcaligenes xylosoxidans subsp . xylosoxidans) bacteremia associated with a well-water source: case report and review of the literature; Spear JB et al.; A case of community-acquired Achromobacter xylosoxidans bacteremia in a patient with metastatic breast carcinoma is described . The patient's home drinking water was identified as the source of her bacteremia . The case represents the first in which a community-acquired infection due to this organism has been attributed to a documented water source.

J Biol Chem, 1988 Feb 25, 263(6), 2651 - 7
Primary structure of chicken liver acetyl-CoA carboxylase deduced from cDNA sequence; Takai T et al.; The complete amino acid sequence of acetyl-CoA carboxylase from chicken liver has been deduced by cloning and sequence analysis of DNA complementary to its messenger RNA . The results were confirmed by Edman degradation of peptide fragments obtained by digestion of the enzyme polypeptide with Achromobacter proteinase I or staphylococcal serine proteinase . Chicken liver acetyl-CoA carboxylase is predicted to be composed of 2,324 amino acid residues, having a calculated molecular weight of 262,706 . The biotin carboxyl carrier protein domain is located in the middle region of the enzyme polypeptide . The amino-terminal portion of the acetyl-CoA carboxylase has been found to exhibit a homologous primary structure to that of carbamyl phosphate synthetase . Localization of possible functional domains including biotin carboxylase subsite in the acetyl-CoA carboxylase polypeptide is discussed.

APMIS, 1988 Feb, 96(2), 185 - 7
Studies on ampicillin resistance in Achromobacter xylosoxidans . Brief report; Gahrn-Hansen B et al.; Population analyses of susceptibility to ampicillin in ampicillin-susceptible and ampicillin-resistant strains of Achromobacter xylosoxidans revealed the existence of ampicillin-resistant subpopulations in ampicillin-susceptible isolates . Bacteria resistant to a concentration four times the one that inhibited the majority population had a frequency of 10(-3) to 10(-4) . Strains isolated from aqueous environments are often found susceptible to ampicillin, while sporadic cases of infections with A . xylosoxidans are often caused by ampicillin-resistant strains . We suggest that the isolation from clinical specimens of ampicillin-susceptible strains, therefore, may be an indication of nosocomial infections due to recently contaminated aqueous solutions or medical equipment.

Infect Control Hosp Epidemiol, 1988 Feb, 9(2), 84 - 7
Nosocomial Achromobacter xylosoxidans infections; Schoch PE et al.; A xylosoxidans is being recognized as an important nosocomial pathogen . As more patients are rendered immunosuppressed by chemotherapy, this organism's increasing role in hospital-acquired infections will be assured . Achromobacter is a water-borne organism, highly resistant to most antibiotics, and even to some disinfectant solutions, and easily establishes itself in the hospital aquatic environment . Achromobacter infections and outbreaks should be recognized and approached as serious problems requiring the institution of appropriate infection control measures . A xylosoxidans infections should be empirically treated with a combination of a third generation cephalosporin and TMP-SMX pending susceptibility testing.

J Biol Chem, 1988 Jan 15, 263(2), 839 - 49
Complete amino acid sequence of rat liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase; Lively MO et al.; The complete amino acid sequence of 6-phospho-fructo-2-kinase/fructose-2,6-bisphosphatase from rat liver was determined by direct analysis of the S-carboxamidomethyl protein . A complete set of nonoverlapping peptides was produced by cleavage with a combination of cyanogen bromide and specific proteolytic enzymes . The active enzyme is a dimer of two identical polypeptide chains composed of 470 amino acids each . The NH2-terminal amino acid residue of the polypeptide chain was shown to be N-acetylserine by fast atom bombardment mass spectrometry of the purified N-terminal tetradecapeptide isolated after cleavage of the intact S-carboxamidomethylated protein with lysyl endoproteinase (Achromobacter protease I) . Alignment of the set of unique peptides was accomplished by the analysis of selected overlapping peptides generated by proteolytic cleavage of the intact protein and the larger purified cyanogen bromide peptides with trypsin, Staphylococcus aureus V8 protease, and lysyl endoproteinase . Four nonoverlapping peptides were aligned by comparison with the amino acid sequence predicted from a partial cDNA clone encoding amino acid positions 166-470 of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (Colosia, A.D., Lively, M., El-Maghrabi, M . R., and Pilkis, S . J . (1987) Biochem . Biophys . Res . Commun . 143, 1092-1098) . The nucleotide sequence of the cDNA corroborated the peptide sequence determined by direct methods . A search of the Protein Identification Resource protein sequence database revealed that the overall amino acid sequence appears to be unique since no obviously homologous sequences were identified . However, a 100-residue segment of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (residues 250-349), including the active site histidine residue of the bisphosphatase domain, was found to be homologous to the active site regions of yeast phosphoglycerate mutase and human bisphosphoglycerate mutase.

Connect Tissue Res, 1988, 17(2), 153 - 8
Bacterial collagenase and collagen identification; Berry L et al.; A method is described for the cleavage of collagenous molecules by bacterial collagenase in the presence of sodium dodecyl sulphate (SDS) and urea . Comparison of three commercially available preparations of bacterial collagenase showed that the most efficient cleavage under these conditions was by the enzyme isolated from Achromobacter iophagus (E.C . 3.4.24.8) . No non-specific proteinase activity was apparent in conditions where all collagen types showed some susceptibility to attack . This method represents a simple one-stage identification of collagenous molecules in complex mixtures of proteins and where limited amounts of a protein are available.

J Clin Microbiol, 1987 Oct, 25(10), 1952 - 5
Diversity of plasmids in Achromobacter xylosoxidans isolates responsible for a seemingly common-source nosocomial outbreak; Arroyo JC et al.; Achromobacter xylosoxidans, an uncommon yet highly resistant opportunistic pathogen, was isolated from nine hospitalized patients during an 8-month period . It had been isolated from only seven patients with either nonfatal infection or colonization from 1981 to 1984 . From June 1985 to January 1986, A . xylosoxidans was isolated 18 times from seven different sites (sputum, 7 times; urine, 4 times; blood, 3 times; and lung, pleural fluid, wound tissue, and tracheal aspirate, 1 time each) . Four patients died, including the three with bacteremia . All but two patients had nosocomial infections and either were on the same ward or were cared for by the same staff members . Eleven A . xylosoxidans strains yielded eight distinct plasmids (8, 21, 23, 26, 38, 50, 51, and 64 megadaltons) . Whole-cell peptide patterns of 10 of these strains were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Isolates from the same patient contained the same plasmids and had identical peptide patterns but differed from other strains in both parameters . Plasmids were absent from the two community-acquired isolates . Although nosocomial strains showed similar antibiotic resistance patterns (only moxalactam and ticarcillin-clavulanic acid were uniformly active) and cross-contamination was strongly suggested epidemiologically, results of plasmid and peptide analyses did not support the possibility of a single-strain outbreak.

Rev Infect Dis, 1987 Sep-Oct, 9(5), 1001 - 5
Achromobacter xylosoxidans bacteremia; Mandell WF et al.; Achromobacter xylosoxidans is a rare cause of bacteremia . A case of community-acquired pneumonia and bacteremia due to Achromobacter in a patient with concomitant pulmonary tuberculosis is reported herein . The majority of patients who have developed achromobacter bacteremia have had a predisposition to infection (although the predisposing conditions have been diverse) . Immunosuppression has been reported in only one of the seven patients with achromobacter bacteremia for whom detailed information is available . Achromobacter is usually resistant to ampicillin, cephalosporins, and aminoglycosides . Antipseudomonal penicillins and trimethoprim-sulfamethoxazole inhibit most isolates . Multiple-drug resistance is common, and optimal therapy is not known.

Proc Natl Acad Sci U S A, 1987 Aug, 84(16), 5610 - 4
Amino acid sequence of the von Willebrand factor-binding domain of platelet membrane glycoprotein Ib; Titani K et al.; We report the amino acid sequence of a 299-residue segment from the alpha chain of the human platelet membrane glycoprotein Ib . This includes the complete sequence of the amino-terminal tryptic fragment of 290 residues comprising the von Willebrand factor-binding domain . Two primary sets of overlapping fragments were obtained by cleavage of the S-carboxymethylated protein at methionyl and lysyl bonds following treatment with cyanogen bromide and Achromobacter protease I, respectively . Additional fragments were obtained by treatment of native glycocalicin with trypsin, Staphylococcus aureus V8 protease, and Serratia marcescens protease . Analysis of all these fragments provided data that allowed determination of the continuous sequence corresponding to approximately half of the alpha-chain polypeptide . This region of glycoprotein Ib is largely hydrophobic and contains only two N-linked and one O-linked carbohydrate chains . A hydrophilic region exists between residues 215 and 299, which contains a cluster of 10 negatively charged residues at 269-287 . This area is likely to attract positively charged molecules . The hydrophilic, highly glycosylated (at serine and threonine residues) region corresponding to the previously described "macroglycopeptide" and representing the carboxyl-terminal half of the alpha chain is likely to begin at residue 292 . The determined sequence of the alpha chain of glycoprotein Ib contains a region (residues 29-193) with seven repeats, which is indicative of gene duplication and is highly homologous to human leucine-rich alpha 2-glycoprotein . This protein sequence agrees completely with that deduced from the cDNA sequence reported by Lopez et al . {Lopez, J.A., Chung, D.W., Fujikawa, K., Hagen, F.S., Papayannopoulou, T . & Roth, G.J . (1987) Proc . Natl . Acad . Sci . USA 84, 5615-5619}.

J Biochem (Tokyo), 1987 Jul, 102(1), 111 - 22
Bacterial synthesis of recombinant alpha-human atrial natriuretic polypeptide; Saito Y et al.; The high-level synthesis of alpha-human atrial natriuretic polypeptide hormone in Escherichia coli has been achieved based on the idea that the yield of a small, basic and unstable polypeptide, such as the natriuretic polypeptide, would be improved by fusion with an appropriate protective polypeptide to construct a neutral fused polypeptide . We prepared an expression vector, pCLaHtrp3t, coding a neutral polypeptide containing 130 amino acid residues in which the polypeptide hormone was fused to a newly designed protective polypeptide through lysine as an enzymatically cleavable residue . The fused polypeptide was synthesized at the high level of 32% of total cellular proteins and at 4.7 X 10(6) molecules per single cell . It was recovered as cellular insoluble fraction and purified to homogeneity . For the isolation of the peptide hormone from the resultant fused polypeptide, Achromobacter protease I, a lysine-specific endopeptidase was used, because it has sufficient activity even in 8 M urea . The recombinant natriuretic polypeptide was indistinguishable from native alpha-human atrial natriuretic polypeptide as regards amino acid sequence as well as biological activity.

Am J Ophthalmol, 1987 Jun 15, 103(6), 745 - 8
Chronic bacterial endophthalmitis; Ficker L et al.; We studied a specific syndrome of uveitis secondary to intraocular bacterial pathogens of low virulence after extracapsular cataract extraction and intraocular lens implantation in three eyes . The onset of photophobia, visual impairment, conjunctival redness, and uveitis was delayed for four days to 12 weeks after surgery . Chronic inflammation persisted for five weeks to 16 months before a definitive diagnosis was made . Signs and symptoms were suppressed by administration of topical and systemic corticosteroids . Intraocular biopsy and antibiotic injection both established the cause as bacterial endophthalmitis and resulted in resolution of signs and symptoms . Staphylococcus epidermidis was cultured in two eyes and Achromobacter was cultured in one.

Pathol Biol (Paris), 1987 Jun, 35(5 Pt 2), 839 - 42
{Treatment of post-traumatic and post-neurosurgical bacterial meningitis with ceftriaxone alone or in combination with fosfomycin}; May T et al.; From 1984 to 1986, 13 patients (10 adults, 3 children) with bacterial meningitis following neurosurgery or traumatism were given ceftriaxone alone 6 times at a dose of 40 mg/kg one IV injection per day, or in association 7 times with fosfomycin at a dose of 200 mg/kg/day, 3 IV perfusions every 4 h . The bacteriological diagnosis was confirmed in 9 cases (3 Staphylococcus aureus, 4 Streptococcus pneumoniae, 1 Klebsiella, 1 Peptococcus) . In vitro neither synergy nor antagonism were observed between the two antimicrobial agents . The acute infections episode resolved in all patients except on who died with a negative CSF culture . One superinfection meningitis with Achromobacter was seen . CSF concentrations of ceftriaxone were assayed and found to be comparable with those reported by most authors . Tolerance was excellent for all our patients.

Biochem Biophys Res Commun, 1987 May 29, 145(1), 563 - 8
Kinetic studies of the copper nitrite reductase from Achromobacter cycloclastes and its interaction with a blue copper protein; Kashem MA et al.; Transient state, burst and steady state kinetics of reactions of the blue copper nitrite reductase (NIR) and blue copper protein from Achromobacter cycloclastes are investigated . The two copper-containing species are reacted with each other and where possible with dithionite, ascorbate and nitrite . Both copper proteins are fully reduced by dithionite with both S2O4(2-) and SO2- . species active . NIR is only partially reduced by ascorbate in an unusual biphasic reaction consistent with complete reduction of type-one copper followed by partial reduction of type-two copper . The rate of reduction of the type-one copper is accelerated using phenazine methosulfate as mediator . Nitrite can oxidize dithionite-reduced NIR but cannot reduce oxidized NIR . Rate constants were determined for all observed reactions.

Biochemistry, 1987 Apr 21, 26(8), 2189 - 94
Amino acid sequence of sialic acid binding lectin from frog (Rana catesbeiana) eggs; Titani K et al.; The complete amino acid sequence of sialic acid binding lectin from frog (Rana catesbeiana) egg is presented . The 111-residue sequence was determined by the analysis of peptides generated by digestion of the S-carboxymethylated protein with Achromobacter protease I, chymotrypsin, or cyanogen bromide . The sequence is unique and not homologous to any known protein sequence . The protein may represent a new type of lectin.

J Biol Response Mod, 1987 Apr, 6(2), 194 - 204
Resistance to biological and chemical challenge in rodents treated with xerosin II, a natural product of Achromobacter xerosis; Boylan ES et al.; The biological response-modifying activity of acid-precipitable material from Achromobacter xerosis was first described as suppression of viral pneumonia in mice . Later, this acid-precipitable material (xerosin) was found to have antiinflammatory activity and to induce tumor regression in chickens infected with Rous sarcoma virus . Here, we report further purification of xerosin resulting in a product (xerosin II) that retains high biological activity against viral and endotoxin-induced pneumonia in mice . In addition, we describe new activities of xerosin II in two rat tumor systems . Female CD rats received gastric intubations of 7,12-dimethylbenz(a)anthracene; 2 weeks later, half began 4 weeks of treatment with xerosin II, while others received saline only . Xerosin II treatment significantly delayed the appearance of the first palpable mammary tumors per rat . In female F344 rats implanted with the 13762 mammary tumor, 4 weeks of xerosin II treatment prolonged the survival of rats by an average of 5-11 days (12-24%) in two separate trials . Tumor growth and incidence of metastasis appeared unaffected by xerosin II treatment . Thus, this refined bacterial extract proved to be a potent biological response modifier in four different rodent systems.

Blood, 1987 Feb, 69(2), 560 - 4
Purification and partial amino acid sequence of human platelet membrane glycoproteins IIb and IIIa; Hiraiwa A et al.; The glycoprotein (GP) IIb-IIIa complex was isolated from human platelet membranes by immunoaffinity chromatography using a monoclonal antibody specific for GP IIb-IIIa . GP IIb and IIIa were further separated in the presence of sodium dodecyl sulfate (SDS) by gel filtration high-performance liquid chromatography (HPLC) . Two cycles of this procedure yielded almost complete separation of homogeneous preparations of GP IIb and IIIa . Each protein was then digested with lysyl endopeptidase (Achromobacter protease I), which cleaves at the carboxyl side of lysine residues, and the resulting oligopeptides from GP IIb and IIIa were fractionated with HPLC using a C18 reverse-phase column . Comparison of the elution profiles showed no obvious homology between the two proteins . Amino acid sequences of selected oligopeptides from each glycoprotein were determined using a gas-phase protein sequencer . Sixty amino acid residues (26 residues for IIb and 34 residues for IIIa) were identified.

Chemioterapia, 1987 Feb, 6(1), 32 - 7
Emerging microorganisms in cystic fibrosis; Fabbri A et al.; Pseudomonas aeruginosa is the most common bacterial isolate obtained from patients with cystic fibrosis of the lungs . Recently, however, new multiresistant organisms have emerged, whose identification may be difficult and whose pathogenic role proves hard to define . Of the 71 strains isolated from 24 patients with cystic fibrosis during acute flareups of pulmonary symptoms, 48 turned out to be Pseudomonas aeruginosa (67.6%); 11 were Pseudomonas non-aeruginosa (15.5%); and 12 were Achromobacter xylosoxidans (16.9%) . Each bacterial isolate was tested for sensitivity to nine antibiotics (ceftazidime, azlocillin, piperacillin, aztreonam, cefsulodin, cefoperazone, amikacin, tobramycin, and sisomycin) in terms of minimum inhibitory concentration and minimum bactericidal concentration values . In this series, Achromobacter xylosoxidans proved the species least responsive to treatment, and ceftazidime the most active antibiotic both against Achromobacter and against strains of the genus Pseudomonas . Twenty-three different associations of ceftazidime with aminoglycosides, tested for activity on the multiresistant strains, failed to show synergism of action.

J Biochem (Tokyo), 1987 Jan, 101(1), 111 - 21
Amino acid sequence of thermostable direct hemolysin produced by Vibrio parahaemolyticus; Tsunasawa S et al.; The complete amino acid sequence of the subunit of thermostable direct hemolysin, a dimeric protein composed of identical subunits isolated from Vibrio parahaemolyticus, was determined by sequencing BrCN-peptides, their tryptic peptides, and overlaps obtained by Achromobacter protease I digestion . The subunit consists of 165 amino acid residues with the sole disulfide bond between Cys 151 and Cys 161 . It is deduced that the biologically active hemolysin is formed by noncovalent association of subunits which are not linked together by disulfide bonds . The primary structure of hemolysin elucidated in the present study is essentially the same as that deduced from the nucleotide sequence of a gene encoding the protein but differs in 9 amino acid residues, suggesting the possibility of the presence of multiple genes for the thermostable direct hemolysin in Vibrio parahaemolyticus.

Biochem J, 1986 Dec 15, 240(3), 803 - 10
Influence of temperature on the enzymic semisynthesis of human insulin by coupling and transpeptidation methods; Morihara K et al.; The influence of temperature of enzymic semisynthesis of human insulin ester was determined by using coupling and transpeptidation methods with trypsin and Achromobacter lyticus proteinase I as catalysts . The optimal reaction conditions were studied at the selected temperatures of 25, 12 and 4 degrees C . The results showed that the synthesis rates by both methods with trypsin increased as the temperature increased, but the final product yield correspondingly decreased . Therefore the reaction with trypsin should be done below 12 degrees C, preferably at 4 degrees C . This agrees well with the stability of trypsin at these temperatures . When the catalyst was Achromobacter lyticus proteinase I, no such complex temperature effects were observed, and the findings indicated that the reactions should be conducted below 37 degrees C for enzyme stability.

