Biotechnol Adv, 1993, 11(3), 525 - 37 Development of high penicillin producing strains for solid state fermentation; Barrios-Gonzalez J et al.; Penicillin production with an industrial strain and 4 strains of P . chrysogenum, in solid state fermentation (SSF) and liquid submerged fermentation (LSF), was determined . Their ability to produce the antibiotic in SSF in relation to their capacity to do so in LSF was evaluated . this was done by calculating the ratio PS/PL (production in SSF/production in LSF), which was called relative production . Clones were isolated from each strain and evaluated in a similar way . The strains presented different relative productions (from 1.4 to 2.5) . Within the clones, a much wider range of relative productions was observed (0.6 to 16.7) . On the other hand, the highest-producing strains in LSF were also the highest producers in SSF . This indicates that the production potential of a strain is an important factor in its production level in SSF . Moreover, the highest penicillin producing ciones (9,500 to 10,500 microg of penicillin/g were generated from high-yielding strains (P2 and ASP-78) . However, the higher-producing strains (in LSF) showed lower relative performance, suggesting that higher producing strains tend to express less efficiently their potential in SSF . In this study, several overproducing clones, particularly suited for SSF, were obtained by the procedures followed . Production increases of 500 to 600 %, in this culture system, were achieved. Biotechnol Adv, 1993, 11(3), 519 - 23 Caffeine reduction in coffee pulp through silage; Porres C et al.; Silage tests to study reductions of antiphysiological compounds (caffeine and polyphenols) of fresh coffee pulp during the anaerobic fermentation were done . A concrete silo divided in compartments, with a total capacity of 9 tons of fresh material was utilized . The silage periods ranged between 99-224 days and the following materials were ensiled: 1) coffee pulp, 2) coffee pulp with sugar cane molasses, 3) coffee pulp with a mixture of molasses and ammonia and 4) screw pressed coffee pulp with molasses . Reductions in caffeine, total polyphenols and condensed polyphenols ranged between 13-63%, 28-70% and 51-81% respectively.It was concluded that in the case of coffee pulp, silage presents and ideal method to preserve the material and partially reduce the contents of antiphysiological compounds. Biotechnol Adv, 1993, 11(3), 509 - 17 Bioreduction of prochiral ketones with yeast cells cultivated in a vibrating air-solid fluidized bed fermentor; Matsuno R et al.; A brief review of fluidized bed fermentors and of bioreduction of prochiral ketones by yeast cells is presented . Cultivation of yeast cells, Saccharomyces cerevisiae HUT 7099, in a vibrating fluidized bed and the bioreduction of ethyl acetoacetate by the cells are described . The cultivation of the cells in the fermentor was successfully performed at relatively low moisture content, about 40 % on wet basis . The cell size decreased and the shape changed from ellipsoid to spherical after the logarithmic growth phase . The biocatalytic performance of yeast cells cultivated in submerged, static solid, and fluidized bed cultures was compared . The cells cultivated in static solid culture exhibited the highest activity . Possible accumulation of energy sources by the cells was suggested as the explanation for better performance. Biotechnol Adv, 1993, 11(3), 495 - 505 Protein enrichment of sweet potato residue with co-culture of amylolytic fungi by solid-state fermentation; Yang SS; Protein enrichment of sweet potato residue with amylolytic moulds by solid-state fermentation was higher than that obtained with amylolytic yeasts . The optimum initial moisture content for protein enrichment was 66% to 75% . Incrementally added nitrogen sources to the culture at zero time and at 24 h considerably improved the final protein content . During the cultivation, the moisture, ash and ATP contents increased, while the pH value decreased . A 1:1 co-culture of amylolytic mycelial fungi yielded a product with 32.4% crude protein after 4 days incubation at 30 degrees C. Biotechnol Adv, 1993, 11(3), 469 - 79 Fermentation of cellulosic materials to mycoprotein foods; Moo-Young M et al.; A new bioprocess is described in which a cellulolytic, food-grade fungus Neurospora sitophila converts cellulosic materials to protein-rich products for food and fodder . The optimal conditions for the conversion are identified: 35-37 degrees C temperature, pH 5.5, 2.35 ms(-1) agitator tip speed . Scale-up of the production process to 1,300 L is reported . The mycoprotein production data on several types of cellulosic materials (sugarcane bagasse, corn stover, wood cellulose) are presented . The performance of N . sitophila is found to compare favourably with that of Chaetomium cellulolyticum, another cellulolytic organism previously reported on by us. Biotechnol Adv, 1993, 11(3), 441 - 65 Properties of thermostable hemicellulolytic enzymes from Thermomonospora strain 29 grown in solid state fermentation on coffee processing solid waste; Srivastava KC; During decaffeination of Coffee Processing Plant Solid Wastes (CPSW) by actinomycetes, Thermomonospora, Strain 29 exhibited high titers of cellulase and xylanase . This organism, originally isolated on soybean seed coat was grown in solid state fermentation on CPSW supplemented with mineral salts . Enzymes recovered were arabinosidase, xylanase, and beta-D-xylosidase . Higher activity of the former two enzymes was in the extracellular broth, whereas the beta-D-xylosidase activity was highest in the cell fraction . The enzymes were characterized after precipitation with (NH(4))(2)SO(4), dialysis, and gel filtration . Production of all three enzymes was inhibited by monomeric sugars and sugar alcohols but not by arabinoxylan, xylans, or xylan containing water insoluble carbohydrates . The optimum pH for the activity was 6.5, 7.0, and 7.5 for beta-xylosidase, xylanase and arabinosidase (alpha-L-arabinofuranosidase, alpha-arabinosidase, alpha-L-arabinosidase) respectively . These enzymes were stable in the pH range of 6.5 to 8.0 . All three enzymes were thermostable up to 80 degrees C . At 55 degrees C, arabinosidase had the longest half life of 120 h . However, at 40 degrees C, xylanase had the longest half life (504 h) . At either temperature, beta-D-xylosidase had the shortest half life . The molecular weights (kDa), and Kms (mM) were estimated to be 95, 0.27; 45, 12.4; and 106, 0.67 for arbinosidase, xylanase, and beta-xylosidase respectively . Step wise addition of the three enzymes showed higher saccharification of lignocellulosics. Biotechnol Adv, 1993, 11(3), 429 - 40 New approach for selecting pectinase producing mutants of Aspergillus niger well adapted to solid state fermentation; Antier P et al.; The aim of this paper is to review and study a new approach for improving strains of Aspergillus niger specially adapted to produce pectinases by Solid State Fermentation (SSF) with materials having low levels of water activity (a(w)), i.e., coffee pulp . Special emphasis is placed on the use of two antimetabolic compounds: 2-deoxy-glucose (DG) and 2,4-dinitro-phenol (DNP) combined with a water depressant (ethylene glycol = EG) in order to put strong selection pressures on UV treated spores from parental strain C28B25 isolated from a coffee plantation . Such a strain was found to be DG sensitive . Results suggested the existence of a reciprocal relation between adaptation of isolated strains to SSF or to Submerged Fermentation (SmF) systems . Preliminary physiological analysis of isolated strains showed that at least some few initially DG resistant mutants could revert to DG sensitive phenotype but conserving increased pectinase production . Also it was found that phenotype for DNP resistance could be associated to changes of DG resistance . Finally, it was found that low levels of a(w) produced by adding 15% EG to agar plates, were a significant selection factor for strains well adapted to SSF system. Biotechnol Adv, 1993, 11(3), 417 - 27 Production of novel oligosaccharide oxidase by wheat bran solid-state fermentation; Lin SF et al.; A search for oxidases that catalyze the oxidation of oligosaccharides has resulted in the isolation of several soil-derived fungus strains which produced novel oligosaccharide oxidases with different substrate specificity on wheat bran solid culture . One of these oxidases produced by Acremonium strictum T1 strain has been characterized . This enzyme showed high reactivity toward maltose, lactose, cellobiose and maltooligosaccharides composed of up to seven glucose units, and was named as glucooligosaccharide oxidase based on its substrate specificity . Strain T1 was subjected to a strain improvement program, and an enzyme hyper-producing mutant strain T1-38 was selected . This mutant strain produced glucooligosaccharide oxidase 75 