Infection, 1986 Nov-Dec, 14(6), 279 - 82
Infections due to Achromobacter xylosoxidans . Case report and review of the literature; Chandrasekar PH et al.; Achromobacter xylosoxidans is an uncommon nosocomial pathogen known to cause many serious infections . A 69-year-old woman with diabetes mellitus and chronic renal failure was admitted with pulmonary edema . The patient developed fever and pulmonary infiltrate with bilateral pleural effusions while she was on a respirator in the intensive care unit . Culture of sputum, pleural fluid and blood grew A . xylosoxidans . Bilateral chest tubes were inserted and the patient was treated for one month with piperacillin and trimethoprim-sulfamethoxazole . Gradual response, both clinically and radiologically, was noted after prolonged therapy . A review of the literature on infections due to A . xylosoxidans, the unique susceptibility pattern of the organism to various antibiotics and the use of combination therapy in Achromobacter infections are discussed.

Eur J Biochem, 1986 Oct 15, 160(2), 413 - 8
New thiol inhibitor of Achromobacter iophagus collagenase . Specificity of the enzyme's S3' subsite; Yiotakis A et al.; New synthetic mercaptotripeptides (HS-CH2-CH2-CO-Pro-Yaa) which inhibit Achromobacter iophagus collagenase were produced in order to obtain more powerful bacterial collagenase inhibitors than currently available, and to investigate the specificity of the S3' subsite of the enzyme . Since similar binding constants were found for inhibitors carrying uncharged residues of various sizes in the P3' position (Yaa = Ala, Leu, Phe, Pro, Hyp) steric hindrance at the collagenase S3' appears relatively limited . The compound (HS-CH2-CH2-CO-Pro-Arg), which carries an arginine residue in the position P3' and had the highest inhibition constant of the series tested (Ki = 0.5 microM), proved to be the strongest inhibitor so far reported in the literature . The weakest in the present series was the compound (HS-CH2-CH2-CO-Pro-Asp) which carries an aspartic residue in position P3' and had a Ki = 70 microM . The present work revealed that the charged groups in the P3' position play a key role in the interaction of the inhibitors with the enzyme.

Int J Pept Protein Res, 1986 Oct, 28(4), 411 - 9
Enzymatic semisynthesis of {LeuB30} insulin; Sakina K et al.; Experimental conditions for the preparation of {LeuB30} insulin by coupling of des-AlaB30 insulin with Leu-OBu(t) were determined using Achromobacter protease I and trypsin as catalysts . Successful coupling required a large excess of the amine component (0.8 M), a high concentration of organic cosolvent (35-50%) and neutral pH of the reaction mixture . The coupling yield of Achromobacter protease I after 24 h at 37 degrees C was almost the same or a little higher than that at 25 degrees C . With trypsin, the coupling yield at 37 degrees C after 24 h was considerably lower than at 25 degrees C . This was partly ascribed to the difference in concentration of organic cosolvent at 37 degrees C and 25 degrees C; 35% and 50%, respectively, or possibly of enzyme stability at these temperatures . The maximum product yield was about 90% with both enzymes under optimal conditions . A preparative scale experiment was performed with Achromobacter protease I; the yield of {LeuB30} insulin was 51% using porcine insulin as the starting material . This semisynthetic insulin was identified by HPLC and amino acid analysis . No difference was observed in CD spectra between {LeuB30} insulin and human insulin.

J Biochem (Tokyo), 1986 Jun, 99(6), 1749 - 52
Purification and some properties of cytochrome c' from a strain of Achromobacter xylosoxidans; Shidara S et al.; Cytochrome c' was crystallized from Achromobacter xylosoxidans GIFU 543 . The cytochrome was a basic protein and its molecular weight was 28,000 . The pyridine ferrohemochrome showed absorption peaks at 415, 521, and 551 nm . The absorption spectra of the oxidized and reduced forms at neutral pH were almost the same as those of other cytochromes c' reported already . The reduced cytochrome c' reacted with CO and NO, and the NO complex showed a characteristic absorption spectrum . The midpoint redox potential of the hemoprotein was measured to be + 110 mV at pH 7.2.

J Biochem (Tokyo), 1986 May, 99(5), 1289 - 98
Amino acid sequence of copper,zinc-superoxide dismutase from spinach leaves; Kitagawa Y et al.; The complete amino acid sequence of Cu,Zn-superoxide dismutase (SOD) from spinach leaves has been determined on the basis of peptides obtained by cyanogen bromide (BrCN) cleavage and by enzymic hydrolyses with Achromobacter lyticus lysylendopeptidase, Staphylococcus aureus V8 protease, trypsin, and thermolysin . The spinach SOD consists of a total of 154 amino acid residues with alanine as the amino(N)-terminus and valine as the carboxy(C-)terminus . The present sequence, which has been established for the enzyme from a plant, is also highly homologous to those of the enzymes from other species . Especially, the residues essential for metal binding and enzyme activity have been extensively conserved among all of the Cu,Zn-SODs hitherto analyzed.

Biochem Cell Biol, 1986 May, 64(5), 394 - 9
Electron paramagnetic resonance studies of heme c and its nitrosyl derivative in Vibrio (Achromobacter) fischeri nitrite reductase; Sadana JC et al.; Interactions of Vibrio (formerly Achromobacter) fischeri nitrite reductase were studied by electron paramagnetic resonance spectroscopy . The spectrum of the oxidized enzyme showed a number of features which were attributed to two low-spin ferric hemes . These comprised an unusual derivative peak at g = 3.7 and a spectrum at g = 2.88, 2.26, and 1.51 . Neither heme was reactive in the oxidized state with the substrate nitrite and with cyanide and azide . When frozen under turnover conditions (i.e., reduction in the presence of excess nitrite), the enzyme showed the spectrum of a nitrosyl heme derivative . The g = 2.88, 2.26, and 1.51 signals reappeared partially on reoxidation by nitrite, indicating that the nitrosyl species which remained arose from the g = 3.7 heme . The nitrosyl derivative showed a 14N nuclear hyperfine splitting, Az = 1.65 mT . The nitrosyl derivative was produced by treatment of the oxidized nitrite reductase with nitric oxide or hydroxylamine . Exchange of nitric oxide between the nitrosyl derivative and NO gas in solution was observed by using the {15N}nitrosyl compound . A possible reaction cycle for the enzyme is discussed, which involves reduction of the enzyme followed by binding of nitrite to one heme and formation of the nitrosyl intermediate.

J Biochem (Tokyo), 1986 Feb, 99(2), 407 - 22
Complete amino acid sequence of NADH-cytochrome b5 reductase purified from human erythrocytes; Yubisui T et al.; The complete amino acid sequence of soluble NADH-cytochrome b5 reductase purified from human erythrocytes was determined . The enzyme, which contained 8 methionine residues, was cleaved by cyanogen bromide . The resulting nine peptides were separated by gel filtration and purified further by high-performance liquid chromatography . The purified peptides were sequenced by automated Edman degradation . Three large CNBr peptides, residues 1-101, 109-151, and 169-231, were further fragmented with trypsin, Staphylococcus aureus V8 protease or a lysyl endopeptidase of Achromobacter lyticus . The peptides obtained from the tryptic digest of citraconylated FAD-depleted apoprotein completed the alignments of the other peptides . The enzyme was composed of 275 amino acid residues . The 4 functionally important cysteine residues were located in the COOH-terminal portion . The molecular weight of the protein was calculated to be 31,260 without FAD . A prediction of the secondary structure was made by the method of Chou and Fasman . The protein was hydrophilic as a whole (43% polarity), but some regions were rich in hydrophobic residues . From the sequence homology of this enzyme with the pyridine nucleotide-binding sites of other flavoproteins, three candidates for the FAD and NADH-binding domains were suggested.

J Biochem (Tokyo), 1986 Jan, 99(1), 281 - 9
Amino acid sequence of Trimeresurus flavoviridis phospholipase A2; Tanaka S et al.; The amino acid sequence of phospholipase A2 from the venom of Trimeresurus flavoviridis (the Habu snake) was determined . The enzyme subunit has a molecular weight of 13,764 and consists of a single polypeptide chain of 122 amino acids and seven disulfide bonds . The fragmentation was conducted by digesting the reduced and S-carboxymethylated derivative of the protein with Achromobacter protease I, chymotrypsin, and trypsin, respectively . Achromobacter protease I peptides were used for alignment and to establish overlaps over chymotryptic and tryptic peptides . The automated Edman degradation of the S-carboxymethylated protein, which was extended to the N-terminal 30 amino acid residues, supplemented the deletions found with the enzymatic peptides alone . T . flavoviridis phospholipase A2 was found to be highly (65-67%) homologous in sequence to the enzymes from T . okinavensis, Crotalus adamanteus, and Crotalus atrox (viperid family) and less (35-44%) homologous to those from elapid snakes and mammalian pancreas . The T . flavoviridis enzyme appears to be similar in secondary structure composition to the C . atrox enzyme.

Ann Biol Clin (Paris), 1986, 44(2), 176 - 80
{Peptide inhibitors of a bacterial collagenase}; Dive V et al.; The properties of the cleavage site of collagen by collagenases remain discussed . The authors report their studies on the active site of the collagenase of a bacteria, achromobacter, based on two types of methods . The first type uses synthetic substrates, the second one enzymatic inhibitors . The results of both methods are reviewed . The inhibitors seem more sensitive than substrates to study the relation between structure and activity of the enzyme.

J Clin Microbiol, 1985 Sep, 22(3), 470 - 1
Neonatal meningitis caused by Achromobacter xylosoxidans; Namnyak SS et al.; The clinical and bacteriological findings in a case of neonatal meningitis caused by Achromobacter xylosoxidans are presented . This appears to be only the second report of meningitis caused by this species.

J Biochem (Tokyo), 1985 Sep, 98(3), 585 - 603
Subtilosin A, a new antibiotic peptide produced by Bacillus subtilis 168: isolation, structural analysis, and biogenesis; Babasaki K et al.; Subtilosin A, a new antibiotic produced by Bacillus subtilis 168, was extracted from culture medium with n-butanol and purified to homogeneity by a combination of gel filtration and thin-layer chromatography . The yield was 5.5 mg from a liter of culture . It had bacteriocidal activity against some gram-positive bacteria . Amino acid analysis and mass spectrometry showed that it was a peptide with a molecular weight of 3398.9, consisting of 32 usual amino acid and some non-amino acid residues . Its amino- and carboxyl-termini were blocked . By analysis of the fragments obtained by partial acid hydrolysis, as well as by chymotryptic and thermolysin digestions of reduced and S-carboxymethylated samples and Achromobacter protease I digestion of performic acid-oxidized samples, the amino acid sequence was determined to be as follows: X-Gly-Leu-Gly-Leu-Trp-Gly-Asn-Lys-Gly-Cys-Ala-Thr-Cys-Ser-(sequence; see text) Ile-Gly-Ala-Ala-Cys-Leu-Val-Asp-Gly-Pro-Ile-Pro-Asp-Glx-Ile-Ala-Gly-Ala . The analyses of cross-linking structures revealed that there were linkages between the amino- and carboxyl-termini and between the Cys-19 and the Glx-28 residues through an unknown residue with a residue weight of 163 . Consequently, subtilosin A was deduced to be a cyclic peptide antibiotic with a novel cross-linking structure . The production of subtilosin A begins at the end of vegetative growth and finishes before spore formation . Studies on the correlation between the production of subtilosin A and spore formation with decoyinine in the original strain and in asporogenous mutants of B . subtilis 168 suggested that there was no close correlation between the two phenomena . The production of subtilosin A was repressed by inhibitors of protein and RNA synthesis in contrast to that of many other antibiotic peptides, suggesting that it is synthesized by the mechanism of usual protein synthesis.

J Biochem (Tokyo), 1985 Sep, 98(3), 799 - 805
Exposed tyrosine residues of lambda cro repressor protein evidenced by nitration and photo CIDNP experiments; Shirakawa M et al.; The tyrosine residues of lambda cro repressor were partially nitrated with tetranitromethane under mild conditions . After digestion by Achromobacter protease I, the extent of nitration was determined by HPLC and amino acid analysis . Tyr 26 was most easily nitrated and Tyr 51 followed it . Tyr 10 was resistant to nitration . By comparison of the proton magnetic resonance spectrum of the partially nitrated cro protein with the above result, the aromatic proton resonances of the tyrosine side chains could be assigned to individual tyrosine residues . The extent of nitration is parallel to the accessibility to a flavin dye as measured by photo CIDNP (chemically induced dynamic nuclear polarization).

FEBS Lett, 1985 Jul 1, 186(1), 41 - 5
Complete amino acid sequence of bovine colostrum low-Mr cysteine proteinase inhibitor; Hirado M et al.; The complete amino acid sequence of bovine colostrum cysteine proteinase inhibitor was determined by sequencing native inhibitor and peptides obtained by cyanogen bromide degradation, Achromobacter lysylendopeptidase digestion and partial acid hydrolysis of reduced and S-carboxymethylated protein . Achromobacter peptidase digestion was successfully used to isolate two disulfide-containing peptides . The inhibitor consists of 112 amino acids with an Mr of 12787 . Two disulfide bonds were established between Cys 66 and Cys 77 and between Cys 90 and Cys 110 . A high degree of homology in the sequence was found between the colostrum inhibitor and human gamma-trace, human salivary acidic protein and chicken egg-white cystatin.

Diagn Microbiol Infect Dis, 1985 Mar, 3(2), 149 - 58
Occurrence and antimicrobial susceptibility of gram-negative nonfermentative bacilli in cystic fibrosis patients; Klinger JD et al.; Isolation of nonfermentative gram-negative bacilli (other than Pseudomonas aeruginosa) from respiratory tract cultures of cystic fibrosis (CF) patients has increased in recent years . Species recovered include Pseudomonas cepacia, P . maltophilia, P . fluorescens/putida, P . alcaligenes, P . pseudoalcaligenes, P . stutzeri, Acinetobacter spp., Achromobacter xylosoxidans, Flavobacterium spp., and CDC groups IVe and Ve . Although colonization with most of these organisms is sporadic, P . cepacia (and to a lesser extent, P . maltophilia) is usually isolated consistently, and can be associated with significant clinical deterioration . Occurrence of P . cepacia in CF respiratory tract cultures obtained close to the time of death rose nearly ten-fold from 1979 to 1982 . Strains representing all nonfermentative gram-negative species encountered were assayed for susceptibility to 17 newer antimicrobial agents . Ceftazidime, n-formimidoyl thienamycin, and aztreonam were most active; cefsulodin, ceforanide, and ceftriaxone were not active against these isolates.

J Hosp Infect, 1985 Mar, 6(1), 81 - 8
Prevention of bacterial contamination of water in dental units; Furuhashi M et al.; A study of the bacterial contamination of water in dental units was conducted in five dental practitioners' clinics and one university dental hospital in Tokyo . Various bacteria in numbers of 10(2)-10(5)/ml were detected in the water delivered from all 42 of the dental units examined, though very few bacteria were detected in the tap water upstream of the units . Most of the organisms isolated from the water were Gram-negative bacilli, predominantly Alcaligenes, Achromobacter and Pseudomonas spp . A new decontamination apparatus that could be incorporated into the pipe system of dental units was devised and its performance was evaluated in one unit . It effectively prevented bacterial contamination of the water.

Mikrobiologiia, 1985 Jan-Feb, 54(1), 108 - 13
{Nature of factors stimulating cobalamin production by Achromobacter cobalamini}; Shteinberg BI et al.; The object of this work was to study the nature of factors contained in molasses and maize extract and stimulating cobalaminogenesis in Achromobacter cobalamini . The activity of substrate fractions was analyzed to show that the stimulating substance was precipitated on the cation exchanger and eluted from it with HCl . The factor was found to be an organic nitrogen base readily soluble in water and ethanol but insoluble in ether, chloroform and methanol . It was stable upon heating in concentrated HCl . Betaine in the composition of molasses and choline in the composition of maize extract had similar properties . Their addition to the growth medium produced the same effect as that of molasses and maize extract . It is concluded therefore that cobalaminogenesis is stimulated in A . cobalamini by betaine in molasses and by choline in maize extract.

Appl Environ Microbiol, 1984 Dec, 48(6), 1214 - 20
In situ identification of bacterial species in marine microfouling films by using an immunofluorescence technique; Zambon JJ et al.; An immunofluorescence technique was developed for the in situ identification of specific bacteria in marine microfouling films . Microorganisms adherent to glass plates after 30 days of immersion in a synthetic seawater system were cultured and classified by biochemical tests, flagellar arrangement, and the API 20E system . All isolates were gram-negative aerobic or facultative motile rods, predominantly Pseudomonas spp . Rabbit antisera to the five dominant organisms including Achromobacter spp., Comamonas terrigena, P . putrefaciens, a yellow-pigmented Pseudomonas sp., and Vibrio alginolyticus were prepared . These antisera were shown to be species specific in indirect immunofluorescence assays against a battery of 26 marine isolates from 14 bacterial species, with the exception of antisera to the Pseudomonas spp, which cross-reacted with each other but not with test bacteria of other genera . These immunofluorescent reagents enabled the in situ identification of all five bacterial species in microfouling films . Low-surface-energy test plates had smaller numbers of adherent bacteria in microfouling films than medium-surface-energy test plates, suggesting that the degree of microfouling may be influenced by the surface energy . In addition, the reagents could identify up to 39% of the attached bacteria in microfouling films spontaneously formed on steel plates in flow cells deployed in different areas of the Atlantic Ocean . The microbial composition of the ocean-formed films varied with the geographical area of their formation . The present results indicate that immunofluorescence techniques may provide a rapid and reliable means to identify, in situ, specific bacteria in marine microfouling films.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1984 Dec, 179(6), 508 - 28
{Gaps in asepsis due to surgical caps, face masks, external surfaces of infusion bottles and sterile wrappers of disposable articles}; Graf W et al.; It is obvious that the surfaces of the boxes of sterile packed disposable instruments and infusion bottles are not sterile . The disposable surgical masks and surgical caps used for sterile clothing are delivered by the producers not sterile, either . To quantify these gaps and to judge their risks in the aseptic region the surfaces of 117 sterile packed disposable instruments and the inner sides of their boxes were examined bacteriologically . The surfaces of these objects proved to be not sterile by 21% and 4% were heavily contaminated with saprophytic germs . 3% of the examined articles showed pathogenic germs like Clostridium perfringens, Staphylococcus aureus, and Acinetobacter spec . 5-15% of the surfaces (glass- respectively plastic and labels) of the 331 infusion bottles proved to be heavily bacteriologically contaminated; 4-6% of them even showing pathogenic germs like Clostridium perfringens, Staphylococcus aureus, and Achromobacter spec . The surfaces of 25% of the examined disposable surgical masks and caps were considerably contaminated with saprophytic germs . Although pathogenic germs could not be detected, it means that these not sterile objects represent a considerable deficiency in surgical asepsis.

Br J Ophthalmol, 1984 Jul, 68(7), 472 - 4
Corneal ulcer due to Achromobacter xylosoxidans; Newman PE et al.; We report a case of corneal ulcer caused by the opportunistic organism Achromobacter xylosoxidans which developed during chronic topical steroid treatment of an eye with neovascular glaucoma . A . xylosoxidans has probably been underreported as a cause of ocular infection because of confusion between this organism and other Gram-negative organisms, particularly pseudomonas . A . xylosoxidans is resistant to aminoglycosides and some cephalosporins but not carbenicillin . This difference in antibiotic sensitivity patterns between A . xylosoxidans and pseudomonas makes an accurate differentiation between the 2 organisms important . This case was successfully treated after substituting topical carbenicillin for topical gentamicin and amikacin.

J Clin Microbiol, 1984 Jun, 19(6), 947 - 8
Bacteremia caused by Achromobacter species in an immunocompromised host; Kish MA et al.; A case of bacteremia caused by Achromobacter species in an immunocompromised patient is described . The patient responded to antibiotic therapy . Detailed antibiotic susceptibility data are presented.

Appl Environ Microbiol, 1984 May, 47(5), 947 - 51
Microbial biodegradation of 4-chlorobiphenyl, a model compound of chlorinated biphenyls; Masse R et al.; The biodegradation products of 4-chlorobiphenyl were analyzed in an Achromobacter sp . strain and a Bacillus brevis strain . Both strains generated the same metabolites, with 4-chlorobenzoic acid as the major metabolic product . Our results corroborate previous observations whereby most bacterial strains degrade the chlorobiphenyls via a major pathway which proceeds by an hydroxylation in position 2,3 and a meta-1,2 fission . However, we also detected several metabolites whose structure suggests the existence of other routes for the degradation of chlorinated biphenyls.

Anal Biochem, 1984 Apr, 138(1), 235 - 7
Effect of different collagenases on the isolation of viable hepatocytes from rat liver; Queral AE et al.; The isolation of viable hepatocytes from rat liver was found to depend on the source of collagenase (EC 3.4.24.3) more than any other single factor examined . Collagenase purified from Achromobacter iophagus/Bacillus polymyxa (collagenase/dispase) gave reproducibly high viability without the use of complex perfusion protocols.

Eur J Biochem, 1984 Feb 15, 139(1), 131 - 5
Ornithine-containing lipids in Thiobacillus A2 and Achromobacter sp; Thiele OW et al.; 20 bacterial strains (corresponding to 16 species) were screened for ornithine lipids . Only two species (Thiobacillus A2 and Achromobacter sp.) turned out to contain ornithine lipids (2.71 mmol/100 g and 0.38 mmol/100 g bacterial dry weight, respectively) . In both ornithine lipids, a 3-hydroxy fatty acid was amide-linked to the alpha-amino group of ornithine, a normal fatty acid was ester-linked to the 3-hydroxy group of the former . The predominant fatty acids were 18:1(11) and 3-hydroxy-20:1(13) in Thiobacillus A2, 16:0 and 3-hydroxy-18:1(11) in Achromobacter sp . All monounsaturated fatty acids (with one exception) belonged to the (n-7) family . 11, 12-Epoxy octadecanoic acid was identified among the ester-linked fatty acids of Thiobacillus A2 . Phosphatidylcholine was the principal phospholipid in both bacterial species.

J Clin Microbiol, 1984 Feb, 19(2), 140 - 3
Nosocomial colonization and infection by Achromobacter xylosoxidans; Reverdy ME et al.; Achromobacter xylosoxidans, a bacterial species named in 1971, is often isolated from aqueous environments, but little has been reported about its pathogenicity in humans, its epidemiological pattern, and its susceptibility to antibiotics and antiseptics . We were faced with an epidemic caused by this microorganism for 18 months in an intensive care unit . Two patients had fatal infections and 37 others were colonized . The source was the deionized water of the hemodialysis system . The 46 isolates were identified by comparison with the reference strain A . xylosoxidans ATCC 27061 . The characteristic cellular fatty acids of this species were demonstrated by gas-liquid chromatography . The minimal inhibitory concentrations of 27 antibiotics were determined . The isolates were susceptible to only two: moxalactam at 4 micrograms/ml and ceftazidime at 8 micrograms/ml . The minimal bactericidal concentrations of one disinfectant and three antiseptics were: sodium hypochloride, 109 micrograms/ml; chlorhexidine digluconate in ethanol solution, 15 to 125 micrograms/ml; polyvinylpyrrolidone iodine, 750 micrograms/ml; and iodine ethanol, 312 to 625 micrograms/ml.

Acta Microbiol Pol, 1984, 33(2), 163 - 70
Chitynolytic bacteria in water and bottom sediments of two lakes of different trophy; Donderski W; Chitynolytic bacteria were found in greater numbers in the eutrophic lake Jeziorak than in the mesotrophic lake Jasne . In both lakes higher chitynolytic activity was found in the planktonic than in the benthic bacteria . The capacity of chitin hydrolysis was found in bacteria of the genera Achromobacter, Bacillus, Nocardia, Pseudomonas, of the Flavobacterium-Cytophaga group and of the Enterobacteriaceae family . The chitinases synthesized by the studied bacteria were active in the pH range 5.0--8.0 . The majority of the strains tested showed maximum activity at pH 5.0 or 6.0.

Jpn J Antibiot, 1983 Dec, 36(12), 3399 - 404
{Chronological variations in blood isolates and their susceptibility to various antibiotics}; Fujita S et al.; Chronological variation in blood isolates obtained from 1968 to 1982 was studied . Escherichia coli was the most frequent isolates followed by Klebsiella species, glucose-non-fermentative Gram-negative rods (excluding Pseudomonas aeruginosa), Enterobacter-Serratia group, and Staphylococcus aureus . The isolation of glucose-non-fermentative Gram-negative rods, about half of which being Achromobacter xylosoxidans, increased greatly in number from 1980 . A total of 113 strains of blood isolates was examined for susceptibility to various antibiotics . Although there was a significant increase in S . aureus resistant to GM, these strains were susceptible to cephalosporins . In the 9 strains of GM-resistant aerobic Gram-negative rods (2 strains of Klebsiella, 2 strains of Enterobacter, 4 strains of Serratia and 1 strain of P . aeruginosa), 3 strains of Serratia species were also resistant to AMK . Two strains of LCM-resistant Bacteroides fragilis were susceptible to LMOX, but 1 strain was not inhibited by 100 micrograms/ml of CFX . Among the clinical specimens, differences were found in the rate of isolation of resistant strains, particularly in the case of Serratia and P . aeruginosa . Antibiotic resistant strains were isolated more frequently from urine and the least frequently from sputum . Since the frequency of isolation of resistant strains varied according to organism, clinical material and year of isolation, microbiological laboratories should develop their own data base from which clinicians can make rational therapeutic decisions.

J Clin Microbiol, 1983 Dec, 18(6), 1388 - 98
Examination of the morphology of bacteria adhering to peritoneal dialysis catheters by scanning and transmission electron microscopy; Marrie TJ et al.; We examined Tenckhoff peritoneal catheters by scanning and transmission electron microscopy to study the morphology of bacterial adherence . Two catheters were removed from uninfected patients, three from patients with exit site infections, four from patients with peritonitis, and one from a patient with both exit site infection and peritonitis . Infecting organisms included three of Staphylococcus aureus and one each of Enterobacter sp., Staphylococcus epidermidis, Achromobacter xylosoxidans, Serratia sp., Klebsiella sp., and Candida albicans . Considerable morphological variation in adherence to the peritoneal dialysis apparatus occurred . No inflammatory cells were ever seen in association with infected cuffs, only two of the five patients with peritonitis had inflammatory cells associated with their catheters . In both instances, these cells tended to occur in clumps and demonstrated no flattening when in contact with the surface . Colonization of the catheter was uneven--bacteria tended to occur in clusters . Extensive matrix formation was evident in several instances, and condensation of this matrix onto the bacteria during the dehydration process rendered clumps of bacterial cells amorphous at times . Bacteria were adherent to the subcutaneous cuff in those patients with exit site infections . Gram-negative bacteria attached to individual dacron fibers of the cuff, often several layers deep . Gram-positive bacteria tended to adhere in clusters.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1983 Sep, 177(6), 507 - 13
{Effect of human excrement on the air flora in the vicinity of moving trains}; Nikodemusz I et al.; The use of train lavatories by the passengers during their journey was investigated . Selective samples in Petri dishes were exposed for 1 or 2 minutes at different times against the direction of train motion . The analysis showed that enterobacteria (E . coli, Klebsiella, coliform bacteria, Proteus) and chromogenic bacteria (Pseudomonas, Achromobacter, Pseudomonas aeruginosa) can be verified in addition to yeast fungi, mould, actinomyces and micrococci . The colony count obtained depends on the climatic conditions, the passenger frequency as well as the site and period of exposure . The "air tunnel" formed by the travelling trains must be looked upon as contaminated . I would be advisable for the trains to travel with the windows locked . The intestinal bacteria are little risk to the environment but definitely injurious to the passengers themselves.

Am J Ophthalmol, 1983 Sep, 96(3), 354 - 61
Superinfections in herpes simplex keratitis; Boisjoly HM et al.; We reviewed 15 cases of culture-proven corneal superinfections in 15 patients (eight men and seven women ranging in age from 41 to 86 years) with recurrent herpes simplex keratitis . The factors that appeared to increase the risk of superinfection were the presence of an epithelial defect (found in all 15 cases), a history of recurrent herpetic keratouveitis (found in ten cases), and the use of topical corticosteroids (found in 13 cases) . Eight of the 15 patients were taking antibiotics at the time the superinfections were diagnosed, indicating that topical antibiotics do not provide sufficient protection . Gram-negative rods were found in six cases (Proteus mirabilis, Pseudomonas aeruginosa, Serratia marcescens, Klebsiella oxytoca, Enterobacter cloacae, and Achromobacter sp.) . Gram-positive organisms, often in association with another infecting agent, were found in six cases (Staphylococcus epidermidis, three cases; S . aureus, two cases; and Streptococcus sp., two cases) . Fungal superinfections were found in three cases (Cephalosporium acremonium, Candida albicans, and Aspergillus fumigatus, one case each) . Mycobacterium cheloni was found in two cases.

Biokhimiia, 1983 Jul, 48(7), 1172 - 80
{Bacterial formate dehydrogenase . Substrate specificity and kinetic mechanism of S-formyl glutathione oxidation}; Tishkov VI et al.; The substrate specificity of NAD-dependent formate dehydrogenase from the methylotrophic bacterium Achromobacter parvulus T1 was studied . The kinetic mechanism of S-formyl glutathione oxidation was determined . The initial velocity studies and inhibition analysis were carried out . It was shown that the kinetic mechanism for the enzyme with S-formyl glutathione as a substrate is similar to that with formate and is rapid-equilibrium random . Using independent methods, it was found that formate dehydrogenase forms a binary complex with S-formyl glutathione (Kd = 2.5 mM).

Jpn J Antibiot, 1983 Jul, 36(7), 1621 - 37
{Comparison of sensitivity patterns of bacteria isolated from clinical materials to the 2d generation cephamycins and cephalosporins and clinical application of these antibiotics}; Matsuo K et al.; In vitro susceptibilities of 1478 strains of various pathogens isolated from clinical materials in 1981 to 23 antibiotics were studied using Showa disk diffusion test . Prevalence of bacterial resistance to antibiotics was evaluated . New cephem antibiotics such as cefmetazole (CMZ), cefoxitin (CFX) and cefotiam (CTM) had no increased activity over old cephalosporins such as cephalothin (CET) and cefazolin (CEZ) against Gram-positive cocci . However, all the new ones showed greater activities, broadened spectra, and/or both against Gram-negative bacilli, each offering unique advantages . S . aureus: Susceptible strains (MICs less than 15 micrograms/ml) to CET, CEZ, CTM, CFX and CMZ were 95%, 69%, 92%, 88% and 99%, respectively . Prevalence of resistance to CEZ was greater than to other cephalosporins, showing bimodal distribution of MICs . This may be due to more use of this drug in the past . Sensitive strains to benzylpenicillin (PCG), ampicillin (ABPC), sulbenicillin (SBPC) and piperacillin (PIPC) were 22 to 63% . A striking bimodal distribution of MICs of these penicillins was characteristic . Sensitive strains to various aminoglycosides and tetracyclines were 52 approximately 93% and 97 approximately 100%, respectively . S . pyogenes: 83 approximately 100% of strains were susceptible to various penicillins, CET and CEZ, whereas susceptible strains to CTM, CFX and CMZ were 66 approximately 90% . S . pneumoniae: Almost all strains (99%) were susceptible to PCG at the level less than 0.2 units/ml (0.12 microgram/ml) . All strains were susceptible to various penicillins, cephalosporins, and cephamycins studied at the level less than 3 micrograms/ml . E . coli, K . pneumoniae and Proteus spp.: CTM, CFX and CMZ showed greater activity than CET and CEZ . Susceptible strains to the former antibiotics at the level less than 15 micrograms/ml were 93 approximately 94%, 82 approximately 95%, and 63 approximately 90%, respectively . Those to CET and CEZ were 61 approximately 83%, 85 approximately 92% and 53 approximately 58%, respectively . Among these cephalosporins, CMZ and CTM were the most effective, showing the least prevalence of resistance to these pathogens . H . influenzae: Susceptible strains (MICs less than 15 micrograms/ml) to ABPC, PIPC, CTM, CFX and CMZ were 80%, 81%, 84%, 55% and 71%, respectively . The majority of resistant strains to these beta-lactam antibiotics were sensitive to chloramphenicol, minocycline, doxycycline and erythromycin . P . aeruginosa, S . marcescens, E . aerogenes, Citrobacter and Achromobacter: CTM, CMZ and CFX were not effective against these pathogens.

Can J Microbiol, 1983 Jul, 29(7), 819 - 26
Comparative study of the beta-lactamase activity found in Achromobacter; Levesque R et al.; A survey of 21 clinical isolates of Achromobacter species demonstrated a high level of beta-lactamase activity in all strains tested . The beta-lactamases were characterized by isoelectric focusing, purification by affinity chromatography, determination of molecular weight, immunological identity, and genetic analysis . At least three distinct patterns of beta-lactamases were found in 19 strains . The kinetic values Km and Vmax measured by a microacidimetric method showed that all three types of enzymes are cephalosporinases and did not hydrolyse oxacillin, cloxacillin, and methicillin . Two of the three types of cephalosporinases studied, namely MULB 901 (isoelectric point (pI)7.4) and MULB 905(pI 9.3) are enzymes mediated by genes of chromosomal origin . The MULB 906 (pI 8.1) enzyme, however, which has been previously shown to be mediated by an 8.2 MDal nonconjugative plasmid, showed hydrolysis of cefoxitime, cefotaxin, and moxalactam by the bioassay . In all cases, beta-lactamase synthesis appeared constitutive . This study confirms that beta-lactamase activity is commonly found in Achromobacter and that these enzymes are different and of clinical interest when compared with those observed in other Gram-negative bacteria.

J Clin Microbiol, 1983 Feb, 17(2), 181 - 6
Serological classification of Achromobacter xylosoxidans; Shigeta S et al.; Adult rabbits were immunized with nine Achromobacter xylosoxidans strains by intravenous injection of Formalin-killed organisms . Antisera thus obtained were reciprocally titrated with the nine A . xylosoxidans strains, and seven sera were defined as serologically distinct . Three of nine antisera possessed one common antibody while also each having their own specific antibody . Ninety-five strains of A . xylosoxidans were examined for serotyping by a microtiter agglutination test with the nine antisera, and the strains were divided into seven serogroups . The distribution of the 95 strains among the serogroups was as follows: serogroup A, 15 strains; B, 17 strains; C, 43 strains; D, 2 strains; E, 4 strains; G, 8 strains; F, 2 strains . Four strains were not agglutinated with any of the antisera . Serogroups A, B, and F had the ability to reduce nitrate to nitrogen gas and to grow in heart infusion broth at 41 degrees C, although there were some exceptions . In contrast, most strains of serogroups C, D, and E could not produce nitrogen gas from nitrate, although they did produce nitrite . The strains of serogroup C could not grow at 41 degrees C, whereas those of serogroups D and E could . Thus, we concluded that serogroups A, B, and F referred to biotype IIIb described by Tatum, and serogroups C, D, and E referred to biotype IIIa . The serotyping of A . xylosoxidans may be useful for the analysis of nosocomial infections caused by these organisms.

Biochim Biophys Acta, 1983 Jan 5, 727(1), 115 - 21
Cell-surface collagen-binding protein in the procaryote Achromobacter iophagus; Keil-Dlouha V et al.; Collagen and its high-molecular-weight fragments specifically induce an extracellular collagenase (EC 3.4.24.8) in the Gram-negative Achromobacter iophagus . During the induction process the inducer is concentrated on the bacterial outer membrane . Two-dimensional electrophoresis of 125I-labelled outer membrane proteins has shown that, in particular, the amount of one protein which is already present on the surface of non-induced bacteria increases quantitatively when the inducer is added . After 125I-labelling of the cell membrane and its solubilization, the same protein is retained selectively on a gelatin-Sepharose column . It has isoelectric point of 4.9-5.1 and molecular weight of 40000 . This molecular weight is close to that of the 35000 of the collagenase subunit . However, their non-identity was proved in three independent ways: upon two-dimensional electrophoresis, only those proteins in the range corresponding to the collagenase dimer (Mr 70000-80000) react with fluorescent anticollagenase antibody system, whereas the spot of the collagen-binding protein (mr 40000) is negative; the solubilized collagen-binding protein is not retained by anticollagenase-Sepharose affinity chromatography; in vivo, it is not protected by anti-collagenase antibodies against lactoperoxidase iodination . A hypothesis for the possible role of the collagen-binding protein in the induction of collagenase is proposed.

Infection, 1983, 11 Suppl 1, S20 - 2
In vitro activity of new cephalosporins against multi-resistant gram-negative bacteria; Fainstein V et al.; The in vitro sensitivities of 47 multi-resistant gram-negative bacilli against six third generation cephalosporins were investigated . Overall, moxalactam was the most active compound and cefoperazone the least active . Ceftizoxime and ceftazidime were very active against Enterobacteriaceae . Ceftriaxone and ceftizoxime were active against Escherichia coli and Klebsiella spp., but were inactive against Pseudomonas spp.; Achromobacter sp . was resistant to all drugs, while Citrobacter freundii was sensitive to every antibiotic tested . These drugs may be clinically useful in infections with multi-resistant gram-negative bacteria.

Nauchnye Doki Vyss Shkoly Biol Nauki, 1983, (6), 83 - 7
{Action of polycarbon compounds on the oxidation of methanol and other Cl compounds by methylotrophic bacteria}; Zakharova EV; Cells of the facultative methylotrophic bacteria Achromobacter parvulus 1T, Pseudomonas sp AM1, Ps . methylica 2K, Ps . methylica 20T, Ps . oleovorans 52Z, Ps . fluorescens 45K exhibit the ability to oxidize methanol only on the medium with methanol . At the growth of various methylotrophic bacteria on the media containing both methanol and succinate their ability to methanol oxidation is lost, and oxidation of formaldehyde and formate occur at a low rate . Glucose, citrate, acetate and glycerol produce no repressive effect on the ability of the cells to oxidize methanol and other C1-compounds.