times higher than the wild type strain T1 . When cultivated in a solid medium comprised of 1 part of wheat bran and 1 part of water (w/w), enzyme activity reached a maximum level of 6 units per g of culture medium after 4 days cultivation . Characteristics of the enzyme including the substrate specificity were compared with two other novel oligosaccharide oxidases isolated in this laboratory . Batch type conversion of lactose to lactobionic acid using crude enzyme was also discussed. Biotechnol Adv, 1993, 11(3), 409 - 16 Effect of culture conditions on lipolytic enzyme production by Penicillium candidum in a solid state fermentation; Ortiz-Vazquez E et al.; Lipolytic enzymes were produced using wheat bran as substrate in a solid state fermentation with Penicillium candidum . The best production of lipolytic activity occurred at 29 degrees C . One hundred micromoles of free butyric acid (FBA) was released from tributyrin by 1 mL of cell free supernatant in the absence of control of environmental relative humidity . When a closed chamber saturated with water vapour was used the lipolytic activity increased to 320 micromoles of free butyric acid . The best initial reaction pH was 7.0 . The highest activity, 480 micromoles of FBA, was obtained at a moisture content of 67.5 % of saturation. Biotechnol Adv, 1993, 11(3), 399 - 407 Downstream processing of microbial rennet from solid state fermented moldy bran; Thakur MS et al.; In recent years due to acute shortage of calf-rennet, microbial rennets seem to be an effective alternative and are commercially produced . Mucor miehei was cultivated under the solid state fermentation conditions, and the moldy bran was extracted using a semicontinuous multiple contact forced percolation method . The treated extract was then filtered through 5% R16 clay which enabled easy and efficient removal of impurities such as lipase and protease without involving costly chemical treatments . The ethyl alcohol precipitated enzyme was dried and made into powder form having activity of 1.5 x 10(5)Soxhlet units/gm. Biotechnol Adv, 1993, 11(3), 387 - 97 Solid state fermentation: acid protease production in controlled CO2 and O2 environments; Villegas E et al.; The effect of the partial pressure of O(2) and CO(2) on the acid protease production in solid state fermentation by Aspergillus niger on wheat bran was studied . A fermentation system was used, which allowed on-line reactor measurements and continuous data acquisition of pH, temperature, gas flow, pressure drop and CO(2) production . Six paired combinations of CO(2) and O(2) concentrations were studied . The results showed a direct relationship between pressure drop, production of CO(2) and temperature increase . The pH evolution patterns were similar in all cases but different if the measurements were made on-line or on a liquid homogenate of the fermented substrate . Acid protease production was increased when the gas had 4% CO(2), (vol/vol), and it reached its highest level, a 43% increase over air, with a mixture of 4% CO(2) and 21% O(2) . The protease production was strongly related to the mold metabolic activity as represented by the total CO(2) evolved. Biotechnol Adv, 1983, 1(1), 47 - 58 Fuels and industrial chemicals; Linko P; The favorite subject of recent literature on biotechnical processes has been ethanol fermentation . This review covers a number of new technics developed, including immobilized biocatalyst technology and bacterial fermentations . Reference is also made to recent work on whey, starch, inulin, and cellulosic materials as substrates for ethanol production . Renewed interest in acetonebutanol fermentation for solvent and liquid fuel production has also been clearly evident during the last two years . Biotechnical production of organic acids has been considered as an alternative route to chemical feedstocks . New developments in amino acid, methane, hydrogen, and hydrocarbon production, and on hydrocarbon oxidation are also briefly covered. Biotechnol Adv, 1985, 3(2), 209 - 18 Biosensors: recent trends; Graham A et al.; One of the major bottlenecks in automation and process control of industrial bioprocesses is the lack of suitable sensing devices to accurately measure the concentrations of biomolecules . The measurement of ions (e.g., H(+), NH(4)(+)) and gases (e.g., O(2), CO(2), NH(3)) using standard ion-selective and gas sensing electrodes respectively, is well established . Chemical analysis of biomolecules off-line is generally unreliable, labour intensive and may lead to contamination of the biological systems . Problems of maintaining sterile conditions are especially important when dealing with slow growing mammalian or plant cells in culture . Active research in the development of biosensors for monitoring fermentation processes, food production and pollution control, and for medical and veterinary applications is currently underway . This paper reviews recent approaches toward the development of biosensors which involve a biochemical interaction to measure the concentrations of biomolecules, primarily for the on-line monitoring and control of fermentation processes. Biotechnol Adv, 1984, 2(1), 21 - 33 Effective diffusivity of oxygen in microbial pellets; Ho CS et al.; In a typical submerged aerobic fermentation with microbial pellets, the effective diffusivity of oxygen in the pellets is probably the most important, yet most difficult transport property to characterize experimentally . Its values directly indicate the efficiency or deficiency of oxygen to individual cells, and thus the biological activity of the microorganisms . In the past, it was not possible to assess reliably the effective diffusivity of oxygen in pellets due to several reasons . Firstly, most oxygen electrodes available were coarse, and hence not suitable for in situ measurements . Secondly, there was a lack of methods rigorous enough to characterize the structure of the microbial pellets . A state-of-the-art review of the literature relating to the feature subject is presented . Emphasis is laid upon development and evolution of the means for quantitative characterization of the effective diffusivity of oxygen in microbial pellets. Biotechnol Adv, 1992, 10(4), 577 - 95 Biotransformations of rifamycins: process possibilities; Banerjee UC et al.; Rifampicin, an important antibiotic, is manufactured by chemical conversion of rifamycin S which is obtained by the chemical modification of rifamycin B . Rifamycin B is a product of Nocardia mediterranei fermentations . The chemical conversion of rifamycin B to rifamycin S has many disadvantages: Strong acidic conditions are required, heavy foam formation accompanies transformation and the yields are low . This review highlights the developments in alternative, biochemical transformations using enzymes and cells; the main focus is on transformations carried out by rifamycin oxidase. Biotechnol Adv, 1990, 8(4), 685 - 97 Regulation of D-xylose utilization by hexoses in pentose-fermenting yeasts; Webb SR et al.; The aldopentose D-xylose is one of the most abundant sugars in plant biomass and its efficient microbial utilization is of fundamental importance in the overall bioconversion of lignocellulosic materials into liquid fuels and chemicals . The discovery of pentose-fermenting yeasts in the early 1980's led to world wide interest because of the perceived potential for improved D-xylose fermentation to enhance the prospect of biomass conversions . However, the utilization of D-xylose by pentose-fermenting yeasts can be adversely affected by the hexoses, mainly D-glucose and D-mannose, which are usually present in high amounts in lignocellulosic hydrolysates . Research in the past several years has uncovered some of the regulatory effects of D-glucose on D-xylose utilization . However, much remains unknown about the mechanisms responsible for these effects . This review summarizes the current state of knowledge on the induction, repression and inactivation of D-xylose utilization in pentose-fermenting yeasts. Biotechnol Adv, 1989, 7(4), 467 - 88 Downstream processing of proteins: recent advances; Ogez JR et al.; This review on the downstream processing of proteins describes innovations that have occurred in the field since 1983 . Several areas have seen particularly high levels of achievement, and are accorded expanded coverage relative to our previous review {1} . As an example, the increasing integration of downstream operations with upstream technologies, such as molecular biology and fermentation, has led to the development of some very powerful processes . The degree to which organizations understand that there needs to be one unified process, rather than the independent steps of cloning, fermentation and recovery, seems directly related to the ultimate speed and success of the development effort.In 1983 one of the most active development areas was chromatography, especially affinity chromatography . This is still true today, and this topic has been expanded to include biospecific adsorptions that would not traditionally be classified as chromatography . With more proteins being developed for human administration, there has been an increased emphasis on all aspects of process hygiene . In addition, there has been much discussion about the impact of regulatory demands on the design and development of the manufacturing processes . Therefore, a section has been added which covers several of the regulatory issues that have been raised for products of the new biotechnology . Finally, as some of the early process development achievements are now beginning to bear fruit in the form of patents, we have increased our citation of this area of the literature. Biotechnol Adv, 1986, 4(2), 279 - 88 Production of plant secondary metabolites without plants: a perspective; Ellis BE; Efforts to commercially exploit native plant secondary metabolite production patterns in cell culture systems have been largely thwarted by the repression of secondary metabolism under growth-oriented culture conditions . Where expression can be obtained by selection or elicitation, the difficulties of large scale cultivation/processing still make the cost effectiveness of cell culture systems dubious except where a very high value market niche can be identified.The long range prospect for efficiently utilizing the catalytic genius of higher plants resides in the transfer of appropriate genetic information to microbial systems for whom the fermentation expertise and industrial facilities already exist. Biotechnol Adv, 1986, 4(2), 219 - 43 Fermented foods, feeds, and beverages; Steinkraus KH; World population reached 5 billion people in 1986 and is expected to rise to 6 particularly severe in parts of Africa, particularly Ethiopia . Hunger and malnutrition accompany the poverty that is characteristic of the masses of the people in the developing world . Even in the United States, one of the most affluent countries, there has been a considerable increase in the number of homeless people including mothers and children living in the streets or in substandard housing . The indigenous fermented foods and beverages already consumed for centuries by hundreds of millions of the world's masses can be used in many cases to improve and extend the world's food and nutrition supply at a relatively low cost . Fermented foods, feeds and beverages are getting ever increasing attention particularly in the developing world and also in the developed world . This paper summarizes activities and advances related to fermented foods, feeds, and beverages over the past several years. Biotechnol Adv, 1989, 7(1), 1 - 14 Enhancement of oxygen transfer rates in fermentation using oxygen-vectors; Rols JL et al.; Oxygen transfer is one of the bottlenecks in conventional fermentation technology and it has so far been almost totally overlooked with regards to high cell densities and immobilized cells . This review presents some new concepts to improve oxygen supply in aerobic fermentations, especially the use of oxygen-vectors . The oxygen-vectors generally used are liquids which are insoluble in the fermentation media . Their utilization in an emulsified form can significantly increase the oxygen transfer coefficient between gas and aqueous phases . It seems that the vector acts as an active intermediate in the oxygen transport from gas bubbles to aqueous phase, but the mechanisms involved in this unconventional technique of aeration are not yet known. Biotechnol Adv, 1991, 9(4), 623 - 41 Regulation of ACV synthetase in penicillin- and cephalosporin-producing microorganisms; Zhang J et al.; ACV synthetase is the first enzyme in the biosynthetic pathway for all natural penicillins and cephalosporins . Its activity catalyzes the possible rate-limiting step and is subject to various regulatory controls . In both the fungus Cephalosporium acremonium and the actinomycete Streptomyces clavuligerus, formation of the enzyme is repressed by ammonium and phosphate ions, but not by easily-utilized carbon sources; it is induced by methionine in C . acremonium . The action of the crude enzyme is indirectly inhibited in vitro by sugars such as glucose and by the carbon source metabolite glyceraldehyde-3-phosphate (G3P) . Sugars are not inhibitory to the purified enzyme activity but G3P is inhibitory . The sugar inhibition is reversed by ATP and the G3P inhibition by L-cysteine (L-cys) . Addition of L-cys to fermentation media increases beta-lactam production by both microorganisms . Phosphate and ferrous ions inhibit enzyme activity . Dissolved oxygen levels do not affect enzyme formation . Regulation of ACVS formation most likely occurs at the transcriptional level. Biotechnol Adv, 1991, 9(4), 559 - 612 On the merits of viable-cell immobilisation; Dervakos GA et al.; Many advantages have been claimed over the years for the use of immobilised cells, both as enzyme systems and as whole viable cell systems for complete fermentation reactions . However, few of the claims have been fully substantiated, and may not even be entirely justified . Most research is involved with single applications and the best that can be hoped for is some evidence that immobilised cells in each of these individual cases display some advantage over the equivalent free cell system . The purpose of this review is to assess the merits of viable cell immobilisation in the light of published literature and to elucidate the underlying mechanisms . Particular attention is paid to the generally unanticipated, but widely observed enhanced stability of immobilised cell fermentation processes. Biotechnol Adv, 1983, 1(2), 247 - 61 Downstream processing of proteins; Becker T et al.; This review focuses on the recovery of proteins from fermented starting materials, covering developments that have appeared in the literature since 1979 . The major subjects discussed include cell disruption and extraction, solid/liquid separations, chromatography, separations in solution, and finishing operations . While advances have occurred in every area, the greatest diversity of innovation has taken place in the fields of chromatography and tangential-flow filtration. Biotechnol Adv, 1999 Apr, 17(1), 29 - 48 Control of fed-batch fermentations; Lee J et al.; Fed-batch fermentation is used to prevent or reduce substrate-associated growth inhibition by controlling nutrient supply . Here we review the advances in control of fed-batch fermentations . Simple exponential feeding and inferential methods are examined, as are newer methods based on fuzzy control and neural networks . Considerable interest has developed in these more advanced methods that hold promise for optimizing fed-batch techniques for complex fermentation systems. Biotechnol Adv, 1999 Nov, 17(6), 477 - 88 Applications of neural networks to recovery of biological products; Patnaik PR; Artificial neural networks (ANN) are being applied to recovery of products from fermentation broths . Recovery methods for which mathematical models are complex or non-existent are particularly suitable for control and analysis by ANNs . Use and potential of artificial neural networks for product recovery applications are reviewed. Biotechnol Adv, 2000 May, 18(3), 207 - 17 Production, purification, and characterization of the debittering enzyme naringinase; Puri M et al.; This review discusses the debittering enzyme naringinase and its essential role in the commercial processing of citrus fruit juice . Applications of this enzyme in other areas are identified . Characterization of the enzyme is detailed and its immobilized preparations are discussed . Production of microbial naringinase by fermentation is described. Biotechnol Adv, 2000 Aug, 18(5), 403 - 23 Alpha-L-arabinofuranosidases: biochemistry, molecular biology and application in biotechnology; Saha BC; Interest in the alpha-L-arabinofuranosidases has increased in recent years because of their application in the conversion of various hemicellulosic substrates to fermentable sugars for subsequent production of fuel alcohol . Xylanases, in conjunction with alpha-L-arabinofuranosidases and other accessory enzymes, act synergistically to degrade xylan to component sugars . The induction of alpha-L-arabinofuranosidase production, physico-chemical characteristics, substrate specificity, and molecular biology of the enzyme are described . The current state of research and development of the arabinofuranosidases and their role in biotechnology are presented. Biotechnol Adv, 2000 Oct, 18(6), 499 - 514 Small bugs, big business: the economic power of the microbe; Demain AL; The versatility of microbial biosynthesis is enormous . The most industrially important primary metabolites are the amino acids, nucleotides, vitamins, solvents, and organic acids . Millions of tons of amino acids are produced each year with a total multibillion dollar market . Many synthetic