Zentralbl Mikrobiol, 1983, 138(5), 345 - 55
Cultivation of the bacterial strain Achromobacter delicatulus, producing an exocellular polyglucan-type polysaccharide in the fermentation tank FU-6; LasiK J et al.; Growth processes and biosynthesis of the exocellular polyglucan-type polysaccharide, produced by the bacteria Achromobacter delicatulus, were studied in the laboratory fermentation apparatus FU-6 under three completely different aeration systems . The purpose of this study was to find the most economical way of the polysaccharide biosynthesis . The growth rate and the synthesis of the exopolysaccharide were not limited either by the oxygen transfer or glucose content under the conditions examined . The best construction proved to be the fermentation tank with the lowspeed agitation system . In the case of the bacterial strain Achromobacter delicatulus, producing huge amounts of the thick slimy polyglucan exopolysaccharide, the industrial production may develop without any principal technical difficulties.

Connect Tissue Res, 1983, 11(2-3), 193 - 7
Solubilization and characterization of Chondrosia reniformis sponge collagen; Imhoff JM et al.; Chondrosia reniformis sponge collagen, insoluble in its native form, was solubilized by chemical modification of lysyl residues . The solubilized sponge collagen had the same amino acid composition as insoluble collagen and the helicoidal tertiary structure was found by negative Cotton effect to be the same as in native vertebrate collagens . Achromobacter iophagus collagenase, a collagen specific protease, hydrolyzed the soluble sponge collagen . These experiments confirmed that the protein had the same structure as collagen.

FEBS Lett, 1982 Dec 27, 150(2), 370 - 4
Purification and properties of alpha-amino-epsilon-caprolactam racemase from Achromobacter obae; Ahmed SA et al.; We have purified a unique enzyme, alpha-amino-epsilon-caprolactam racemase 945-fold from an extract of Achromobacter obae by Octyl-Sepharose CL-4B and Thiopropyl-Sepharose 6B and some other chromatographies . The purified enzyme was found homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and analytical ultracentrifugation . The enzyme has a monomeric structure with Mr approximately 50000 and a sedimentation coefficient (S20,w) of 4.28 S . The enzyme contains pyridoxal 5'-phosphate as a coenzyme . The pH optimum for the enzyme activity is approximately 9.0 . D- and L-alpha-amino-epsilon-caprolactams are the only substrates . The Km values for the D- and L-isomers are, 8 and 6 mM, respectively.

Biochim Biophys Acta, 1982 Dec 6, 709(1), 8 - 12
Chemical modification of lysine residues in bacterial formate dehydrogenase; Egorov AM et al.; Specific modification of 4.4 lysine residues per molecule of formate dehydrogenase, from the methylotrophic bacterium Achromobacter parvulus I by pyridoxal, results in complete inactivation of the enzyme . The concentration effect of the modifying agent and substrates on the inactivation of formate dehydrogenase has been studied . Coenzymes do not protect the enzyme from inactivation . Complete maintenance of enzyme activity was achieved in the presence of saturating concentrations of the formate and upon formation of the ternary complex, enzyme-NAD-azide . Formate specifically protects two lysine residues per dimer molecule of the enzyme from modification . The presence of one essential lysine residue in the substrate-binding region of the enzyme active site is assumed.

J Hyg (Lond), 1982 Dec, 89(3), 529 - 38
Characterization of the major antigens of Haemophilus equigenitalis (contagious equine metritis organism); Corbel MJ et al.; Immunoelectrophoresis of ultrasonically disrupted Haemophilus equigenitalis (contagious equine metritis organism) cells against rabbit and equine antisera disclosed at least 11 precipitating antigens . Two of these, a polysaccharide and a lipopolysaccharide-protein complex, were of high molecular weight and located on the cell surface . The remaining antigens were intracellular and were small- to medium-sized proteins . The surface antigens were the most significant in relation to the serological response in infected horses . They also reacted with sera from apparently healthy cattle, but the reason for this was not determined . No serological cross-reaction between H . equigenitalis and species of Achromobacter and Moraxella was detected.

Mikrobiologiia, 1982 Nov-Dec, 51(6), 910 - 4
{Effect of organic substances on the growth and cobalamin genesis of Achromobacter cobalamini}; Shteinberg BI et al.; Corn steep and molasses were shown to stimulate cobalaminogenesis in Achromobacter cobalamini . Their concentrations optimal for the culture growth and vitamin B12 synthesis were found . Corn steep at a concentration of 2% accelerated the growth . The biomass yield and the content of B12 were maximal after 48 h of fermentation . In the medium with molasses, the growth was stimulated at 1% of molasses, and the vitamin biosynthesis at 5% . When 5% of molasses was added into the mineral medium, the rate of biosynthesis increased by a factor of 5 to 7 as compared to the control . A 5% aqueous solution of molasses containing 1.1 g/l of potassium phosphates produced the same effect as an addition of molasses to the complete mineral medium.

Antimicrob Agents Chemother, 1982 Nov, 22(5), 832 - 8
In vitro activity of azthreonam, a monobactam antibiotic; Jacobus NV et al.; We studied the activity of azthreonam (SQ 26,776), a novel monocyclic beta-lactam compound, against a variety of clinical isolates . It was more potent than moxalactam, cefoperazone, cefamandole, cefoxitin, ticarcillin, tobramycin, or amikacin against strains of Klebsiella spp., Serratia spp., and the Proteus group . It was highly effective against Escherichia coli and strains of Salmonella spp . The median minimal inhibitory concentration for all species of Enterobacteriaceae was less than or equal to 2 micrograms/ml . Azthreonam was moderately active against Pseudomonas aeruginosa, including tobramycin-resistant strains, and against Pseudomonas cepacia (median minimal inhibitory concentration, 16 to 32 micrograms/ml), but was weakly active against Pseudomonas maltophilia and strains of Acinetobacter spp . and Achromobacter spp . The drug showed little activity against Staphylococcus aureus, enterococci, and anaerobic bacteria, including Bacteroides fragilis, Clostridium spp., and gram-positive cocci . Like moxalactam and cefoperazone, azthreonam exhibited a considerable inoculum effect with strains of Enterobacter spp . and Pseudomonas spp . Combination with clavulanic acid did not increase the activity of azthreonam against S . aureus but was synergistic for 5 of 15 strains of B . fragilis . Azthreonam is about 50% bound to human serum protein . The selective range of activity of this compound could be of clinical benefit.

J Clin Microbiol, 1982 Jun, 15(6), 1144 - 7
Growth of non-Campylobacter, oxidase-positive bacteria on selective Campylobacter agar; Moskowitz LB et al.; A total of 67 oxidase-positive, gram-negative bacteria were tested for growth on selective Campylobacter agar (Blaser formulation, BBL Microbiology Systems, Cockeysville, Md.) at 42 degrees C under microaerophilic conditions . Although the growth of most of these bacteria was prevented, all strains of Achromobacter xylosoxidans, Pseudomonas aeruginosa, Pseudomonas putrefaciens, Pseudomonas alcaligenes, and Pseudomonas pseudoalcaligenes grew as well as Campylobacter fetus subsp . jejuni.

Sem Hop, 1982 May 6, 58(18), 1129 - 33
{Study of the spermatic bacterial flora in infertile males (author's transl)}; Granouillet R et al.; Two groups of infertile males (65 and 132 patients) have been investigated in two different laboratories, with two different methods to obtain semen . The bacteriological results are quite similar in the two groups . The microorganisms which have been isolated are : beta-hemolytic Streptococcus, Proteus, Epidermidis staphylococcus, Micrococcus, Corynebacter, Viridans streptococcus, Klebsiella, Pseudomonas, Enterobacter, Bacillus, Neisseria, Escherichia coli, anaerobic Staphylococcus, anaerobic Streptococcus, anaerobic Corynebacter type IV . Fungus, Achromobacter, 20 p . cent of the patients are chronically infected without any clinical signs . This infection is probably of prostatic origin with an important number of bacteria in the semen (more than 3 000/ml) . No relation has been shown between the bacteriological data and the physical and cytological characteristics of the sperm, except the pH : semens with a low pH are generally azoospermic and highly contaminated.

Am J Epidemiol, 1982 May, 115(5), 785 - 93
An outbreak of Achromobacter xylosoxidans related to diagnostic tracer procedures; McGuckin MB et al.; In December, 1978, an investigation was undertaken to determine the source of infection in five patients in one hospital with hospital-associated bacteremia due to Achromobacter xylosoxidans . Review of their records showed that each had a diagnostic tracer procedure preceding the bacteremia and that no other procedures were common to all . Further investigation revealed that patients from three other hospitals were studied using diagnostic tracer materials from the index hospital . Five patients with confirmed A . xylosoxidans bacteremia and four suspected cases were identified in these hospitals, and all had a scan before the bacteremia was detected . No other A . xylosoxidans isolates were identified in any of the hospitals in the preceding two years . Although not confirmed, the source appeared to be stored non-bacteriostatic saline . Effective control measures included a sterility testing program and use of pre-packaged single dose vials of saline . Diagnostic tracer studies should be added to the list of procedures known to cause hospital-acquired bacteremias.

J Infect Dis, 1982 May, 145(5), 753 - 61
A plasmid-mediated cephalosporinase from Achromobacter species; Levesque R et al.; An unusual cephalosporinase in Achromobacter species was characterized biochemically; the enzyme had a pI of 8.1 and a molecular mass of 36,200 daltons, and it was not inhibited by p-chloromercuribenzoate or cloxacillin . Specific antiserum neutralized enzymatic activity . Agarose gel electrophoresis of the DNA of two strains (MULB 906 and MULB 912) revealed at least three plasmid bands; cured strains demonstrated a simultaneous loss of beta-lactamase and plasmid DNA . Resistance to beta-lactam antibiotics was transferred by transformation of Escherichia coli strain HB101 with plasmid DNA . This plasmid-mediated beta-lactamase differed from the two types of chromosomal cephalosporinases (pI 7.4 and 9.3, respectively) found in a survey of clinical isolates of Achromobacter species . This enzyme also differed in its biochemical properties from all of the other known plasmid-mediated beta-lactamases.

Gene, 1982 Apr, 18(1), 69 - 75
Mapping of the plasmid (pLQ3) from Achromobacter and cloning of its cephalosporinase gene in Escherichia coli; Levesque R et al.; We have constructed a physical map of the plasmid pLQ3 which was originally isolated from Achromobacter and which codes for a beta-lactamase . The enzyme specified by pLQ3 is expressed in Escherichia coli and is unusual in that it is a cephalosporinase, an enzyme usually coded for by chromosome . Plasmid pLQ3 is 12.4 kb in length and has a unique Bam HI site and two BglII sites . From a BamHI + BglII double digest of pLQ3, we have constructed a "shortened" plasmid, pLQ10, in which a 2.96-kb fragment is deleted . We have constructed a clone, pLQ22, in which a 3.27-kb fragment of pLQ3, carrying the beta-lactamase gene, is inserted into the BamHI site of pACYC184 . By "comparative mapping" of single and multiple digests of each of these plasmids, we have been able to locate the cleavage sites for PstI, which makes seven cuts in pLQ3.

Tohoku J Exp Med, 1982 Mar, 136(3), 251 - 61
Experimental infection of immunocompromised mice with Achromobacter xylosoxidans; Hyodo S et al.; The natural resistance of mice to Achromobacter (A.) xylosoxidans was decreased to 1/80 by a single dose of 250 mg of cylophosphamide (CY) per kg intraperitoneally . When mice were infected with 1 X 10(9) A . xylosoxidans, the numbers of organisms in the liver, spleen, kidneys, lungs and heart blood reached to more than 10(6) per g at 6 hr after infection, and then decreased to less than 10(4) at 96 hr after infection . However, in the CY-treated mice, the numbers of organisms in these organs began to increase rapidly at 6 hr after infection and exceeded 10(9) per g at 48 hr when the mice died . Seventeen of 18 immunocompromised mice and 23 of 24 normal mice which were infected with 1 to 2 X 10(9) organisms of A . xylosoxidans, respectively, were still living until 4 weeks after infection when they were killed and examined . Thirteen of the 33 surviving mice which were examined for bacteriological assay were chronically infected with A . xylosoxidans, some in the spleen, others in the kidney, lung, brain and/or heart blood but none were infected in the liver . Most of the infected and surviving mice had produced an antibody against A . xylosoxidans and its titer was higher in the bacteriologically "cured" group than in the chronically infected one . All surviving mice were resistant to the 2nd challenge with a lethal dose of A . xylosoxidans even when they received an effective dose of CY . The surviving mice showed activated cellular immunity and rather depressed humoral immunity as compared with normal mice or the mice which were infected with killed A . xylosoxidans.

Pathol Biol (Paris), 1982 Jan, 30(1), 22 - 6
{Study of the spermatic bacterial flora in infertile males (author's transl)}; Granouillet R et al.; Two groups of infertile males (65 and 132 patients) have been investigated in two different laboratories, with two different methods to obtain semen . The bacteriological results are quite similar in the two groups . The microorganisms which have been isolated are : beta-hemolytic Streptococcus, Proteus, Epidermidis staphylococcus, Micrococcus, Corynebacter, Viridans streptococcus, Klebsiella, Pseudomonas, Enterobacter, Bacillus, Neisseria, Escherichia coli, anaerobic Staphylococcus, anaerobic Streptococcus, anaerobic Corynebacter type IV, Fungus, Achromobacter . 20 p . cent of the patients are chronically infected without any clinical signs . This infection is probably of prostatic origin with an important number of bacteria in the semen (more than 3,000/ml) . No relation has been shown between the bacteriological data and the physical and cytological characteristics of the sperm, except the pH : semens with a low pH are generally azoospermic and highly contaminated.

Vet Med (Praha), 1982 Jan, 27(1), 25 - 9
{Lipolytic microorganisms in liver products}; Lukasova J et al.; Products from liver were studied for the content of lipolytic microorganisms during production and storage . Lipolytic microorganisms were found in all samples of the products . Their occurrence ranged from 10(4) to 10(7) organisms per g . In finished products and in products stored at a refrigerator temperature their numbers were much lower (10-10(4) per g) . Some samples were free from them . The isolated colonies of lipolytic microorganisms were included in the genera Bacillus, Staphylococcus, Pseudomonas, Achromobacter . The acidity number was in correlation with the number of lipolytes in products before thermal processing and in stored products.

J Clin Microbiol, 1982 Jan, 15(1), 43 - 7
Accuracy of a rapid carbohydrate oxidation microtube method for identification of nonfermentative gram-negative bacilli; Qadri SM et al.; A rapid carbohydrate oxidation microtube system (Carr Microbiologicals, Wichita, Kans.), designed for detecting the saccharolytic activity of gram-negative, nonfermenting bacilli, was evaluated and compared with the conventional oxidation-fermentation method . The oxidation of glucose, maltose, lactose, and xylose was tested with 430 strains of Pseudomonas, Acinetobacter, Achromobacter, Alcaligenes, Moraxella, Flavobacterium, and Bordetella species . More than 95% of the isolates tested gave correct oxidation reactions within 4 h in the rapid carbohydrate oxidation microtubes, whereas oxidation-fermentation media required 24 h to achieve the same sensitivity . The microtube system was found to be simple, accurate, rapid, and economical.

Acta Microbiol Pol, 1982, 31(3-4), 293 - 9
Studies on pectolytic bacteria in water and bottom sediments of two lakes of different trophy; Donderski W; Pectolytic bacteria were found in greater numbers in the eutrophic than in the mesotrophic lake . However, higher pectolytic activity was exhibited by the bacteria derived from the mesotrophic lake . In the eutrophic lake higher endo- and exo-PG activity was found in the benthic than in the panktonic bacteria, whereas in the mesotrophic one higher endo-PG activity was exhibited by the planktonic and higher exo-PG activity by the benthic bacteria . The polygalacturonases synthesized by the bacteria of the genera Achromobacter, Bacillus, Pseudomonas, of the group Flavobacterium - Cytophaga and of the family Enterobacteriaceae were active in the pH range 6.0-9.0 . The majority of tested strains showed maximum activity at a temperature of 30 degrees C.

Cancer Treat Rep, 1981 Nov-Dec, 65(11-12), 1077 - 81
Immunosuppressive properties and circulating life of Achromobacter glutaminase-asparaginase covalently attached to polyethylene glycol in man; Abuchowski A et al.; The immunosuppressive effects and circulating life of Achromobacter glutaminase-asparaginase (GA) covalently attached to polyethylene glycol (PEG) were examined in human subjects following a single iv dose of 1000 IU/m2 . Plasma half-life of PEG-GA was 72 hours . Skin test reactivity to recall antigens (mumps and tuberculin) was lost in all four patients tested . In vitro phytohemagglutinin-induced blastogenesis, "natural killing," and phytohemagglutinin-induced cell cytotoxicity was diminished as long as enzyme levels were detectable . In vivo and in vitro activities returned to normal following total plasma clearance of enzyme.

Biochim Biophys Acta, 1981 Oct 12, 677(2), 220 - 4
Improvement in the persistence of microbial asparaginase and glutaminase in the circulation of the rat by chemical modifications; Blazek R et al.; Three enzymes used in cancer chemotherapy (asparaginases from Escherichia coli and Erwinia carotovora and glutaminase from Achromobacter) were each reacted with four amino specific reagents (ethyl acetimidate, O-methylisourea, succinic anhydride, and formaldehyde/sodium borohydride) . The half-lives of the modified enzymes measured in the blood of rats showed that guanidation, acetimidation and reductive alkylation were more likely to increase the persistence of the native enzymes than succinylation . However, the improvement in the persistence of any one enzyme after any one modification could not be predicted from the results with the others . It was concluded that changes in persistence caused by each modification were due to the different effects on the tertiary structure of each native enzyme . The advantages of chemical modification for increasing the persistence of enzymes over other methods such as encapsulation or aggregation are discussed.

Biochim Biophys Acta, 1981 Jul 24, 660(1), 44 - 50
Studies on a new proteolytic enzyme from A chromobacter lyticus M497-1 . I . Purification and some enzymatic properties; Masaki T et al.; Achromobacter lyticus M497-1 produces three kinds of alkaline proteases (protease I, II and III) in culture medium along with the bacteriolytic enzyme (Masaki, T., Nakamura, K., Isono, M . and Soejima, M . (1978) Agric . Biol . Chem . 42, 1443--1445) . Among these three proteases, Achromobacter protease I (EC 3.4.21.-) shows strict splitting for lysine residues at the carboxyl side of the splitting point . This enzyme was purified through a sequence of benzalkonium chloride treatment, acetone fractionation, CM-cellulose and DEAE-cellulose treatment chromatography on AH-Sepharose 4B and isoelectric focusing method . This form was shown to be homogeneous by polyacrylamide gel electrophoresis and ultracentrifugation analysis . The physicochemical properties of the enzyme were: Mr 30 500; partial specific volume (v), 0.717 ml/g; intrinsic viscosity (nu), 0.0385) dl/g; isoelectric point (pI) 6.9; and E1%1cm at 280 nm, 18.77 . The enzyme was composed of 294 residues of amino acid per molecule, with glycine as NH2-terminal and lysine as COOH-terminal amino acids . The optimum pH values with casein, Bz-lys-pNA and Tos-Lys-OMe were 8.5--10.7, 9.0--9.5 and 7.8--8.2, respectively . The enzyme was inhibited by iPr2P-F, PhCH2SO2F and Tos-LysCH2Cl but not by Tos-ArgCH2Cl, EDTA, o-phenanthroline and PCMB.

Biochim Biophys Acta, 1981 Jul 24, 660(1), 51 - 5
Studies on a new proteolytic enzyme from Achromobacter lyticus M497-1 . II . specificity and inhibition studies of Achromobacter protease I; Masaki T et al.; The unique specificity of Achromobacter protease I for lysine residue was investigated using synthetic and natural substrates, i.e., lysine derivatives, arginine derivatives, lysine vasopressin, substance P, ACTH and insulin . The enzyme cleaved only the -Lys-X- bonds in the above substrates . The binding affinity of alkylamines as determined by Ki was much stronger than that of the corresponding alkylguanidines.

Biochim Biophys Acta, 1981 Jun 15, 659(2), 283 - 91
Chemical characterization of the homogeneous collagenase from Clostridium histolyticum; Emod I et al.; Pure collagenase (clostridiopeptidase A, EC 3.4.24.3) having a molecular weight of 70 000 was obtained from the culture medium of Clostridium histolyticym by a combination of ultrafiltrations, molecular sieve, affinity and hydrophobic chromatography . The value of its specific activity is the highest of those described previously but 6-times lower than that of the collagenase from Achromobacter iophagus (EC 3.4.24.8) . Its amino acid composition differs from previous data, namely by the presence of cysteine, methionine, tryptophan and O-phosphoserine residues . In contrast to Achromobacter collagenase it does not dissociate in subunits during the deactivation by EDTA or LiCl/glycine buffer at pH 10.5 . Existence of multiple forms of Clostridium collagenase previously described is discussed as being due to autolysis of a single molecular species or to a different degree of phosphorylation.

Biochim Biophys Acta, 1981 May 14, 659(1), 141 - 9
Study of the role of arginine residues in bacterial formate dehydrogenase; Egorov AM et al.; Modification of 12 arginine residues per molecule of formate dehydrogenase (formate : NAD+ oxidoreductase, EC 1.2.1.2.) from the methylotrophic bacterium, Achromobacter parvulus I, by 2,3-butanedione results in complete inactivation of the enzyme . Inactivation of the enzyme is reversible and proceeds in two steps via formation of the intermediate enzyme-butanedione complex . Coenzymes but not formate effectively protect formate dehydrogenase from inactivation . Complete maintenance of enzyme activity and specific protection of one arginine residue per enzyme subunit are achieved on formation of the binary complex, enzyme-NAD, or the ternary complex, enzyme-NAD-azide . One arginine residue is supposed to be located at the NAD-binding site of the formate dehydrogenase active centre.

Zentralbl Bakteriol A, 1981 Feb, 248(4), 509 - 25
{The identification of nonfermentative gram-negative bacteria . Experiences with 676 apyocyaninogenic strains (author's transl)}; Berger U et al.; During a period of 16 months 1757 strains of nonfermentative gram-negative rods have been isolated from clinical material . Of the, 1205 (69%) were P . aeruginosa, 124 (10%) of which failed to produce pyocyanin . The apyocyaninogenic strains as well as the remaining 552 isolates were differentiated by steps according to a diagnostic scheme developed by us . For identification of species two or three steps were needed . By this procedure, 530 of the 552 strains could be assigned to nineteen species within the genera Pseudomonas, Achromobacter, Alcaligenes, Flavobacterium, Agrobacterium and Acinetobacter . 17 strains could not be identified below the genus level, one strain belonged to CDC-group VE-2 and four strains were not identifiable . 72% of the 552 strains belonged to only four species: Pseudomonas putida, P . maltophilia, Acinetobacter lwoffii and A . anitratus.

Biochim Biophys Acta, 1980 Oct, 615(2), 436 - 48
Chemical modifications of Achromobacter collagenase and their influence on the enzymic activity; Trocheris I et al.; A study of the influence of chemical modifications on the activity of Achromobacter iophagus collagenase (EC 3.4.24.8) has led to the following conclusions: a modification of 4 out of 80 COOH groups with carbodiimide led to 90% loss of enzymic activity . A 70% inactivation was found after modification of two tyrosines out of 30 with tetranitromethane . The modification of four to six tryptophans out of 16 with 2-hydroxy-5-nitrobenzyl bromide decreased enzyme activity to 36% . This inactivation is accelerated in the presence of collagen . An increase of reagent/enzyme molar ratio led to a modification of 16 tryptophan residues and denaturation of Acahromobacter collagenase . A modification of two arginines out of 18 with 1,2-cyclohexanedione and eight NH2 groups out of 24 with 2,3-dimethyl maleic anhydride does not change the collagenolytic activity . All NH2 groups become available for 2,3-dimethyl maleic anhydride after dissociation of the dimer . A possible analogy of hydrolytic site of collagenase with that of two other known bacterial metalloproteinases (thermolysin and Bacillus subtilis neutral proteinase (EC 3.4.24.4)) is discussed.

Antimicrob Agents Chemother, 1980 Sep, 18(3), 483 - 6
In vitro activity of cefoperazone against nonfermenters and Aeromonas hydrophila; Fass RJ; The in vitro activity of cefoperazone against 380 strains (33 species and unnamed groups) of nonferenters and 20 strains of Aeromonas hydrophila was studied by a microdilution method of determining minimal inhibitory concentrations . For comparison, the activities of ampicillin, ticarcillin, and cefamandole were simultaneously studied . Cefoperazone was the most active drug against Pseudomonas sp., Achromobacter xylosoxidans, Flavobacterium meningosepticum . Flavobacterium sp . (group IIb), Group IVc biotype 2, Group Ve, and Aeromonas hydrophila . Against other organisms it was equivalent in activity to or less active than comparative antibiotics . The only organisms which were usually resistant to 128 microgram of cefoperazone per ml were strains of Flavobacterium odoratum and Group IIk biotype 1.

J Clin Microbiol, 1980 Aug, 12(2), 282 - 3
Pancreatic abscess associated with Achromobacter group Vd biovar 1; Appelbaum PC et al.; A case of pancreatic abscess associated with Achromobacter biovar 1 in a 75-year-old man with multiple predisposing debilitating conditions is presented . The infection responded to antibiotic therapy, but the patient died from unrelated causes.

Biochim Biophys Acta, 1980 Jul 24, 624(1), 51 - 9
Circular dichroism comparative studies of two bacterial collagenases and thermolysin; Heindl MC et al.; The recently isolated and purified collagenase produced by Achromobacter iophagus, the collagenase from Clostridium histolyticum, and thermolysin, three enzymes having common properties, were studied by circular dichroism . From the spectra of the aqueous solutions obtained in the peptide region, the fraction of alpha helix, beta sheet and aperiodic segments in the three proteins could be estimated . Good similarity was found between Achromobacter collagenase and thermolysin, which both contain a high fraction of alpha helix . Side-chain contributions were analyzed in the aromatic region of thespectra: effects of pH and of organic solvents were observed, showing the strong influence of surroundings on the stabilization of the proteins.

Mikrobiologiia, 1980 Jul-Aug, 49(4), 621 - 3
{Preservation of lyophilized cultures of saprophytic microorganisms}; Kupletskaia MB et al.; All of the tested saprophytic microbial cultures remained viable upon 11--16 years of storage in the lyophilized state at 3--6 degrees . The cultures belonged to the following taxonomic groups: Acetobacter, Achromobacter, Azotobacter, Bacillus, Chromatium, Escherichia, Flavobacterium, Lactobacillus, Micrococcus, Proteus, Propionibacterium, Pseudomonas, Rhizobium, Sarcina, Serratia, Streptococcus, Torula, Mycobacterium, Actinomyces . The number of viable cells in the tubes decredased only slightly within this time and remained at the same level as immediately after lyophilization.

J Biochem (Tokyo), 1980 May, 87(5), 1395 - 402
L-Lysine: 2-oxoglutarate 6-aminotransferase . Subunit structure composed of non-identical polypeptides and pyridoxal 5'-phosphate-binding subunit; Yagi T et al.; L-Lysine:2-oxoglutarate 6-aminotransferase from Flavobacterium lutescence (= Achromobacter liquidum) has been shown to be composed of one each of four non-identical subunits, A, B1, B2, and C . The subunits were isolated by gel filtration, and DEAE-cellulose chromatography in the presence of 8 M urea . Their molecular weights were determined by ultracentrifugation, gel electrophoresis and gel filtration: subunit A 24,000; B1 28,000; B2 28,000; C 45,000 . These subunits were all different in amino acid composition . Of the two molecules of bound pyridoxal 5'-phosphate, the one which absorbs at 415 nm is bound to subunit B2 and participates in the catalytic action of the enzyme.

Biokhimiia, 1980 May, 45(5), 854 - 63
{Two ways of formate oxidation in methylotrophic bacteria}; Rodionov IuV et al.; The cells of Achromobacter parvulus, strain 1T, when grown in a methanol-containing medium, have two formate dehydrogenases, i.e . NAD-linked formate dehydrogenase and the formate dehydrogenase reducing the ferricyanide and tetrazolia . Only the latter enzyme was found in the cells grown in a medium with glycerol as a carbon source . These enzymes differ with respect to Km for formate and antigenic specificity . Km for formate oxidation by the cells of A . parvulus is lower than for formate of the NAD-dependent formate dehydrogenase and is equal to Km for the enzyme reducing artificial electron acceptors . The results obtained are discussed in terms of the existence of two cytochrome oxidase systems in the methylotrophic bacteria, differing in their sensitivities to the inhibition by formate.

Mikrobiologiia, 1980 Mar-Apr, 49(2), 215 - 20
{Respiratory resistance of methylotrophic bacteria to formate and cyanide}; Zakharova EV et al.; Whole cells and cell-free preparations of the methylotrophic bacteria, Pseudomonas sp . AM 1 and Achromobacter parvulus, can oxidize formate at tis concentration in the reaction medium up to 1 M . The respiration of whole cells is registered at a concentration of formate greater than 10(-2) M, while that of cell-free extracts at a formate concentration greater than 5 X 10(-5) M . This seems to be due to the presence of a permeability barrier in cells for formate . The oxidation of reduced TMPD and exogenous cytochrome c by the membrane preparations of the two bacteria is inhibited by formate and cyanide; Ki50% = 2.5 X 10(-2) and 10(-6) M, respectively . The oxidation of NADH by the membrane preparations of the bacteria is not inhibited by 1 M formate and 5 X 10(-4) M cyanide but is inhibited by formaldehyde with Ki50% = 3 X 10(-2) M . Formaldehyde has no effect on the oxidation of reduced TMPD and cytochrome c at concentrations greater than 2 X 10(-1) M . These data indicate that respiration of the studied methylotrophic bacteria in the presence of high formate concentrations should be attributed in the presence of a branched electron transport chain in them; one branch of the chain is resistant to formate and cyanide, but is sensitive to formaldehyde.

J Clin Microbiol, 1980 Feb, 11(2), 141 - 5
Clinical and laboratory characteristics of Achromobacter xylosoxidans infection; Igra-Siegman Y et al.; Achromobacter xylosoxidans was isolated from six patients . The organism causes opportunistic infections in patients who are compromised . A . xylosoxidans is a catalase- and oxidase-positive, motile, gram-negative rod that oxidizes xylose and glucose . The organism exists in a water environment and may be confused with Pseudomonas species . Unlike pseudomonas, achromobacter has peritrichous flagella . The clinical and laboratory characteristics of A . xylosoxidans are presented.

CRC Crit Rev Biochem, 1979 Dec, 7(2), 103 - 19
New experiments of biotin enzymes; Lynen F; The objects of structural studies on biotin-enzymes were acetyl CoA-carboxylase and pyruvate carboxylase of Saccharomyces cerevisiae and beta-methylcrotonyl CoA-carboxylase and acetyl CoA-carboxylase of Achromobacter IV S . It was found that these enzymes can be arranged in three groups . In the first group, as represented by acetyl CoA-carboxylase of Achromobacter, the active enzyme could be resolved in three types of functional components: (1) the biotin-carboxyl carrier protein, (2) the biotin carboxylase, and (3) the carboxyl transferase . In the second group, as represented by beta-methylcrotonyl CoA-carboxylase from Achromobacter only two types of polypeptides are present . The one carries the biotin carboxylase activity together with the biotin-carboxyl-carrier protein, the other one carries the carboxyl transferase activity . In this third group, as represented by the two enzymes of yeast, all three catalytic functions are incorporated in one multifunctional polypeptide chain . The evolution of the different enzymes is discussed . The animal tissues acetyl CoA-carboxylase is under metabolic control, as known from previous studies . It thus has to be expected that the levels of malonyl CoA in livers of rats in all states of depressed fatty acid synthesis are much lower than under normal conditions because the carboxylation of acetyl CoA is strongly reduced and cannot keep pace with the consumption of malonyl CoA by fatty acid synthetase . A new highly sensitive assay method for malonyl CoA was developed which uses tritiated NADPH and measures the incorporation of radioactivity into the fatty acids formed from malonyl CoA in the presence of purified fatty acid synthetase . The application of this method to liver extracts showed that the level of malonyl CoA which amounts to about 7 nmoles per gram of wet liver drops to less than 10% within a starvation period of 24 hr and even further if the starvation period is extended to 48 hr . A low malonyl CoA concentration is also found in the alloxan diabetic animals and in animals being fed a fatty diet after starvation . On the other hand, feeding a carbohydrate rich diet leads to malonyl CoA levels surpassing the levels found after feeding a balanced diet . These observations reconfirm the concept that fatty acid synthesis is principally regulated by the carboxylation of acetyl CoA.

Mikrobiyol Bul, 1979 Oct, 13(4), 389 - 91
{An Achromobacter xylosoxidans strain isolated from pleural empyema (author's transl)}; Saygun N et al.; The isolation of an Achromobacter xylosoxidans strain from a 32 year old man with pleural empyema, and its biochemical characteristics are reported.

J Clin Microbiol, 1979 Oct, 10(4), 525 - 8
Semiquantitative catalase test as an aid in identification of oxidative and nonsaccharolytic gram-negative bacteria; Chester B; A simple and rapid semiquantitative slide catalase test useful for the identification of oxidative and nonsaccharolytic gram-negative bacteria, i.e., "nonfermenters," is described . Using the interpretative criterion of time of appearance of oxygen bubbles in 3% hydrogen peroxide, three categories of nonfermenters were established . The rapid catalase producers included Achromobacter xylosoxidans and Achromobacter species; Acinetobacter anitratus and Acinetobacter lwoffii; Bordetella bronchiseptica; CDC group IVE; Pseudomonas aeruginosa, P . fluorescens, P . putida, P . diminuta, and P . acidovorans; and Moraxella urethralis and M-6 . The delayed catalase producers included Bordetella parapertussis, CDC group VA-1, P . alcaligenes, P . cepacia, P . mendocina, P . pickettii (VA-2), P . pseudoalcaligenes, P . putrefaciens, P . stutzeri, P . testosteroni, and P . vesicularis . The third group consisted of an additional 17 taxa of nonfermenters which were classified as moderate catalase producers.

Biochemistry, 1979 Sep 4, 18(18), 3917 - 20
Studies with mechanism-based inactivators of lysine epsilon-transaminase from Achromobacter liquidum; Shannon P et al.; Analogues of lysine containing a 4,5-acetylenic linkage (lysyne) or a cis- or trans-4,5-olefinic linkage (lysenes) function as substrates for a homogeneous L-lysine epsilon-transaminase from Achromobacter liquidum but partition between transamination and time-dependent inactivation . The partition ratio is lowest for lysyne (40 per inactivation event) and higher for trans-lysene (160 per inactivation event), and the cis-lysene transaminates 1600 times per inactivation event . cis-Lysene yields alpha-picolinate as a detectable accumulating product, presumably from cyclization of initial 6-aldehyde to dihydropicolinate and spontaneous autoxidation . The trans isomer also yields some picolinate as an identifiable product . The product from the few lysyne turnovers is as yet unknown but has strong absorbance at 318 nm . The inactive enzyme species from all three lysine analogues slowly (overnight) regain full activity after gel filtration chromatography and dialysis, suggesting reversal of the initial adduct-forming reaction . Initial studies with partially purified pseudomonad lysine alpha-racemase show alpha-3H incorporation from 3H2O but no inactivation consistent with the expectation that these lysine analogues could act readily as mechanism-based inactivators for pyridoxal P enzymes which act at the epsilon- but not the alpha-carbon of lysine.

Appl Environ Microbiol, 1979 Sep, 38(3), 359 - 64
Stimulation of L-asparate beta-decarboxylase formation by L-glutamate in Pseudomonas dacunhae and Improved production of L-alanine; Shibatani T et al.; The formation of L-asparate beta-decarboxylase by Pseudomonas dacunhae was compared on media containing a variety of organic acids and amino acids as a carbon source . Although the enzyme was formed constitutively when the organism was grown on basal medium or on that containing tricarboxylic acid cycle intermediates, it was induced twofold by L-glutamate and repressed one-tenth by L-serine . L-Glutamine, L-proline, L-leucine, glycine, and L-threonine also showed induction effects lower than that of L-glutamate . L-Glutamate derepressed the serine effect . This glutamate effect was observed effect was observed with other microoganisms, e.g., Achromobacter pestifer and Achromobacter liquidum . Since the intermediates from L-glutamate metabolism had no effect, this induction effect was specific to L-glutamate . The formation of some glutamate-related enzymes was measured and is discussed in relation to the formation of L-asparate beta-decarboxylase . L-Asparate beta-decarboxylase was purified to an electrophoretically homogenous state from L-glutamate-grown cells of P . dacunhae, and some properties were compared with those of the enzyme from fumarate-grown cells . The two enzymes were identical in disc electrophoresis, molecular weight, and some enzymatic properties . The industrial production of L-alanine from L-aspartic acid acid was improved by using the culture broth with highly induced L-asparate beta-decarboxylase (9.4 U/ml of broth).

Prikl Biokhim Mikrobiol, 1979 Sep-Oct, 15(5), 790 - 2
{Products of bacterial destruction of sodium dodecyl sulphate}; Stavskaia SS et al.; The products of Na dodecyl sulphate destruction by the three bacterial cultures--Pseudomonas aeruginosa, Flavobacterium devorans, Achromobacter guttatus--were examined . The cultures were shown to decompose Na dodecyl sulphate in a similar way . The primary mechanism of destruction was found to be hydrolysis of the sulpho-ester bond in the molecule, leading to the separation of sulphate-ion and formation of dodecanol . Products of bacterial destruction of alkyl sulphates did not show foam forming capacity.