vitamin production processes are being replaced by microbial fermentations . In addition to the multiple reaction sequences of fermentations, microorganisms are extremely useful in carrying out biotransformation processes . These are becoming essential to the fine chemical industry in the production of single-isomer intermediates . Microbially produced secondary metabolites are extremely important to our health and nutrition . As a group, they have tremendous economic importance . The antibiotic market amounts to almost 30 billion dollars and includes about 160 antibiotics and derivatives such as the beta-lactam peptide antibiotics, the macrolide polyketide erythromycin, tetracyclines, aminoglycosides and others . Other important pharmaceutical products produced by microrganisms are hypocholesterolemic agents, enzyme inhibitors, immunosuppressants and antitumor compounds, some having markets of over 1 billion dollars per year . Agriculturally important secondary metabolites include coccidiostats, animal growth promotants, antihelmintics and biopesticides . The modern biotechnology industry has made a major impact in the business world, biopharmaceuticals (recombinant protein drugs, vaccines and monoclonal antibodies) having a market of 15 billion dollars . Recombinant DNA technology has also produced a revolution in agriculture and has markedly increased markets for microbial enzymes . Molecular manipulations have been added to mutational techniques as means of increasing titers and yields of microbial procresses and in discovery of new drugs . Today, microbiology is a major participant in global industry . The best is yet to come as microbes move into the environmental and energy sectors. Biotechnol Adv, 2000 Nov 1, 18(7), 581 - 99 Integrated processing of biotechnology products; Schugerl K; Integrated bioprocessing in which a potentially inhibitory product is continuously removed from the fermentation broth as it is produced, has important advantages in improving yield and conversion relative to conventional processes . This review discusses integrated processing for ethanol, butanol, organic acids, antibiotics, and other products . A variety of recovery operations can be used to isolate the product, as discussed . Use of some of the available options is compared. Biotechnol Adv, 2001 Jun, 19(3), 201 - 23 Glycerol production by microbial fermentation: a review; Wang ZX et al.; Microbial production of glycerol has been known for 150 years, and glycerol was produced commercially during World War I . Glycerol production by microbial synthesis subsequently declined since it was unable to compete with chemical synthesis from petrochemical feedstocks due to the low glycerol yields and the difficulty with extraction and purification of glycerol from broth . As the cost of propylene has increased and its availability has decreased especially in developing countries and as glycerol has become an attractive feedstock for production of various chemicals, glycerol production by fermentation has become more attractive as an alternative route . Substantial overproduction of glycerol by yeast from monosaccharides can be obtained by: (1) forming a complex between acetaldehyde and bisulfite ions thereby retarding ethanol production and restoring the redox balance through glycerol synthesis; (2) growing yeast cultures at pH values near 7 or above; or (3) using osmotolerant yeasts . In recent years, significant improvements have been made in the glycerol production using osmotolerant yeasts on a commercial scale in China . The most outstanding achievements include: (1) isolation of novel osmotolerant yeast strains producing up to 130 g/L glycerol with yields up to 63% and the productivities up to 32 g/(L day); (2) glycerol yields, productivities and concentrations in broth up to 58%, 30 g/(L day) and 110-120 g/L, respectively, in an optimized aerobic fermentation process have been attained on a commercial scale; and (3) a carrier distillation technique with a glycerol distillation efficiency greater than 90% has been developed . As glycerol metabolism has become better understood in yeasts, opportunities will arise to construct novel glycerol overproducing microorganisms by metabolic engineering. Biotechnol Adv, 1995, 13(2), 209 - 34 Fungal production of citric acid; Grewal HS et al.; Citric acid is the principal organic acid found in citrus fruits . To meet increasing demands it is produced from carbohydrate feedstock by fermentation with the fungus Aspergillus niger and the yeasts of Candida spp . Effect of various fermentation conditions and the biochemistry of citric acid formation by A . niger have been discussed . Commercially citric acid is produced by surface, submerged and solid state fermentation techniques . Recovery of pure acid from fermentation broth is done primarily by precipitation with lime and also by solvent extraction. Biotechnol Adv, 1995, 13(2), 175 - 208 Liquid emulsion membranes: principles, problems and applications in fermentation processes; Patnaik PR; Liquid emulsion membranes (LEMs) have developed into a versatile technique for a variety of applications involving selective and controlled transport of biochemicals . Biological applications cover the controlled delivery of drugs from capsules, detoxification of the circulatory system, recovery of useful compounds from waste streams and selective separation of products from fermentation broths . This review traces the development of LEMs, discusses their key features, advantages and limitations, describes methods of modelling LEM systems and highlights some applications with industrial potential.Two kinds of LEM systems are considered . The first type are agitated emulsions, which are relatively easy to prepare and use but may be limited in their selectivity and long-term stability . Supported liquid membranes (SLMs) are a recent development; they use porous solid supports and have excellent stability and selectivity . Their chemical engineering aspects and applications in fermentation processes are considered. Biotechnol Adv, 1995, 13(3), 425 - 53 Plant cell and tissue culture: alternatives for metabolite production; DiCosmo F et al.; Plant cell culture systems represent a potential renewable source of valuable medicinals, flavours, essences and colourants that cannot be produced by microbial cells or chemical syntheses . However, only a few cultures produce these compounds in commercially useful amounts . The low productivities are associated with our poor understanding of the biochemistry of these systems . Recent advances in molecular biology, enzymology, physiology and fermentation technology of plant cell cultures suggest that these systems will become a viable source of important natural products . This review examines the sate of the art of production of medicinal plant secondary metabolites by plant cell cultures. Wei Sheng Yan Jiu, 2003 Jul, 32(4), 384 - 5 {Analysis of kojic acid in Aspergillus oryzae ferment by ion-pair reversed-phase high performance liquid chromatography}; Zhao S et al.; A new and an efficient method for analyzing kojic acid in aspergillus oryzae ferment by ion-pair reversed-phase high performance liquid chromatography was developed . The kojic acid was well separated on a C18 column using methanol-0.01 mol/L disodium hydrogen phosphate and 2 mmol tetrabutylammonium bromide (4:96) as the mobile phase, followed by an UV detection at 226 nm . The detection limit was 0.012 microgram/ml . The spiked recoveries in wort-yeast culture, wort-peptone culture and potato-yeast-sugar culture were 98.2%, 103.4% and 97.2%, respectively . The RSD values were 0.51%, 0.45% and 0.43% respectively. Arch Tierernahr, 2003 Aug, 57(4), 279 - 95 Effects of protozoa on methane production in rumen and hindgut of calves around time of weaning; Schonhusen U et al.; Effects of the presence or absence of ciliate protozoa on methanogenesis in the rumen and hindgut were investigated in young calves during a 7-week period . Ten Holstein calves, aged 7 days, were divided in two groups (n = 5) and fed an increasing amount of a commercial milk replacer and small amounts of a calves starter . One group was inoculated with ciliate fauna on two occasions, week 5 and 6, while the second remained ciliate-free . The absence of protozoa in the rumen decreased rumen empty weight (-23%, P < 0.01), and rumen pool size of N (-36%, P < 0.01) and crude fat (-37%, P < 0.05) . Rumen bacteria of non-faunated calves contained a higher proportion of total amino acid-N per 16 g N (+3%, P < 0.01) and D-alanine-N per 16 g N (+13%, P < 0.05) compared to faunated calves . Further results contain a reference for a higher bacterial mass in the ciliate-free rumen with an increased number of bacteria adherent to rumen mucosa . The CH4 production in the rumen increased exponentially with the increase in protozoa population size (R2 = 0.68) . In presence of 46 x 10(4) protozoa per ml rumen fluid, the