Mikrobiologiia, 1979 Jul-Aug, 48(4), 604 - 9
{Effect of formate on the respiration of different microorganisms}; Zakharova EV; The cells of methylotrophic bacteria (Achromobacter parvulus 1, Pseudomonas methylica 2 and 20, Ps . fluorescens 45, Ps . oleovorans 52) and Candida methylica 101 grown in a medium with methanol take up oxygen in the presence of formiate . A . parvulus is most resistant to formiate (KM = 2.3 +/- 0.5) X 10(-3) M; Ki = 4 x 10(-1) M) . When grown on Cn substrates, these microorganisms cannot oxidize formiate (with an exception of A . parvulus) . Formiate also serves as a respiration substrate for E . coli K-12 and the purple bacterium Rhodopseudomonas palustris, disregarding the composition of a medium on which the cells are grown . When the cells cannot oxidize formiate, it inhibits the respiration of the cells and cell extracts in the presence of other substrates, including TMPD + ascorbate . The respiration is completely inhibited at a concentration of formiate of 10(-3)--10(-2) M . It follows therefore that formiate acts on the terminal cytochrome oxidase, regardless of whether it is cytochrome a or o.

Cancer Treat Rep, 1979 Jun, 63(6), 1127 - 32
Treatment of L5178Y tumor-bearing BDF1 mice with a nonimmunogenic L-glutaminase-L-asparaginase; Abuchowski A et al.; An L-glutaminase-L-asparaginase from Achromobacter has been rendered nonimmunogenic by the covalent attachment of polyethylene glycol (PEG) to nonessential amine groups of the enzyme . PEG-L-glutaminase-L-asparaginase exhibits a greatly enhanced half-life in the bloodstream compared to the unmodified enzyme in normal mice, and is effective in prolonging the survival of BDF1 mice inoculated ip with L5178Y cells . PEG-L-glutaminase-L-asparaginase appears rapidly in the blood following ip injection.

Biochem J, 1979 Apr 1, 179(1), 53 - 8
Differences in the degradation of native collagen by two microbial collagenases; Lecroisey A et al.; The early stages of degradation of native collagen by two bacterial collagenases were studied by electron microscopy and by automatic Edman degradation . The purified collagenase from Clostridium histolyticum was shown to cleave native collagen at several sites, but not progressively from the N-terminus, as had been previously suggested . The homogeneous collagenase from Achromobacter iophagus cleaves native collagen preferentially at two sites corresponding to the interbands 33-34 and 41-42 . The latter lies within the region cleaved by the eukaryotic collagenases.

J Clin Microbiol, 1979 Mar, 9(3), 425 - 36
Achromobacter species (CDC group Vd): morphological and biochemical characterization; Chester B et al.; Twenty-three isolates of Achromobacter species (CDC group Vd) were examined morphologically and biochemically . Gram stains revealed gram-variable bacilli frequently curved or hooked at one pole and often coryneform in shape and arrangement . Electron microscopy revealed the presence of extracellular material in polar accumulations and demonstrated the polar flagella arrangement seen by light microscopy to be lateral . Two colony types were produced; one was minute and watery at 24 h (35 degrees C) progressing to large, mucoid colonies at 48 h, and the other type was shiny, glistening, opaque but nonmucoid . All isolates grew on MacConkey agar and produced catalase, oxidase, and urease . Most grew on salmonella-shigella agar, reduced nitrate to nitrite and gas, hydrolyzed esculin, deaminated phenylalanine (2 to 4 days) and produced H2S in triple sugar iron agar (4 to 12 days) . Oxidation of carbohydrates was weak, delayed, and limited to glucose and xylose . Two isolates also oxidized maltose, mannitol, and sucrose . The ability of miniaturized "nonfermenter" kits to identify Achromobacter species was tested . The Minitek (Baltimore Biological Laboratory, Cockeysville, Md.) and N/F (Corning, Roslyn, N.Y.) systems, respectively, identified 21 and 19 of the 23 isolates, whereas the Oxi/Ferm (Roche, Nutley, N.J.) identified 13 and the API 20E (Analytab Products, Plainview, N.Y.) identified only 3.

Mol Cell Biochem, 1979 Jan 26, 23(2), 87 - 108
Some newly characterized collagenases from procaryotes and lower eucaryotes; Keil B; Chemical and enzymatic properties of four collagenases newly isolated from anaerobic Clostridium histolyticum, aerobic Achromobacter iophagus, and from two lower eucaryotes, the fungus Entomophthora coronata and the insect Hypoderma lineatum are reviewed . The problems of their biosynthesis and precursors, namely the effect of induction of collagenase and neutral proteinase in Achromobacter by their macromolecular substrates are discussed . The two bacterial collagenases are Zn-metallo-enzymes; the highly purified Clostridium collagenase contains cyst(e)ine, serine phosphate and tryptophan additionally to amino acids reported previously . Achromobacter collagenase has the highest specific activity of all collagenases; it yields by autolysis enzymatically active degraded forms . The active dimer is composed of two identical subunits of molecular weight 35,000 . Similarities between Achromobacter collagenase, thermolysin and Bacillus subtilis neutral proteinase in molecular weight, amino acid composition, and amino acids important for the active sites are discussed . The two collagenases from low eucaryotes are serine proteinases; Hypoderma collagenase is homologous to the trypsin family in the amino terminal sequence . The initial cleavage of native collagen by highly purified bacterial collagenases occurs in the central helical part of the alpha chains and not progressively from the amino terminal end . One of the two initial cleavages produced by Achromobacter collagenase is situated in the region cleaved specifically by vertebrate collagenases, but with different bond specificity . The same is true for the insect collagenase . Entomophthora collagenase is a proteinase of broad specificity which also cleaves collagen in its helical parts . All four collagenases also degrade other proteins according to their bond specificity.

Zentralbl Bakteriol Naturwiss, 1979, 134(6), 507 - 12
Studies on the aerobic non-symbiotic nitrogen-fixing bacteria, other than Azotobacter, in Egyptian soils; Abd-el-Malek Y et al.; Twenty isolates of micro-organisms capable of growing on nitrogen-deficient medium and found as contaminants in Azotobacter cultures were isolated from Egyptian soils and studied for their morphological, cultural, and physiological properties . These micro-organisms s are members of Rhizobiaceae, Pseudomonadaceae, Achromobacteriaceae, Enterobacteriaceae, Micrococcaceae, Bacillaceae, and Streptomycetaceae as well as some yeasts . In nitrogen-free medium the micro-organism fixed only small amounts of atmospheric nitrogen, hardly exceeding 3 ppm and because of their low sugar consumption rates, efficiences of N2-fixation sometimes reaching 18 mg nitrogen fixed/g carbon oxidized were recorded . Addition of 15 ppm combined nitrogen to the medium increased the amounts of nitrogen fixed to 3--9 ppm.

J Gen Virol, 1978 Nov, 41(2), 265 - 72
Phage growth characteristics on stationary phase Achromobacter cells; Robb SM et al.; The growth characteristics of alpha3a bacteriophage on stationary phase Achromobacter strain 14 are described . Phage alpha3a growth on stationary phase cells is characterized by a long and variable latent period of 6 to 9 h and an increased burst size of 710 p.f.u./cell as compared with 153 p.f.u./cell in exponential wild type cells . During the latent period the infected cells are very sensitive to changes in growth conditions and in particular, dilution . Pre-conditioning of the bacterial cells by allowing them to stand for 24 h after shaking for 3 days is an important aspect of the stationary phase phage growth system . Cells which have been allowed to stand retain the ability to be infected and to support phage growth for at least 16 days . Shaking cultures gradually lose the ability to support phage growth but the phage can persist in the host cell for 10 days until removal from shaking when the lytic cycle can proceed after allowing the cultures to stand.

Am J Clin Pathol, 1978 Oct, 70(4), 712 - 7
Community-acquired bacteremic Achromobacter xylosoxidans type IIIa pneumonia in a patient with idiopathic IgM deficiency; Dworzack DL et al.; A man was hospitalized with bacteremic Achromobacter xylosoxidans type IIIa pneumonia . The authors are aware of no previously reported similar infections caused by this bacterium . A clinical cure was achieved with a combination of carbenicillin and kanamycin therapy . Microtiter susceptibility testing revealed that carbenicillin was the antibiotic to which A . xylosoxidans IIIa was most sensitive (minimal inhibitory concentration, 1.6 microgram/ml) and that synergy between carbenicillin and kanamycin existed . During the patient's hospitalization, deficiency of IgM (21 mg/dl) was found . Specific serum activity against A . xylosoxidans IIIa was detected by the agglutination method . Specific anti-A . xylosoxidans IIIa IgG, but not IgM, was detected by indirect immunofluorescence . It appears that a defect in immunologic recognition of A . xylosoxidans IIIa as an invasive bacterium, a defect in synthesis of specific IgM, or both, contributed to this patient's infection.

J Clin Microbiol, 1978 Jul, 8(1), 61 - 6
Identification of Achromobacter species by cellular fatty acids and by production of keto acids; Dees SB et al.; The cellular fatty acid composition and metabolic products of 12 reference strains of Achromobacter sp . and A . xylosoxidans were determined by gas-liquid chromatography (GLC) . Results showed that the two Achromobacter groups are strikingly different and can be readily distinguished on the basis of cellular fatty acids and the short-chain acids produced by Achromobacter sp . The major cellular fatty acids of Achromobacter sp . were octadecenoic (18:1) and a 19-carbon cyclopropanoic (19:0 delta) acid, whereas hexadecanoic (16:0) and a 17-carbon cyclopropanoic (17:0 delta) acid were principal components of the lipids of A . xylosoxidans . Hydroxy acids were not found in strains of Achromobacter sp . but comprised approximately 20% of the cellular fatty acids of A . xylosoxidans . In addition, Achromobacter sp . produced relatively large amounts of 2-ketoisocaproic acid, which was detected in only trace amounts from strains of A . xylosoxidans . The data show that GLC tests provide additional criteria for differentiating groups which are very closely related when evaluated with conventional tests . The GLC tests can be readily adapted in the clinical laboratory because they are rapid, highly reproducible, relatively inexpensive, and simple to perform.

Experientia, 1978 May 15, 34(5), 579 - 80
Bacterial utilization of cyclo(glycl-L-prolyl); Adams E et al.; 3 strains of soil bacteria (2 Achromobacter, 1 Flavobacterium) were isolated by growth on cyclo(Gly-L-Pro) as carbon/nitrogen source . Good growth required yeast extract supplements . Utilization of cyclo(Gly-L-Pro) was inducible . Many efforts failed to obtain active cell-free preparations . Injected radioactive cyclo(Gly-L-Pro) was excreted intact by the albino rat; in contrast, injected radioactive H-Gly-L-Pro-OH was extensively metabolized.

Med Klin, 1978 Mar 24, 73(12), 423 - 7
{In vitro activity of cefoxitin compared with the activity of other antibiotics and trimethoprim-sulfamethoxazole (author's transl)}; Freitag V; The new developed antibiotic cefoxitin has been tested in vitro on its effectiveness against bacteria isolated from human material . Pseudomonas aeruginosa and streptococci of the serological group D were not tested, because these microorganisms are not sensitive against this antibiotic . Besides cefoxitin other antibiotics (cephalothin, cephalexin, ampicillin, carbenicillin, tetracycline, gentamicin, penicillin G, oxacillin) and trimethoprim-sulfamethoxazole were also tested . As to the results cefoxitin shows the best effectiveness against all gram-positive bacteria and against gram-negative bacteria including indol-positive species of proteus, providencia, serratia marcescens and the anaerobe, gram-negative, non-spore-forming rods . There are different results with enterobacter-, citrobacter-, achromobacter- and acinetobacter-species.

Biochim Biophys Acta, 1978 Mar 14, 523(1), 75 - 81
Properties of taurine: alpha-ketoglutarate aminotransferase of Achromobacter superficialis . Inactivation and reactivation of enzyme; Toyama S et al.; The activity of taurine: alpha-ketoglutarate aminotransferase (taurine: 2-oxoglutarate aminotransferase, EC 2.6.1.55) from Achromobacter superficialis is significantly diminished by treatment of the enzyme with (NH4)2SO4 in the course of purification, and recovered by incubation with pyridoxal phosphate at high temperatures such as 60 degrees C . The inactive form of enzyme absorbing at 280 and 345 nm contains 3 mol of pyridoxal phosphate per mol . The activated enzyme contains additional 1 mol of pyridoxal phosphate with a maximum at 430 nm . This peak is shifted to about 400 nm as a shoulder by dialysis of the enzyme, but the activity is not influenced . The inactive form is regarded as a partially resolved form, i.e . a semiapoenzyme . The enzyme catalyzes transamination of various omega-amino aicds with alpha-ketoglutarate, which is the exclusive amino acceptor . Hypotaurine, DL-beta-aminoisobutyrate, beta-alanine and taurine are the preferred amino donors . The apparent Michaelis constants are as follows; taurine 12 mM, hypotaurine 16 mM, DL-beta-aminoisobutyrate 11 mM, beta-alanine 17 mM, alpha ketoglutarate 11 mM and pyridoxal phosphate 5 micron.

Mikrobiologiia, 1978 Mar-Apr, 47(2), 338 - 41
{Microorganisms that break down alkyl sulfates}; Rotmistrov MN et al.; The following bacteria assimilating alkyl sulphates as a sole source of carbon and energy were isolated from various substrates: Pseudomonas, Achromobacter, Flavobacterium, Citrobacter, Enterobacter . The bacteria decomposed sodium dodecyl sulphate at a high rate, and some of them, industrial preparations of alkyl sulphates . The ability to assimilate alkyl sulphates was tested in 257 collection cultures belonging to different taxonomic groups . Alkyl sulphates were found to be decomposed by heterotrophous gram-negative rod-like bacteria belonging to different families and genera . The frequency of bacteria destroying alkyl sulphates at a high rate was found most often in the Pseudomonas genus.

J Clin Pathol, 1978 Feb, 31(2), 156 - 61
Cerebral ventriculitis associated with Achromobacter xylosoxidans; Shigeta S et al.; Six patients in the neurosurgical ward of Fukushima Medical College Hospital suffering from ventriculitis due to Achromobacter xylosoxidans infection had undergone craniotomy or cranial trepanation before the infection . The strains of A . xylosoxidans isolated from the patients were resistant to streptomycin, ampicillin, cephaloridine, gentamicin, and colistin . They were also resistant to chlorhexidine digluconate (Hibitane) in a concentration of 2% . When a study of the chlorhexidine used in the hospital was carried out four strains of A . xylosoxidans were isolated from 20 containers of chlorhexidine solution in the surgical ward but not from those in the operating theatre.

J Clin Microbiol, 1978 Feb, 7(2), 239 - 41
Achromobacter xylosoxidans isolates in Hawaii; Pien FD et al.; Clinical and bacteriological features of nine cases in which Achromobacter xylosoxidans were isolated in Hawaii are described . Five cases were ear infections mixed with other gram-negative bacteria . Colonial morphology, xylose oxidation, peritrichous flagella staining, and antibiotic susceptibility pattern assisted in separating this bacterium from other nonfermentative, oxidase-positive, gram-negative rods.

Biotechnol Bioeng, 1978 Feb, 20(2), 255 - 66
Continuous production of NADP by immobilized Achromobacter aceris cells; Uchida T et al.; Several microorganisms having higher nicotinamide adenine dinucleotide kinase (NAD kinase, EC 2.7.1.23) activity were immobilized into polyacrylamide gel lattices . The enzyme activity field by immobilization was highest in Achromobacter aceris AKU 0120 . By the incubation of the immobilized A . aceris cells at pH 4.0, the NAD kinase activity increased and the adenosine triphosphate (ATP)-degradation activity disappeared completely . Enzymatic properties of the immobilized A . aceris cells were investigated and compared with those of intact cells . The optimal pH and the optimal temperature of immobilized cells were the same as those of intact cells . Immobilized cell NAD kinase was more stable than that of intact cells . The operational half-life of immobilized cells was 20 days when the substrate solution was passed through a column packed with immobilized cells at a flow rate which gives a space velocity (SV) of 0.1 hr-1 at 37 degrees C . On the other hand, the half-life of the intact cells was only 6 hr.

Biochim Biophys Acta, 1978 Jan 12, 522(1), 218 - 28
Subunit structure of Achromobacter collagenase; Keil-Dlouha V et al.; The highly active form of collagenase (EC 3.4.24.3) from Achromobacter iophagus (specific activity 2 microkat/mg) has a molecular weight of 70,000 and the sedimentation coefficient s20,2 = 4.4 S . It is composed of two subunits of molecular weight 35,000 and s20,w of 2.9 S . The dissociation of the dimer under different conditions resulted in the complete and irreversible loss of enzymic activity . A unique N-terminal sequence Thr-Ala-Ala-Asp-Leu-Glu-Ala-Leu-Val- indicates that the two subunits are identical, at least in the N-terminal part of the polypeptide chain . Reduction and pyridylethylation of the subunit change neither molecular weight nor amino acid composition: therefore each subunit of molecular weight 35,000 consists of a single polypeptide chain . Another active and homogeneous form of Achromobacter collagenase (specific activity 1.64 microkat/mg) gives a value for the apparent molecular weight of 80,000 on sodium dodecyl sulphate-polyacrylamide electrophoresis . It is also a dimer in which each of the two subunits of molecular weight 35,000 binds non-covalently a peptide of molecular weight 5000 . The dissociation of this form of collagenase is also accompanied by irreversible loss of enzymic activity . The amino acid composition of the subunits which were isolated from both 70,000 and 80,000 collagenases is the same . The role of dimer-monometer equilibrium in the biological function of collagenase is discussed.

Aust Vet J, 1977 Nov, 53(11), 542 - 4
Observation on the bacterial population of the os cervix of the ewe before and after embryo death; Sawyer GJ; Bacterial samples were isolated from the os cervix of mature Merino ewes during a normal oestrous cycle and after colchicine treatment which killed the 25-day old embryos . The population of bacteria was small during the oestrous cycle and consisted of Achromobacter spp . Alcaligenes spp, Corynebacterium spp, Bacillus spp and Escherichia coli . There was a significant increase in the numbers of bacteria isolated and a change in the proportions of the bacteria isolated from ewes after embryo death . Much greater numbers of potentially pathogenic organisms, including Pasteurella multocida, Corynebacterium pyogenes, Staphylococcus spp and Streptococcus spp were isolated after embryos were killed . It is suggested that persistence of potential pathogens in the vagina may impair fertility in ewes at the first oestrus after embryo death.

Appl Environ Microbiol, 1977 Oct, 34(4), 351 - 4
Metabolism of quaternary carbon compounds: 2,2-dimethylheptane and tertbutylbenzene; Catelani D et al.; Two Achromobacter strains capable of utilizing 2,2-dimethylheptane or tertbutylbenzene as the sole carbon and energy source were isolated from waste-water . Pivalic acid was found in the cultures of Achromobacter A1 containing 2,2-dimethylheptane . From cultures of Achromobacter A2 in the presence of tertbutylbenzene, a diol was isolated and identified as 2,3-dihydro-2,3-dihydroxytertbutylbenzene . Evidence for meta cleavage of the aromatic ring and for accumulation of pivalic acid in the cultures was also obtained . A metabolic pathway for tertbutylbenzene is suggested.