in vitro CH4 production of rumen fluid per mol total VFA was about 34% higher in faunated than in non-faunated calves (P < 0.001) . Hydrogen (2H) recovery of rumen fermentation was positively correlated (R2 = 0.55) to the CH4 production rate . Methanogens were attached on rumen mucosa . Methanogenesis, induced by rumen mucosa attached bacteria, was stimulated by ruminal protozoa . In the absence of protozoa in the rumen, the acetate-propionate ratio and butyrate proportion of VFA were reduced . In vivo, in the absence of protozoa not only the whole animal CH4 production (-30%, P < 0.05) but also the digestibility of carbohydrates (-4%, P < 0.05) was reduced . Thereby no difference was observed in the intake of ME per kg DM between the groups . In conclusion, the methanogenesis in the rumen, but not in hindgut, is associated with the development of the ruminal protozoa population . The level of methanogenesis (mol/mol VFA) in the hindgut amounts to 20% of the ruminal methanogenesis. Antonie Van Leeuwenhoek, 2003, 84(2), 135 - 45 Exopolysaccharide production by filamentous fungi: the example of Botryosphaeria rhodina; Selbmann L et al.; One-hundred and five fungal strains, belonging to 46 different species, were screened for exopolysaccharide production . Phytopathogenicity and, in particular, inability to produce conidia, were physiological characteristics positively associated and correlated with the fungal ability to produce polysaccharides . Among the 29 positive strains, Botryosphaeria rhodina DABAC-P82 was the most interesting reaching, when grown on optimal nitrogen source and concentration (NaNO3 and 2.0 g l(-1), respectively) and culture medium pH (3.7), 17.7 g l(-1) of exopolysaccharide production after only 24 h of fermentation; yield and productivity were 0.69 g g(-1) and 0.73 g l(-1)h(-1), respectively . The purified polysaccharide was characterised as a homopolysaccharide of glucose with a molecular weight of 4.875 x 10(6) Da . Studies of structural analysis indicated the presence of beta-1,3 and beta-1,6 linkages; the EPS structure was very similar to that of scleroglucan. Appl Environ Microbiol, 2003 Oct, 69(10), 6091 - 8 Influence of nutritional factors on the nature, yield, and composition of exopolysaccharides produced by Gluconacetobacter xylinus I-2281; Kornmann H et al.; The influence of substrate composition on the yield, nature, and composition of exopolysaccharides (EPS) produced by the food-grade strain Gluconacetobacter xylinus I-2281 was investigated during controlled cultivations on mixed substrates containing acetate and either glucose, sucrose, or fructose . Enzymatic activity analysis and acid hydrolysis revealed that two EPS, gluconacetan and levan, were produced by G . xylinus . In contrast to other acetic acid strains, no exocellulose formation has been measured . Considerable differences in metabolite yields have been observed with regard to the carbohydrate source . It was shown that glucose was inadequate for EPS production since most of this substrate (0.84 C-mol/C-mol) was oxidized into gluconic acid, 2-ketogluconic acid, and 5-ketogluconic acid . In contrast, sucrose and fructose supported a 0.35 C-mol/C-mol gluconacetan yield . In addition, growing G . xylinus on sucrose produced a 0.07 C-mol/C-mol levan yield . The composition of EPS remained unchanged during the course of the fermentations . Levan sucrase activity was found to be mainly membrane associated . In addition to levan production, an analysis of levan sucrase's activity also explained the formation of glucose oxides during fermentation on sucrose through the release of glucose . The biosynthetic pathway of gluconacetan synthesis has also been explored . Although the activity of key enzymes showed large differences to be a function of the carbon source, the ratio of their activities remained similar from one carbon source to another and corresponded to the ratio of precursor needs as deduced from the gluconacetan composition. Appl Environ Microbiol, 2003 Oct, 69(10), 5892 - 7 Engineering redox cofactor regeneration for improved pentose fermentation in Saccharomyces cerevisiae; Verho R et al.; Pentose fermentation to ethanol with recombinant Saccharomyces cerevisiae is slow and has a low yield . A likely reason for this is that the catabolism of the pentoses D-xylose and L-arabinose through the corresponding fungal pathways creates an imbalance of redox cofactors . The process, although redox neutral, requires NADPH and NAD+, which have to be regenerated in separate processes . NADPH is normally generated through the oxidative part of the pentose phosphate pathway by the action of glucose-6-phosphate dehydrogenase (ZWF1) . To facilitate NADPH regeneration, we expressed the recently discovered gene GDP1, which codes for a fungal NADP+-dependent D-glyceraldehyde-3-phosphate dehydrogenase (NADP-GAPDH) (EC 1.2.1.13), in an S . cerevisiae strain with the D-xylose pathway . NADPH regeneration through an NADP-GAPDH is not linked to CO2 production . The resulting strain fermented D-xylose to ethanol with a higher rate and yield than the corresponding strain without GDP1; i.e., the levels of the unwanted side products xylitol and CO2 were lowered . The oxidative part of the pentose phosphate pathway is the main natural path for NADPH regeneration . However, use of this pathway causes wasteful CO2 production and creates a redox imbalance on the path of anaerobic pentose fermentation to ethanol because it does not regenerate NAD+ . The deletion of the gene ZWF1 (which codes for glucose-6-phosphate dehydrogenase), in combination with overexpression of GDP1 further stimulated D-xylose fermentation with respect to rate and yield . Through genetic engineering of the redox reactions, the yeast strain was converted from a strain that produced mainly xylitol and CO2 from D-xylose to a strain that produced mainly ethanol under anaerobic conditions. Water Sci Technol, 2003, 48(4), 249 - 52 Start-up of full-scale anaerobic digesters treating municipal solid waste; Ohmura T et al.; Raising organic loading rate, and the behavior of dissolved CODcr (D-CODcr), VFA and specific methanogen activity, were investigated through a laboratory experiment for the start-up of a sludge recycling center . Moreover, application for MPN-PCR methods using a gene as a direct technique to measure the quantity of methanogen was attempted . It was recognized that specific methanogen activity depends on the quantity of methanogen, and that gas production does not reflect the condition of methane fermentation . The methane fermentation condition was checked through the specific methanogen activity and analysis of D-CODcr . The target organic loading rate was reached in the short period of about 30 days, and rapid start-up was successfully attained for a full-scale anaerobic digester. Water Sci Technol, 2003, 48(4), 213 - 6 Degradation of plant wastes by anaerobic process using rumen bacteria; Seon J et al.; An operational reactor has been designed for the fermentation of a pure culture of Fibrobacter succinogenes with the constraints of strict anaerobic condition . The process is controlled by measurements of pH, redox, temperature and CO2 pressure; it allows an efficient degradation (67%) of lignocellulosic wastes such as a mixture of wheat straw, soya bean cake and green cabbage. Water Sci Technol, 2003, 48(4), 195 - 8 Organic material solubilization of domestic primary sludge in anaerobic digestion at controlled pH; Gomec CY et al.; The role of the pH in the anaerobic digestion of domestic primary sludge at mesophilic temperature was evaluated . Since solubilization of organic materials is seriously affected by pH, much research has been performed for the evaluation of significant pH influence on the production of soluble organic carbon in the acid phase . In this study, the performance of continuously stirred anaerobic reactors for the production of soluble chemical oxygen demand (SCOD) and volatile fatty acids (VFAs) in domestic primary sludge was investigated . In the first anaerobic reactor, the pH was not controlled (pH-uncontrolled) whereas the pH was fixed at 6.5 (pH-controlled) in the second reactor . Two sets were performed . Results of both sets indicated that, VSS was removed with a corresponding production of VFAs and SCOD in both reactors . However, production of VFAs and SCOD stopped earlier in the pH-controlled reactors . Methanogenic phase started to operate at earlier times whereas hydrolysis and fermentation were still operating in the pH-uncontrolled reactors . The process stability and efficiency of anaerobic digestion of substrates such as domestic primary sludge can be improved by anaerobic digestion elutriated phased treatment (ADEPT) in which the acid elutriation reactor and methanogenic reactor are separated . In this respect, ADEPT was introduced. Water Sci Technol, 2003, 48(4), 77 - 84 Alkaline subcritical-water treatment and alkaline heat treatment for the increase in biodegradability of newsprint waste; Fox MH et al.; This work describes two alkaline semicontinuous processes for the conversion of refractory organic materials into biodegradable substances . Newsprint was used as a lignocellulosic waste . Methane conversion efficiencies and cellulose removals were investigated for the two following processes: alkaline subcritical-water treatment (ASWT) coupled with methane fermentation and alkaline heat treatment (newsprint heated with steam in an autoclave; AHT) coupled with methane fermentation with a neutral subcritical-water treatment (NSWT) recycle . Results showed that for ASWT coupled with methane fermentation higher methane conversion efficiencies and higher cellulose removals were achieved as HRT increased . At HRT = 20 days, average CH4 conversion efficiency and average cellulose removal reached 26% and 44%, respectively . After a final HRT of 40 days, average CH4 conversion efficiency and average cellulose removal reached 50% and 60%, respectively . On the other hand, for AHT coupled with methane fermentation, methane conversion efficiencies did not show a greater improvement using this pretreatment process . Average conversion reached 9% with an average cellulose removal of 20% . In order to improve the yield of the reactor, approximately one-third of the effluent was recycled using NSWT (150 degrees C; neutral pH) . Methane conversion efficiency of this process increased as more recycles were performed . For the fifth operation, the total average methane conversion efficiency was 44% with a total average cellulose removal of 55%. Biofactors, 2002, 16(3-4), 105 - 15 Amelioration of cisplatin toxicity by a fermented grain food product; Minamiyama Y et al.; The most noticeable hypothesis regarding the pathogenesis of cisplatin toxicity, seen mainly in kidney and intestine, is oxidative stress, an imbalance between free-radical generating cisplatin and radical scavenging systems . This paper describes the role of the antioxidant system in cisplatin-induced toxicity and the protective effect by a processed grain food (Antioxidant Biofactor: AOB), which has been shown to exhibit strong antioxidant activity . Male Fischer 344 rats were used . They were pre-fed either a basal diet (control, 15 g/day) or the diet supplemented with AOB to provide 6.5% or 20% of total diet throughout the experiment . Cisplatin (5 mg/kg, i.v.) was administered at the start of the experiment, and the animals were sacrificed 5 days later . Blood urea nitrogen (BUN) and plasma creatinine, NO2(-) and NO3(-) (NOx) were determined from the plasma . The levels of 4-hydroxy-2-nonenal (a lipid peroxidation product), 8-hydroxy-deoxyguanosine (8-OHdG, an oxidatively modified DNA adduct) and nitrotyrosine were histologically analyzed . The cisplatin administration resulted in a loss of body weight and elevations of BUN, serum creatinine and NOx levels, whereas AOB supplement reversed these effects . The severe morphological damages induced in the kidney and intestine by the cisplatin administration were markedly improved in the AOB group . The levels of lipid peroxidation, 8-OHdG, and nitrotyrosine all paralleled the morphological damage . The AOB effect was dose dependent . In conclusion, the present study suggests that certain food additives like AOB may be of benefit against the side effects of cisplatin. Syst Appl Microbiol, 2003 Sep, 26(3), 471 - 9 Arginase activity is a useful marker of nitrogen limitation during alcoholic fermentations; Carrasco P et al.; Nitrogen deficiency in musts is one of the causes of sluggish or stuck fermentations . In this work we propose that arginase activity determination can be useful for detecting nitrogen starvation early in vinification . CAR1 and YGP1 genes are not specifically induced under conditions of nitrogen starvation . However, a significant increase in the enzymatic activity of arginase, the product of the CAR1 gene, is detected in vinifications carried out with musts containing limiting amounts of nitrogen . Moreover, on adding ammonia to a nitrogen-deficient vinification, even at late stages, this enzymatic activity is repressed, and growth rate is restored simultaneously . We also investigate the role of ethanol toxicity in nitrogen starvation . The results suggest that ethanol produced during vinification or exogenously added up to 8% (v/v) concentration does not cause nitrogen starvation under the conditions tested because arginase activity is not increased. J Anat, 2003 Sep, 203(3), 277 - 82 Age-associated plasticity in the intrinsic innervation of the ovine rumen; Pfannkuche H et al.; The rumen of adult sheep functions as a large fermentation chamber . In the newborn suckling ruminant, the rumen is bypassed and milk enters the abomasum directly . It was the aim of our study to investigate whether the transmitter content of intrinsic nerves changes with the developmental stage . The neurochemical code of myenteric neurons in the rumen from suckling lambs, fattened lambs and adult sheep was determined by using quadruple immunohistochemistry against choline-acetyltransferase (ChAT), nitric oxide synthase (NOS), substance P (SP) and vasoactive intestinal peptide (VIP) . Three neurochemically distinct subpopulations were identified within the rumen . They expressed the code ChAT/-, ChAT/SP and NOS/VIP . The number of ChAT/SP neurons did not change during development . It was 62% in the newborn lamb and remained stable in fattened lambs (63%) and adult sheep (63%) . By contrast, the number of ChAT/- neurons decreased significantly from 20% in suckling lambs to 11% and 7% in fattened lambs and adult sheep, respectively . Simultaneously, the proportion of NOS/VIP neurons increased from 16% in suckling lambs to 29% in adult sheep . The increase in the proportion of NOS/VIP immunoreactive neurons indicates an adaptation to large volumes of ingesta at the beginning of roughage intake and rumination . We conclude that the age-associated changes in neurochemical code of myenteric neurons in the forestomach are related to the adaption of the rumen to different functional properties during development. Int J Food Microbiol, 2003 Nov 15, 88(1), 41 - 54 Biogenic amines in dry fermented sausages: a review; Suzzi G et al.; Biogenic amines are compounds commonly present in living organisms in which they are responsible for many essential functions . They can be naturally present in many foods such as fruits and vegetables, meat, fish, chocolate and milk, but they can also be produced in high amounts by microorganisms through the activity of amino acid decarboxylases . Excessive consumption of these amines can be of health concern because their not equilibrate assumption in human organism can generate different degrees of diseases determined by their action on nervous, gastric and intestinal systems and blood pressure . High microbial counts, which characterise fermented foods, often unavoidably lead to considerable accumulation of biogenic amines, especially tyramine, 2-phenylethylamine, tryptamine, cadaverine, putrescine and histamine . However, great fluctuations of amine content are reported in the same type of product . These differences depend on many variables: the quali-quantitative composition of microbial microflora, the chemico-physical variables, the hygienic procedure adopted during production, and the availability of precursors . Dry fermented sausages are worldwide diffused fermented meat products that can be a source of biogenic amines . Even in the absence of specific rules and regulations regarding the presence of these compounds in sausages and other fermented products, an increasing attention is given to biogenic amines, especially in relation to the higher number of consumers with enhanced sensitivity to biogenic amines determined by the inhibition of the action of amino oxidases, the enzymes involved in the detoxification of these substances . The aim of this paper is to give an overview on the presence of these compounds in dry fermented sausages and to discuss the most important factors influencing their accumulation . These include process and implicit factors as well as the role of starter and nonstarter microflora growing in the different steps of sausage production . Moreover, the role of microorganisms with amino oxidase activity as starter cultures to control or reduce the accumulation of biogenic amines during ripening and storage of sausages is discussed. Biotechnol Prog, 2003 Sep-Oct, 19(5), 1615 - 9 Novel fermentation strategy for enhancing glycerol production by Candida krusei; Liu HJ et al.; During the later stage of glycerol production by fermentation of Candida krusei, glycerol consumption by the strain was observed, although there was residual sugar in the medium . To enhance the final glycerol accumulation, a new fermentation strategy was developed by maintaining high activities of glycerol synthetic enzymes (i.e., glycerol-3-phosphate dehydrogenase (ctGPD) and glycerol-3-phosphatase (GPP)) for a relatively long period while conducting oxygen limitation at a later stage to inhibit the increase of another enzyme activity related to glycerol degradation (i.e., mitochondrial glycerol-3-phosphate dehydrogenase (mtGPD)) . With oxygen limitation performed from 88 h, when ctGPD and GPP activities were already at a low level while mtGPD activity was increasing, the glycerol dissimilation was efficiently reduced . The final glycerol concentration reached 55.6 g/L, which was about 18% (96 h) and 30% (104 h) higher than control, and its productivity increased to 0.54 g/(L h) . The proposed strategy based on cell physiology was proved useful to the glycerol fermentation process. Biotechnol Prog, 2003 Sep-Oct, 19(5), 1606 - 11 Effect of oxygen delignification operating parameters on downstream enzymatic hydrolysis of softwood substrates; Charles N et al.; Enhanced oxygen delignification of softwood pulp samples (taken upstream and downstream of a commercial oxygen delignification unit) improved the initial rate of enzymatic saccharification and overall yield of monomeric sugars by 62-82% and 76-80%, respectively . Laboratory-scale experiments were used to examine the effect of a broad range of operating parameters (temperature, time, caustic concentration, and oxygen partial pressure) on the effectiveness of oxygen delignification . Using empirical models, kappa number (residual lignin content) was found to effectively predict final conversion to monomeric sugars . Application of oxygen delignification to sulfite mill knots resulted in smaller (20-25%) reduction in lignin content . However, using a combination of oxygen delignification and particle size reduction, up to 80% of the carbohydrate in the reject knots could be converted to fermentable sugars. Biotechnol Prog, 2003 Sep-Oct, 19(5), 1591 - 605 Data-based modeling and analysis of bioprocesses: some real experiences; Karim MN et al.; Data-generated models find numerous applications in areas where the speed of collection and logging of data surpasses the ability to analyze it . This work is meant to addresses some of the challenges and difficulties encountered in the practical application of these methods in an industrial setting and, more specifically, in the bioprocess industry . Neural network and principal component models are the two topics that are covered in detail in this paper . A review of these modeling technologies as applied to bioprocessing is provided, and four original case studies using industrial fermentation data are presented that utilize these models in the context of prediction and monitoring of bioprocess performance. Biotechnol Prog, 2003 Sep-Oct, 19(5), 1468 - 79 Study of process variables in industrial acetic fermentation by a continuous pilot fermentor and response surfaces; Garrido-Vidal D et al.; The modeling and optimization of industrial processes requires an intensive study of the factors involved . In this work, a continuous pilot system for studying the industrial process of acetic fermentation is developed . A Doehlert design is applied to the five variables involved in the pilot process . This experimental design allows reduction of the experimental burden and the maximum amount of information to be obtained, studying the factors at different levels depending on their significance . The experimental system provides a robust measure of the specific growth rate and the rates of substrates consumption and acetic acid production, related to the flow of effluent stream evaluated in the steady state . The results demonstrate the growth-associated kinetics of substrates and product, and the yield factors are calculated with low values of variances for the coefficients, i.e., within the range 1-11% . The specific growth rate suits the quadratic model proposed . The response surfaces generated by the model are applied to explain the behavior of the bacterial growth and, therefore, the effects of the process variables studied over the acetic acid production . Very low levels of ethanol or oxygen make the acetification rate decrease, and a saturation effect with high levels of ethanol or oxygen is also deduced . The effects of the aeration rate, agitation, and overpressure suggest a kind of inhibition of the acetic acid production caused by the oxygen that has not been practically studied before . The temperature strengthens the inhibitory effect of the ethanol and the oxygen . The conclusions of this work consolidate the structure of a hybrid model for the acetic fermentation. Microbiology, 2003 Oct, 149(Pt 10), 2873 - 8 Aggregation of yeast cells: direct measurement of discrete lectin-carbohydrate interactions; Touhami A et al.; Aggregation of microbial cells mediated by specific interactions plays a pivotal role in the natural environment, in medicine and in biotechnological processes . Here we used atomic force microscopy (AFM) to measure individual lectin-carbohydrate interactions involved in the flocculation of yeast cells, an aggregation event of crucial importance in fermentation technology . AFM probes functionalized with oligoglucose carbohydrates were used to record force-distance curves on living yeast cells at a rate of 0.5 micro m s(-1) . Flocculating cells showed adhesion forces of 121+/-53 pN, reflecting the specific interaction between individual cell-surface lectins and glucose residues . Similar adhesion forces, 117+/-41 pN, were measured using probes functionalized with the lectin concanavalin A and attributed to specific binding to cell-surface mannose residues . By contrast, specific interaction forces were not observed in non-flocculating conditions, i.e . in the presence of mannose or when using non-flocculating cells, pointing to their involvement in yeast flocculation . The single molecule force spectroscopy measurements presented here provide a means to study a variety of cellular interactions at the molecular level, such as the adhesion of bacteria to animal and plant tissues. Am J Clin Nutr, 2003 Oct, 78(4), 881S - 892S Sugars and dental caries; Touger-Decker R et al.; A dynamic relation exists between sugars and oral health . Diet affects the integrity of the teeth; quantity, pH, and composition of the saliva; and plaque pH . Sugars and other fermentable carbohydrates, after being hydrolyzed by salivary amylase, provide substrate for the actions of oral bacteria, which in turn lower plaque and salivary pH . The resultant action is the beginning of tooth demineralization . Consumed sugars are naturally occurring or are added . Many factors in addition to sugars affect the caries process, including the form of food or fluid, the duration of exposure, nutrient composition, sequence of eating, salivary flow, presence of buffers, and oral hygiene . Studies have confirmed the direct relation between intake of dietary sugars and dental caries across the life span . Since the introduction of fluoride, the incidence of caries worldwide has decreased, despite increases in sugars consumption . Other dietary factors (eg, the presence of buffers in dairy products; the use of sugarless chewing gum, particularly gum containing xylitol; and the consumption of sugars as part of meals rather than between meals) may reduce the risk of caries . The primary public health measures for reducing caries risk, from a nutrition perspective, are the consumption of a balanced diet and adherence to dietary guidelines and the dietary reference intakes; from a dental perspective, the primary public health measures are the use of topical fluorides and consumption of fluoridated water. J Nutr, 2003 Oct, 133(10), 3141 - 4 Fermentable and nonfermentable fiber supplements did not alter hunger, satiety or body weight in a pilot study of men and women consuming self-selected diets; Howarth NC et al.; Little is known about the relative effects of fermentable fiber (FF) vs . nonfermentable fiber (NFF) on energy regulation in humans . We compared 27 +/- 0.6 g/d supplements of FF (pectin, beta-glucan) and NFF (methylcellulose) for their ability to decrease ad libitum energy intake (EI) and hunger, increase satiety and cause spontaneous body weight and fat losses . Men and women (n = 11) aged 23-46 y, BMI 20.0-34.4 kg/m2, consumed first NFF and then FF for 3 wk each, with a 4-wk washout period between phases . Daily satiety assessed with analog scales was higher with NFF than FF (60.7 +/- 1.0 vs . 