Mol Cell Biochem, 1977 Sep 9, 17(2), 61 - 73
Microassay with the NADH-induced light reaction, technique improved by means of purified enzymes from Achromobacter fischeri; Brolin SE et al.; Purification of a commercial preparation of Achromobacter fischeri was carried out by agarose-suspension electrophoresis and by molecular-sieve chromatography . Both the luciferase and an oxidoreductase, catalyzing reduction of FMN with NADH, were obtained in more than one form . Flavins, liable to interfere with the light production in analytical applications, were present in amounts worthy of consideration, but seem to be firmly bound to protein . The major quantity was found in the enzymatically inactive fractions . In free zone electrophoresis of the main luciferase component, the mobility of the zone containing enzyme activity was calculated to -4.0 X 10(-5) cm2 sec-1 V-1 at 12 degrees C . Fractions of the two enzymes were separated and mixed in different proportions to study how the intensity and time course of NADH-induced light emission can be modified . These experiments disclosed how reaction mixtures will have to be composed in appropriate photokinetic assays, using NADH as measurable product . A regenerating system based on the purified fractions is described . Instead of the light flash, following the consumption of NADH, the light is emitted on a well maintained level, permitting assays with a less elaborate equipment than the one required for the recording of fast reactions.

Mikrobiologiia, 1977 Sep-Oct, 46(5), 885 - 9
{Pathways of assimilation of carbon oxides in carboxydobacteria Seliberia carboxydohydrogena and Achromobacter carboxydus}; Romanova AK et al.; Assimilation products of 14C-bicarbonate and carbon-14C oxide were studied in two carboxydobacteria Seliberia carboxydohydrogena and Achromobacter carboxydus which differed in their ability for chemolithoautrophous growth in the presence of hydrogen . The dynamics and composition of labeled products formed upon assimilation of 14C-bicarbonate in the presence of unlabeled carbon oxide by the two organisms, the composition of products formed upon assimilation of 14CO by suspensions of S . carboxydohydrogena Z-1062 during 5 minutes, and the dynamics and composition of labeled assimilates of A . carboxydus Z-1171 after incubation in the presence of 14CO, were found to be consistent with those expected in the action of the reductive pentose phosphate Calvin cycle . The similarity of products formed upon assimilation of 14CO2 and 14CO suggests that CO is first oxidized to CO2, and only is assimilated.

J Biochem (Tokyo), 1977 Aug, 82(2), 535 - 43
L-Lysine-alpha-ketoglutarate epsilon-aminotransferease . Properties of the bound pyridoxal 5'-phosphate; Misono H et al.; L-Lysine-alpha-ketoglutarate epsilon-aminotransferase of Flavobacterium lutescens (Achromobacter liquidum) IFO 3084 shows positive circular dichroic bands at 340 and 415 nm where absorption maxima are observed, and fluorescence maxima at 380 and 490 nm on excitation at 340 and 415 nm, respectively . The pyridoxal 5'-phosphate absorbing at 415 nm is bound through an aldimine linkage to an epsilon-amino group of the lysine residue of the protein . Upon aging, the 415 nm pyridoxal 5'-phosphate changes to a less active form (lambda max, 325 nm), which is distinguishable from the 340 nm pyridoxal 5'-phosphate . This 325 nm bound pyridoxal 5'-phosphate is reduced with sodium borohydride and shows no circular dichroism . When the semiapoenzyme is aged under the same conditions, no spectral change is observed . These findings suggest that the pyridoxal 5'-phosphate bound through an aldimine linkage may be converted into a carbinol amine or some other related form by aging.

J Clin Pathol, 1977 Jul, 30(7), 595 - 601
Strains of Achromobacter xylosoxidans from clinical material; Holmes B et al.; Eleven strains of Achromobacter xylosoxidans have been received from among 1106 strains of Gram-negative, non-fermentative bacteria submitted to the National Collection of Type Cultures for computer-assisted identification since 1 January 1972 . The strains showed resistance to a wide range of antimicrobial agents and five of the isolates possibly played a pathogenic role . The biochemical characteristics of these 11 strains were compared with those of three culture collection strains.

Mikrobiologiia, 1977 May-Jun, 46(3), 570 - 7
{Species composition of heterotrophic bacteria in the water of the Rybinek reservoir}; Lapteva NA; Twenty four cultures of heterotrophic bacteria were isolated from the water of the Rybinsk Reservoir on a medium containing organic matter . Bacteria belonging to the families Pseudomonadaceae and Achromobacteriaceae were most widely distributed; bacteria of the Mycobacteriaceae family were encountered less often . Predominating species which belonged to the genera Pseudomonas, Caulobacter, Flavobacterium, Mycobacterium, and Corynebacterium were isolated from the central part of the Reservoir . The special composition depended on the season . Bacteria of the Caulobacter genus were isolated mainly by the end of July and in May . Bacteria of the Corynebacterium genus predominated in March . Microflora belonging to the genera Pseudomonas and Flavobacterium was more constant . Bacteria isolated on media with the minimum content of organic matter (0.5 mg C per litre) belonged mainly to the Pseudomonas genus . Morphological, cultural, and physiological properties of the bacteria were studied . The bacteria grew on media containing carbohydrates and alcohols as a source of carbon though organic acids were assimilated to a less extent . The activity of catalase was found in the cultures, with an exception of the genera Caulobacter and Achromobacter.

J Gen Virol, 1977 Apr, 35(1), 117 - 23
Rifampicin-resistant mutant supporting bacteriophage growth on stationary phase Achromobacter cells; Robb SM et al.; A rifampicin-resistant Achromobacter mutant with an altered RNA polymerase was isolated . The mutant supports phage alpha3a growth in both log and stationary phase cells . Phage growth on stationary phase cells is sensitive to aeration and growth only occurs at oxygen concentrations of less than 5-2 p.p.m . The rifampicin-resistant mutant is similar to the spontaneous mutant strain 14 reported by Woods (1976) in that both mutants support stationary-phase phage growth under micro-aerophilic conditions . The isolation of the rifampicin-resistant mutant with an altered RNA polymerase suggests that the phenomenon of stationary phase phage growth could be due to a change in the template specificity of the Achromobacter RNA polymerase . Plaque morphology mutants which grow on log and/or stationary phase cells of the Achromobacter wild type, strain 14 and rifampicin-resistant strains are also described.

J Biol Chem, 1977 Mar 25, 252(6), 2039 - 45
Proteolytic activation of rat liver adenylate cyclase by a contaminant of crude collagenase from Clostridium histolyticum; Hanoune J et al.; Treatment of rat liver plasma membranes with various commercial preparations of crude collagenase from Clostridium histolyticum at concentrations as low as 1 mug/ml, resulted in activation of the adenylate cyclase system . Maximal activation occurred at 50 to 100 mug/ml of collagenase, and promoted a 2- to 3-fold increase in the basal activity as well as in the activities stimulated by catecholamines, glucagon, fluoride, or GTP . This was due to an increase in the maximal velocity of the cyclizing reaction without any increase in the affinity of the enzyme for its substrate . Treatment of plasma membranes with crude collagenase did not induce gross structural modifications as judged by electron microscopic examination . 5'-Nucleotidase activity was slightly inhibited and ATPase activity remained unaffected . The stimulatory substance was nondialyzable, thermolabile, and inhibited by both EDTA and -SH reagents, thus appearing to be a protein . The following observations suggest the effects observed were due to other protease(s) present in crude collagenase: (a) only crude collagenase was active on liver adenylate cyclase: treatment with purified collagenase from C . histolyticum or from Achromobacter iophagus gave no stimulation; (b) the stimulatory activity was irreversible since washing of the membranes after treatment was without effect; (c) crude collagenase contained no lecithinase or sphingomyelinase activity under our conditions of adenylate cyclase assay; (d) after chromatography on Sephadex G-100, the activator appeared as a peak in the 30,000-dalton region and was clearly separated from the collagenase and clostripain peaks, but coincident with elastolytic and caseinolytic activities; (e) the effect of crude collagenase could be prevented by addition of elastin in vitro and was mimicked by purified elastase from hog pancreas . It remains to be seen whether the effects observed result from an increase in the catalytic constant of adenylate cyclase, or an unmasking of new catalytic sites.

Mikrobiologiia, 1977 Mar-Apr, 46(2), 368 - 70
{Content of nitrogen bases in the DNA of carboxide bacteria}; Savel'eva ND; The composition of nitrogen bases was determined in total DNA of live strains of carboxide bacteria belonging to the genera Pseudomonas, Comamonas, Seliberia (Agrobacterium), Achromobacter . Considerate differences in the nucleotide composition of DNA of the studied bacteria confirm that they indeed belong to different genera . At the same time, the content of GC pairs in DNA of these bacteria is within the range found for the corresponding genera by other authors.

Biotechnol Bioeng, 1977 Mar, 19(3), 301 - 9
Bacteriolysis by immobilized enzymes; Karube I et al.; Bacteriolytic enzymes produced by Achromobacter lunatus were immobilized in collagen membrane . Intact bacteria such as Pseudomonas solanacearum, Xanthomonas oryzae, Staphylococcus aureus, and Pseudomonas aeruginosa were lyzed with the bacteriolytic enzyme-collagen membrane . Relative activity of the bacteriolytic enzyme-collagen membrane against Pseu . solanacearum was about 2% of that of native bacteriolytic enzymes . No difference in the optimum pH was observed between immobilized enzymes and native enzymes . The bacteriolytic enzymes in the collagen membrane were stable against sodium chloride which was an inhibitor of the native bacteriolytic enzymes . Xanthomonas oryzae and Pseu . aeruginosa were continuously lyzed by a reactor containing the rolled bacteriolytic enzyme-collagen membrane.

Z Allg Mikrobiol, 1977, 17(2), 139 - 48
{Physiology of aniline catabolism by achromobacter Ir2}; Rabsch W et al.; A bacterium was isolated from soil which utilizes aniline as sole source of carbon and nitrogen . It was identified as Achromobacter sp . The cells grow at concentrations in the range of 0.5 to 1.25 mg aniline/ml with a growth rate of 0.3 h-1 . Substrate inhibition was observed at concentrations higher than 1.5 mg/ml, 3.0 mg/ml completely inhibit the growth . The yield coefficient was 0.63 . Aniline was degraded with an activity of 200 microng/mg cell dry weight/hour . Aniline was assimilated and completely degraded . The remaining nitrogen was quantitatively detected as ammonia . The enzyme system involved in aniline degradation was induced by aniline but not repressed by succinate and ammonia.

Zentralbl Bakteriol {Orig A}, 1976 Dec, 236(4), 513 - 30
{Culture and differentiation of obligatory aerobic gram-negative rods from human material; a scheme for application in routine diagnosis (author's transl)}; von Graevenitz A et al.; The diagnosis of obligately aerobic Gram-negative rods in the clinical laboratory may encounter difficulties since media used for Enterobacteriacae are only partially usable for the diagnosis of this group of bacteria (Psuedomonas, Xanthomonas, Alcaligenes, Achromobacter, Brucella, Bordetella, Flavobacterium, Moraxella, Acinetobacter, and some still unnamed taxa) . We have developed a diagnostic scheme, based on recent publications in the field and representing an extension of earlier tables from this and other laboratories, which attempts to classify a maximal number of obligately aerobic Gram-negative rods with a minimal number of tests . The scheme, employed on 4051 strains, used blood agar and MacConkey Agar as isolation media . Growth characteristics on these media and microscopic morphology may be of help, but only the type of growth on Triple Sugar Iron (or Kligler's) Agar is characteristic for the group as a whole (no growth in the butt, alkalinization or no pH change on the slant) . A primary identification series employs tests for oxidase (Kovacs), oxidation of glucose and xylose (in OF medium), deoxyribonuclease and indole (in DNase Test Agar with Methyl Green), nitrate reduction (in Indole Nitrite Medium), motility (hanging drop), and fluorescein production (on Flo Agar) . Results of Kirby-Bauer antimicrobial sensitivity testing serve as additional (colistin) or confirmatory criteria . Incubation is at 30 degrees C for 24-48 hrs . If a diagnosis is not possible than, a secondary series, including tests for lysine decarboxylase (tablets), 4 hr urease, esculin hydrolysis, growth at 42 C and on SS Agar, gelatin liquefaction, and flagellar staining may have to be used, and read after 4-24 hrs at 30 degrees C . Five tables, drawn up according to oxidase, glucose, and xylose reactions, serve to identify the species or taxa . Biotypes cannot be differentiated . The scheme will need updating as more knowledge of these bacteria will become available.

Mikrobiologiia, 1976 Nov-Dec, 45(6), 973 - 8
{Effect of nitrogen sources on growth and cholesterol decomposing activity of Mycobacterium rubrum and Achromobacter canadicans}; Imshenetskii AA et al.; The effect of 12 sources of nitrogen on growth and cholesterine-decomposing activity was studied with Mycobacterium rubrum and Achromobacter candicans . The yield of biomass and the rate of cholesterine decomposition depended on the source of nitrogen and its concentration in the medium . The highest specific activity of the enzyme decomposing cholesterine was found during growth of the cultures on media containing reduced forms of nitrogen . The activity of the enzyme of Mycobacterium rubrum was by 25% higher than that of Achromobacter candicans . The optimum conditions for the production of the enzyme by Mycobacterium rubrum were on media containing 1.0 g of asparagine or 5.0 g of ammonium nitrate per one litre, and for the enzyme production by Achromobacter candicans, on media containing 5.0 g of ammonium phosphate per one litre.

Appl Environ Microbiol, 1976 Nov, 32(5), 694 - 8
Bacteria within ovules and seeds; Mundt JO et al.; Surface-sterilized ovules and seeds of 27 species of plants were cultured in the water of syneresis of a nutrient medium low in agar content . Bacteria were obtained from 30% of the ovules, 15% of the seeds of herbaceous plants, 16% of the seeds of woody plants, 5.4% of the overwintered noncereal seeds, and 13.5% of overwintered cereal seeds . In no instance did every ovule or seed of a plant species contain bacteria . No bacteria were obtained from the hard, waxy seeds of mimosa or yellowwood . They were not obtained from ovules with unbroken coats or from seeds with coats that were not ruptured during the swelling of the seed . Only one species of bacteria was recovered in 93% of the instances in which bacteria were obtained . Bacteria were obtained from seeds that were embedded in the acidic parenchyma of the lemon or surrounded by the thickened flesh of the cucurbits . The bacteria were distributed among 19 genera and 46 species . The species isolated in greatest numbers were Bacillus megaterium, B . cereus, Erwinia herbicola, Flavobacterium devorans, and Pseudomonas fluorescens . Bacteria recovered less frequently were in the genera Achromobacter, Acinetobacter, Alcaligenes, Brevibacterium, Corynebacterium, Cytophaga, Leuconostoc, Micrococcus, Nocardia, Proteus, Streptococcus, Streptomyces, and Xanthomonas . Members of 11 genera and 15 species of bacteria were isolated once.

Mikrobiologiia, 1976 Sep-Oct, 45(5), 911 - 4
{Survival of epiphytic microorganisms following sublimation dehydration and storage}; Shushunova AV et al.; Epiphytic microorganisms belonging to the genera Pseudomonas, Agrobacterium, Achromobacter, Flavobacterium, Mycobacterium, Mycococcus, Nocardia, Corynebacterium, Arthrobacter, Bacillus, and Microbacterium were dehydrated by sublimation employing two suspension media, and were stored at 3-5 degrees C during 1-2.5 years . Survival of epiphytic microorganisms upon dehydration by sublimation and storage was found to depend on their taxonomy (genus, species, strain) and the composition of the suspension medium . The cultures can be stored after dehydration during 2-2.5 years.

J Med Microbiol, 1976 Aug, 9(3), 345 - 53
Acinetobacter and similar organisms in ear infections; Dadswell JV; Fifty-seven strains of acinetobacter-like organisms were isolated over a period of 26 months from the ears of 55 patients with acute or chronic otitis media, or otitis externa, and one strain was isolated in a survey of 50 normal ears . After comparison with eight reference strains, 32 of the isolates were identified as Acinetobacter anitratus, 22 as Acinetobacter Iwoffii, three as Moraxella spp . and one as Achromobacter sp . Analysis of the clinical findings suggests that although most of these organisms played little part in the disease process, a few strains were probably pathogenic in this situation.

Appl Environ Microbiol, 1976 Aug, 32(2), 250 - 6
Effects of thermoradiation on bacteria; Pallas JE 3rd et al.; A 60Co source was used to determine the effects of thermoradiation on Achromobacter aquamarinus, Staphylococcus aureus, and vegetative and spore cells of Bacillus subtilis var . globigii . The rate of inactivation of these cultures, except vegetative-cell populations of B . subtilis, was exponential and in direct proportion to temperature . The D10 (dose that inactivates 90% of the microbial population) value for A . aquamarinus was 8.0 Krad at 25 degrees C and 4.9 Krad at 35 degrees C . For S . aureus, D10 was 9.8 and 5.3 Krad at 35 and 45 degrees C, respectively . Vegetative cells of B . subtilis demonstrated a rapid initial inactivation followed by a steady but decreased exponential rate . The D10 at 25 degrees C was 10.3 Krad, but at 35 and 45 degrees C this value was 6.2 and 3.8 Krad, respectively . Between 0 and 95 Krad, survival curves for B . subtilis spores at 75 degrees C showed slight inactivation, increasing in rat at and above 85 degrees C . The D10 values for spores at 85 and 90 degrees C were 129 and 92 Krad, respectively . Significant synergism between heat and irradiation was noted at 35 degrees C for A . aquamarinus and 45 degrees C for S . aureus . The presence of 0.1 mM cysteine in suspending media afforded protection to both cultures at these critical temperatures . On the other hand, cysteine sensitized B . subtilis spores at radiation doses greater than 100 Krad . The combined effect of heat and irradiation was more destructive to bacteria than either method alone.

J Gen Virol, 1976 Jul, 32(1), 45 - 50
Bacteriophage growth on stationary phase Achromobacter cells; Woods DR; A new phage-host system is described in which phage alpha 3a grows on stationary phase Achromobacter mutant strains . Characteristic clear plaques are formed, at an e.o.p . 10(-1) to 10(-2), on already confluent bacterial lawns of the mutant strains . Phage growth is sensitive to aeration and growth only occurs under micro-aerophilic conditions . Lysates prepared on the mutant strains cannot transduce in contrast to transducing lysates prepared from wild type Achromobacter strains.

J Gen Microbiol, 1976 Jun, 94(2), 351 - 8
Divalent cations in the envelope of a psychrophilic Achromobacter; Ledebo I; The function of Ca2+ in a psychrophilic Achromobacter, previously found to bind large amounts of these ions to its envelope, has been studied . Bacteria suspended in media of low ionic content showed decreases in wet weight, dry weight and growth capacity, and increases in light scattering and in the release of u.v.-absorbing substances into the medium . The permeability barrier to Ca2+ was also damaged, and there was a release of radioactivity from bacteria labelled with 45Ca2+ . These events occurred at the optimum growth temperature, and took place at increased rates at higher temperatures . Damage was prevented to about the same extent by 0.1 mM-CaC12, BaC12 or MgC12 and by 10 mM-NaC1, KC1 or LiC1 . Ion competition experiments showed that Ca2+ was preferentially taken up and retained in comparison with Ba2+, Mg2+ and Na+, in that order . Isolated envelopes gave similar results . The dry weight of envelopes was reduced by 35% when they were suspended in water at 40 degrees C . It is clear that the function of certain envelope components in Achromobacter is highly dependent on divalent cations; and that both the integrity of the permeability barrier and the stability of the envelope are affected at low ion concentrations.

J Clin Pathol, 1976 Jun, 29(6), 537 - 42
Isolation of oxidase-positive Gram-negative cocci not belonging to the genus Neisseria from the urogenital tract; Platt DJ et al.; In a 12-month period, oxidase-positive, Gram-negative cocci showing similar characteristics in biochemical tests have been isolated from the urogenital tract of 39 male and female patients . Although these organisms superficially resemble Neisseria gonorrhoeae, biochemical characterization and the results of DNA base composition analysis indicate that they do not belong to the genus Neisseria . The relationship of these organisms to the genera Neisseria, Achromobacter, and Pseudomonas is discussed.

Br Med J, 1976 May 29, 1(6021), 1317 - 9
Bacterial glutaminase in treatment of acute leukaemia; Spiers AS et al.; A glutaminase-asparaginase enzyme from Achromobacter sp has antitumour activity in vitro and in animals . Glutaminase was administered in doses of 3500-20 000 IU/m2 body surface area/day to six patients with acute lymphoblastic leukaemia (ALL) and three patients with acute myeloid leukaemia (AML) . The enzyme had a blood half life of 80 minutes but depletion of blood glutamine persisted for 12 hours after single doses . Seven patients, including four (two with AML and two with ALL) resistant to asparaginase, received repeated doses of glutaminase . Antileukaemic effects were observed in all seven; one elderly patient developed metabolic acidosis . Study of this new antileukaemic agent in patients with acute leukaemia at an earlier stage of their disease is now justified.

Mikrobiologiia, 1976 May-Jun, 45, 475 - 80
{Biosynthesis of chitinase by Achromobacter liquefaciens}; Chigaleichik AG et al.; Bacterial cultures under study synthesize exocellular chitinase on a medium containing chitin or demineralized crab shells as a source of carbon and nitrogen . Conditions for biosynthesis of chitinase by the cells of Achromobacter liquefaciens 301a were investigated under periodic and continuous conditions of cultivation . The preparation of chitinase isolated from the cultural broth of A . liquefaciens 301a hydrolysed colloid and native chitin at the optimum pH 6.5 and temperature 40degreesC . The terminal products of the reaction are chitobiose and N-acetylglucosamine.

Biochim Biophys Acta, 1976 Mar 11, 429(1), 239 - 51
Chemical characterization and study of the autodigestion of pure collagenase from Achromobacter iophagus; Keil-Dlouha V; Only one collagenase (EC 3.4.24.3) is produced by the non-pathogenic Achromobacter iophagus strain . The chromatography of the crude enzyme on DE-32 cellulose followed by gel filtration on Sephadex G-100 in the presence of 1 M sodium chloride led to the isolation of a homogeneous enzyme . Its specific activity (1.642 mukat/mg) represents the highest value ever obtained for a bacterial collagenase . The amino acid composition of A . iophagus collagenase differs from that of Clostridium histolyticum mainly in the sulfur-containing amino acids . 1 mol of zinc was found for 1 mol of enzyme of molecular weight 104 000 . The autodegradation of the A . iophagus collagenase results in the formation of at least three active fractions which can be separated by preparative polyacrylamide gel electrophoresis as well as rechromatography on DE-32 cellulose . They are active towards the synthetic substrate as well as towards the native collagen . The results of ORD have shown that the digestion of the last one occurs in the helical parts of the substrate.

Ann Ist Super Sanita, 1976, 12(2-3), 93 - 112
Microbiological characteristics of natural mineral water; Schmidt-Lorenz W; Natural, non-carbonated mineral water is, like every other natural water from a spring, never sterile . However, the microbial level is always very low . But after its bottling, the level rises rapidly and numbers of more than 10,000 to 100,000/ml can be reached . In principle 2 groups of bacteria of very different origin and properties can be found in the microbial flora of the bottled, non-carbonated mineral water . Allochthonous bacteria will get into the water by contamination from the containers, closures, air or the bottling machines . They are mostly transitory as they cannot grow in a substrate with an extremely low nutritive level and die off more or less rapidly . From the hygienic point of view the permanently contaminating flora with Pseudomonas aeruginosa as main representative is more serious . These special gram-negative bacteria are oligocarbotolerant and can therefore multiply in the mineral water of extremely low nutrient level after a certain adaptation . Their effective bacteriological control is possible by colony counting with incubation at +37 degrees C but only just after bottling . The autochthonous microbial flora consists of psychrotrophic and of distinctly oligocarbophilic, mainly gram-negative bacteria such as Achromobacter, Flavobacteria, Pseudomonas as well as gram-positive Arthrobacter-species . According to indirect experiences, this autochthonous microbial flora must be growing in the open system of the underground source and renew itself constantly . The bottling of the natural spring water implies a drastic environmental change from this open system into a closed one . Then the bacteria start multiplying more or less rapidly like in a batch culture . Main reason for this is the extension of the inners surface of the system . The multiplication of bacteria after bottling of a mineral water of extremely low nutrient level therefore is an entirely normal biological process . For this reason, limits of the aerobic colony count at +20 degrees C incubation for natural mineral water seem not to be justified.

Enzyme, 1976, 21(2), 127 - 36
Determination of D-3-hydroxybutyrate dehydrogenase in mouse pancreatic islets with a photokinetic technique using bacterial luciferase; Berne C; A sensitive assay for d-3hydroxybutrate dehydrogenase (EC 1.1.1.30) was developed for use with the minute amounts of material obtained from islets of Langerhans microdissected from freeze-dried pancreatic sections . NADH formed in the enzyme reaction was determined by photokinetic analysis of the luminescence obtained with bacterial luciferase from Achromobacter fishcherii . In this way, accurate determination was obtained with less than 0.1 mug dry weight of islet material . In obese hyperglycemic mice, the islet enzyme had an activity of 4.7 mumoles/min and g dry weight . Optimal enzyme activity was found at pH 8 for the islet enzyme . The enzyme activity was similar in pancreatic islets and acini, while considerably higher activity was found in cardiac muscle, liver and renal cortex . Normal mouse islets showed about equal enzyme activity as the islets from obese hyperglycemic mice.

Eur J Biochem, 1975 Dec 1, 60(1), 267 - 9
Decreased dissociation of the 3-methylcrotonyl-CoA carboxylase complex from Achromobacter in the presence of 3-methylcrotonyl-CoA . A possible regulatory mechanism for the intracellular degradation of the enzyme; Schiele U et al.; By inactivation of different concentrations of 3-methylcrotonyl-CoA carboxylase from Achromobacter IVS with a fixed concentration of iodoacetamide, it was demonstrated that the degree of dissociation of the complex is considerably lower in the presence of 3-methylcrotonyl-CoA . ATP did not produce this effect . This property could serve to regulate the intracellular degradation of the enzyme, if the dissociated subunits were attacked preferentially.

Eur J Biochem, 1975 Dec 1, 60(1), 259 - 66
Investigations of the structure of 3-methylcrotonyl-CoA carboxylase from Achromobacter; Schiele U et al.; It was shown by gel electrophoresis in sodium dodecylsulphate solution that 3-methylcrotonyl-CoA carboxylase from Achromobacter IVS is composed of two different subunits with molecular weights of about 78000 and 96000, respectively . The biotin is bound to the heavier subunit . It was previously found that 3-methylcrotonyl-CoA carboxylase contains four biotin molecules per complex . A complex composed of four of each subunit would thus have a molecular weight of about 700000 . This is compatible with the molecular weight of 760000 determined earlier by analytical ultracentrifugation . Both subunits were isolated preparatively . As the subunits, unlike the complex, are very sensitive to oxygen, special precautions had to be taken during isolation . The biotin-containing subunit was isolated by chromatography on DEAE-cellulose in 5 M urea . It no longer catalyzed the overall reaction, yet could still carboxylate free biotin . The biotin-free subunit was separated after dissociation of the enzyme by three-days' dialysis at pH 9.8 under nitrogen . On chromatography over a Sepharose-bound avidin column, the biotin-subunit was fixed and the biotin-free subunit was eluted unretarded . The latter subunit showed no enzymic activity . After the addition of the biotin-containing subunit, overall activity was regenerated . The speed of reassociation is very much enhanced by 3-methylcrotonyl-CoA . It was shown by reassociation experiments under different conditions that probably an initial complex, AxBy is formed, possessing a binding site for 3-methylcrotonyl-CoA . Upon the binding of this substrate the conformation may be changed to a form favourable for reconstitution . Finally, the structures of biotin enzymes from different sources are compared . In the course of evolution there is a tendency toward integration of the different constituent proteins into only one polypeptide chain.

Appl Microbiol, 1975 Aug, 30(2), 242 - 50
Characterization of radiation-resistant vegetative bacteria in beef; Welch AB et al.; Ground beef contains numerous microorganisms of various types . The commonly recognized bacteria are associated with current problems of spoilage . Irradiation, however, contributes a new factor through selective destruction of the microflora . The residual microorganisms surviving a nonsterilizing dose are predominantly gram-negative coccobacilli . Various classifications have been given, e.g., Moraxella, Acinetobacter, Achromobacter, etc . For a more detailed study of these radiation-resistant bacteria occurring in ground beef, an enrichment procedure was used for isolation . By means of morphological and biochemical tests, most of the isolates were found to be Moraxella, based on current classifications . The range of growth temperatures was from 2 to 50 C . These bacteria were relatively heat sensitive, e.g., D10 of 5.4 min at 70 C or less . The radiation resistance ranged from D10 values of 273 to 2,039 krad . Thus, some were more resistant than any presently recognized spores . A reference culture of Moraxella osloensis was irradiated under conditions comparable to the enrichment procedure used with the ground beef . The only apparent changes were in morphology and penicillin sensitivity . However, after a few subcultures these bacteria reverted to the characteristics of the parent strain . Thus, it is apparent that these isolates are a part of the normal flora of ground beef and not aberrant forms arising from the irradiation procedure . The significance, if any, of these bacteria is not presently recognized.

J Biochem (Tokyo), 1975 Aug, 78(2), 355 - 61
Achromobacter cycloclastes nitrite reductase . The function of copper, amino acid composition, and ESR spectra; Iwasaki H et al.; 1 . Dialysis against cyanide at pH 7 of Achromobacter cycloclastes nitrite reductase {EC 1.7.99.3} of a dissimilatory type led to the removal of about 50% of the copper from the enzyme molecule, with a concomitant decrease of the enzymatic activities . It was inferred that enzyme-bound copper atoms play an essential role in the catalytic activities of the enzyme . 2 . The amino acid composition of the enzyme was determined after acid hydrolysis . 3 . ESR spectra of the frozen solution and lyophilized powder of the nitrite reductase predominantly showed the presence of two kinds of copper: Type 1 Cu2+, which had narrow and sharp hyperfine splitting, and Type 2 Cu2+, which had broader hyperfine splitting . The bond between the oxidized enzyme and nitrite seems to be ionic.

Eur J Biochem, 1975 Jun 16, 55(1), 263 - 9
Crystalline L-histidine ammonia-lyase of Achromobacter liquidum . Crystallization and enzymic properties; Shibatani T et al.; Crystalline L-histidine ammonia-lyase of Achromobacter liquidum was prepared with a 24% recovery of the activity . The specific activity of the pure enzyme (63 mumol of urocanic acid min-1 mg-1) is similar to those so far reported for the enzyme from other sources . The purified enzyme appeared to be homogeneous by analytical disc electrophoresis and isoelectric focusing (pI = 4.95) . The molecular weight determined by Sephadex G-200 gel filtration is 200000 . The optimum pH is 8.2, and the optimum temperature is 50 degrees C . The enzyme showed strict specificity to L-histidine (Km = 3.6 mM) . Several histidine derivatives are not susceptible to the enzyme but do inhibit the enzyme activity competitively; the most effective inhibitors are L-histidine methyl ester (Ki = 3.66 mM) and beta-imidazole lactic acid (Ki = 3.84 mM) . L-Histidine hydrazide (Ki = 36 mM) and imidazole (Ki = 6 mM) noncompetitively inhibited the enzyme EDTA markedly inhibited enzyme activity and this inhibition were reversed by divalent metal ions such as Mn2+, Co2+ Zn2+, Ni2+, Mg2+, and Ca2+ . These results suggest that the presence of divalent metal ions is necessary for the catalytic activity of histidine ammonia-lyase . Sodium borohydride and hydrogen peroxide inhibited the enzyme activity.

J Gen Microbiol, 1975 May, 88(1), 86 - 92
Unstable generalized transduction in Achromobacter; Woods DR et al.; Six auxotrophic markers of a halotolerant collagenolytic strain of Achromobacter were transduced by four alpha hages . Abortive transduction was also demonstrated . The generalized transduction system is unusual as the transductants were unstable, characteristic of transduction by lysogeny . The Achromobacter strain is a cryptic lysogen for alpha and purified transductants were either sensitive or resistant to alpha . Purified clones from four resistant transductants and one sensitive transductant liberated phage spontaneously . The host ranges of these spontaneous phage differed from that of the alpha phage used for the transduction experiment . Some initially resistant transductants became simi-sensitive to alpha (efficiency to plating) e.o.p . (10minus-1 to 10minus-2) after repeated cloning.

Z Naturforsch {C}, 1975 May-Jun, 30(3), 425 - 6
Over-production of porphyrins and heme in heterotrophic bacteria; Philipp-Dormston WK et al.; In Escherichia coli, Pseudomonas aeruginosa, and Achromobacter metalcaligenes gamma-aminolevulinic acid synthase and gamma-aminolevulinic acid dehydratase appear to be the rate limiting enzymes of porphyrin and heme biosynthesis . Bypassing of these enzymes by addition of gamma-aminolevulinic acid or porphobilinogen results in overproduction of tetrapyrroles.

Ann Microbiol (Paris), 1975 Apr, 126(3), 367 - 80
{On the taxonomy and physiology of bacteria utilizing hydrocarbons in the sea (author's transl)}; Le Petit J et al.; Using mineral media with gas oil as sole carbon source, 191 bacterial strains were isolated from the costal area of Marseille . These strains were attributed to Achromobacter, Alcaligenes, Pseudomonas, Acinetobacter and Arthrobacter genera . Amongst isolated strains there was a predominance of the Alcaligenes-Achromobacter group over others genera . Growth and respiratory activity of 5 strains were studied on hexadecane and acetate . Respiratory activity on hexadecane of 32 strains cultured on acetate has been measured . These strains could be placed in two groups . The group 1 shows an immediate respiratory activity which is not abolished by chloramphenicol . The group 2 presents either an immediate or a delayed respiratory activity which is always abolished by chloramphenicol . Maintenance or suppression of respiration by chloramphenicol is a character which is homogenously distributed amongst the species.

Pathol Biol (Paris), 1975 Apr, 23(4), 277 - 82
{A hospital epidemic due to Achromobacter calcoaceticus}; Lecocq E et al.; 35 patients were secondarily infected in our hospital with a strain of A . calcoaceticus resistant to the usual antibiotics and sulfonamides, but sensitive to colimycin . The epidemic lasted 118 days and is not yet finished . Each of the infected patients had a severe surgical or medical illness, underwent operations, trachetomy, etc . and was treated with antibiotics . A . calcoaceticus persisted alone or was associated with other bacteria from 1 to 46 days, in specimens (sputum, etc . or in blood) sometimes until death . It is not pathogenic for rabbits and mice; its pathogenicity in man is discussed in the text . The epidemic strain was not harboured by 15 doctors, students or nurses, nor present on 147 objects in the vicinity of the patients, but was found in a bottle of aqueous 1% eosin solution in the room of an infected child . Experiments show that A . calcoaceticus is not killed by a 1% eosin solution, and freely multiplies in broth containing 0,5% eosin . It is easy to identify the first case and chronology of the epidemic, but it is less easy to identify each time the means of propagation . Usually, infected patients seem to be the source of further infection . Does eosin paly more than an occasional role? Were the germs transported by dust? This not very disturbing epidemic suggests that less pathogenic germs may still cause unsuspected hospital epidemics.

Can J Ophthalmol, 1975 Apr, 10(2), 290 - 2
Clostridium welchii corneal ulcer--a case report; Majekodunmi S et al.; A case of corneal ulcer in a 10-year-old Nigerian boy in which clostridium welchii and achromobacter were isolated is reported . The absence of trauma and rarity of only corneal involvement in a clostridial infection is emphasized . The bacteriology of the clostridial infection is emphasized . The bacteriology of the clostridial ocular infections is reviewed, the media necessary for isolation of the organism and the symbiotic relationship with other bacteria are discussed . The clinical course of the disease with uneventful recovery of the patient is emphasized.

Biochim Biophys Acta, 1975 Mar 28, 384(1), 228 - 34
Collagenase production by Achromobacter iophagus; Welton RL et al.; Achromobacter iophagus produced collagenase (EC 3.4.24.3) when cultured aerobically in buffer containing 5% peptone . The bacterium is non-pathogenic and tests on rabbits indicated that the culture medium was atoxic . The collagenase, which hydrolyzed insoluble and soluble native collagen, was purified by (NH4)2 SO4 precipitation, starch block electrophoresis, and gel filtration . It was shown to be serologically distinct from Clostridium histolyticum collagenase and to have molecular weight and sedimentation coefficient values of approx . 112 000 and 5.3 S, respectively.

Bull Environ Contam Toxicol, 1975 Feb, 13(2), 249 - 55
Bacterial degradation of polychlorinated biphenyls II . Rate studies; Wong PT et al.; Polychlorinated biphenyls (Aroclor 1221, 1242 and 1254) at concentrations up to 0.1% in glucose did not inhibit the growth of lake water bacteria . The bacteria used Aroclor 1221 and 1242 but not 1254 as sole carbon and energy sources for growth . Less than 1% of lake water bacteria, however, possess this ability . Seven bacterial isolates from Aroclor agar plates were identified; five belonged to Achromobacter sp . and two were Pseudomonas sp . The metabolic breaksown of Aroclor 1221 was followed . THE MIXTURE WAS COMPLETELY DEGRADED INTO SEVERAL LOW MOLECULAR WEIGHT COMPOUNDS AFTER ONE MONTH INCUBATION . Unchlorinated biphenyl was degraded at a faster rate than 2-chlorobiphenyl and 4-chlorobiphenyl isomers.

Mikrobiologiia, 1975 Jan-Feb, 44(1), 76 - 80
{Decomposition of cholesterol by Achromobacter candicans}; Imsenecki AA et al.; The decomposition of cholesterol by the cell suspensions of Achromobacter candicans 42 and by the cell-free extracts of this bacterium was studied . The decomposition of cholesterol in the presence of the wet biomass during four hours of the incubation was 72.5 percent, and with the cells that were preliminarily lyophilized or treated by acetone 49.8 and 23.0 percent, respectively . The activity decreased from 70.0 to 50.0 percent if the biomass was kept at minus 3 degrees C during 70 days . The decomposition of cholesterol by the cell-free preparations (supernatant and protein after precipitation with ammonium sulphate to 0.7 saturation) constituted 40.0 percent of the cholesterol contained in a sample . One of the intermediate products of cholesterol decomposition was delta4-cholesten-3-one.

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