57.7 +/- 0.8 mm, P = 0.01) . However, there were no differences in reported EI (NFF < FF by 7%, P = 0.31, NFF < baseline by 9.5%, P = 0.11), body weight (NFF 0.13 kg, P = 0.73; FF 0.13 kg, P = 0.60) or fat percentage (NFF -0.3%, P = 0.56; FF -0.1%, P = 0.66) within either phase . In contrast to findings in animals, NFF was more, rather than less satiating than FF, and use of neither NFF nor FF preparations was associated with body weight or fat loss . These pilot results suggest no role for short-term use of FF and NFF supplements in promoting weight loss in humans consuming a diet ad libitum. J Nutr, 2003 Oct, 133(10), 3080 - 4 A pilot study to investigate effects of inulin on Caco-2 cells through in vitro metabolic fingerprinting; Lamers RJ et al.; Metabolic fingerprints are novel measurement tools to evaluate the biochemical status of a living organism by using 1H NMR and multivariate data analysis (MVDA) . In this way, a quick evaluation of changes in health or diseased state can be made, reflected in alterations of metabolic patterns . Normally, metabolic fingerprinting is based on in vivo studies . These studies often represent a labor-intensive and expensive manner of investigation . In vitro studies are not hampered by these disadvantages, thus constituting an interesting alternative . In this research, results are presented of a pilot experiment in which metabolic fingerprinting was combined with an in vitro model . For this purpose, differentiated Caco-2 cells were exposed to inulin and its fermentative metabolites, both dissolved in culture medium . Cells were incubated for 0 or 48 h . Cell fractions were analyzed by NMR, then subsequently with MVDA . Differences in treatment provided detectable variations in the time of metabolic patterns of cell contents . Results indicated that glucose metabolism linked to glutamate was of major importance in the effects of inulin and its metabolites on Caco-2 cells under the conditions of our study . Metabolic fingerprinting in combination with an in vitro model appears to be a feasible technique with which to visualize metabolic patterns of cell contents and provides an efficient place for the generation of hypotheses about the metabolic pathways involved . In vitro metabolic fingerprinting may be of great benefit in the future for a better understanding of the relationship between nutrition and health. Vet Parasitol, 2003 Oct 8, 116(2), 125 - 38 Effects of Oesophagostomum dentatum and dietary carbohydrates on morphology of the large intestine of pigs; Petkevicius S et al.; The effects of Oesophagostomum dentatum infection and dietary carbohydrates on the morphology and epithelial cell proliferation in the gastrointestinal tract of pigs were investigated experimentally . Thirty-two worm-free pigs (n=32) from a specific pathogen-free farm were randomly divided into four groups (A-D), of eight animals each . Pigs in groups A (control) and B (infected) were fed Diet 1, and pigs in groups C (control) and D (infected) were fed Diet 2 . The two diets were formulated: Diet 1 (%) contained barley flour, oat husk meal plus soya bean meal (55:21:24) and Diet 2 (%) contained barley flour, inulin and sugar beet fibre (SBF) (80.1:7:12.9) plus soya bean meal (3:1) to contain carbohydrates from inulin and sugar beet fibre (SBF) that were readily fermentable in the large intestine . The two infected pig groups (16 pigs total) were inoculated with 6000 infective larvae of O . dentatum and all pigs, including the controls, were slaughtered 12 weeks p.i . The combination of O . dentatum infection and highly fermentable dietary carbohydrates affected the mucosal architecture, the epithelial cell proliferation and mucin secretion of the large intestine . Infection had a significant influence on the crypt volume, height and density, and on muscularis externa at the proximal and middle colon . The changes in the affected gut sections were proportional to the number of worms present . However, these parameters appeared unaffected by those diets alone . In pigs without infection non-digestible dietary carbohydrates significantly influenced the tissue weight of colon. J Agric Food Chem, 2003 Oct 8, 51(21), 6191 - 7 Leavened dough processing by supercritical fluid extrusion (SCFX); Hicsasmaz Z et al.; Yeast-leavened dough processing is semicontinuous due to the requirement for fermentation at constant temperature and humidity . Also, new regulations on the emission of alcohols are becoming burdensome on the baking industry . Extrusion processing of dough with supercritical carbon dioxide (SC-CO(2)) is envisioned to alleviate emission problems and to decrease production time by eliminating fermentation . A bread dough formulation with 50% (w/w) moisture was leavened by injecting 1.5% (w/w) SC-CO(2) in a twin-screw extruder at 37 degrees C . Specific mechanical energy input was 260 kJ/kg . The operating apparent shear rate range was 60-260 s(-1) . SCFX-leavened dough density (420-430 kg/m(3)) was in good agreement with values reported for similar doughs . The flow behavior index, obtained by an on-line slit rheometer, was 0.49 for the nonleavened control and 0.63 for the SCFX-leavened dough . Apparent viscosity of the SCFX-leavened dough varied from 37 to 23 Pa-s . This new continuous process offers attractive possibilities for industrial applications if further developed. Can Bull Med Hist, 2001, 18(2), 297 - 323 "Stand up and be counted": nursing at the Calgary General Hospital after the Second World War; Richardson SL; The post-second World War period of reconstruction was a time of ferment for Canadian hospitals, and consequently, for hospital nursing work and education . Demand for hospital services, especially nursing, increased dramatically . At the same time, fewer young women were willing to enroll in hospital nurse-training programs . This article is a case study of how one voluntary western hospital - the Calgary General Hospital - grappled with the post-second World War shortage of both graduate nurses and student nurses, at a time of transition from a small, cottage-type hospital into a large, complex urban institution . The analysis offers insights into today's economically driven and politically controlled health care delivery system restructuring, while illuminating the contributions of one of Canada's least known nurse leaders - Gertrude May Hall. J Antibiot (Tokyo), 2003 Jul, 56(7), 617 - 21 Gerfelin, a novel inhibitor of geranylgeranyl diphosphate synthase from Beauveria felina QN22047 . I . Taxonomy, fermentation, isolation, and biological activities; Zenitani S et al.; A new compound, gerfelin, was isolated from a culture broth of Beauveria felina QN22047 . It was purified by column chromatography on silica gel and by HPLC . Gerfelin has the molecular formula C15H14O6 . It inhibited synthesis of geranylgeranyl diphosphate, which was mediated by recombinant human geranylgeranyl diphosphate synthase (hGGPP synthase) in vitro . The inhibitory pattern of gerfelin was noncompetitive against isopentenyl diphosphate, and uncompetitive against farnesyl diphosphate. J Antibiot (Tokyo), 2003 Jul, 56(7), 603 - 9 EI-1941-1 and -2, novel interleukin-1 beta converting enzyme inhibitors Produced by Farrowia sp . E-1941 . II . Taxonomy of producing strain, fermentation, isolation, physico-chemical properties, and biological properties; Koizumi F et al.; EI-1941-1 and -2, novel interleukin-1 beta converting enzyme (ICE) inhibitors, were isolated from the culture broths of Farrowia sp . E-1941 . EI-1941-1 and -2 selectively inhibited the human recombinant ICE activity with IC50 values of 0.086 and 0.006 microM, respectively . Taxonomy and fermentation of the producing strain and isolation, physico-chemical properties, structure elucidation, and biological properties of EI-1941-1 and -2 are described. J Ind Microbiol Biotechnol, 2003 Sep, 30(9), 568 - 76 Epub 2003 Aug 28. Cloning and expression of fungal phytases in genetically modified strains of Aspergillus awamori; Martin JA et al.; In an effort to produce phytases cost-effectively, and to determine the efficiency of a novel expression system, the genes for Aspergillus awamori ( phyA) phytase and Aspergillus fumigatus ( phyA) phytase (a putative thermostable enzyme) were cloned and overexpressed in A . awamori . Regulation of phytase expression was achieved by separately placing the genes under the transcriptional control of the glucoamylase A ( glaA) promoter of A . awamori . A gene fusion strategy was employed that involved the insertion of a hexapeptide Kex-2 protease cleavage site between the native glucoamylase and heterologous proteins and allowed for the efficient secretion and processing of the resultant chimeric proteins produced in this system by an endogenous Kex-2 protease . The genes for both of the above-mentioned phytases have already been cloned; however, this is the first report of either of the two phytases being fused with the glucoamylase gene, placed under the transcriptional control of the glaA promoter and overexpressed in A . awamori . Following transformation of A . awamori with separate expression vectors (one for each phytase), induction of phytase expression in submerged culture was effected by utilisation of a starch-containing medium . Optimisation of heterologous protein production in small shake-flask cultures involved changes in medium constituents . Maximum phytase expression levels of 200 phytase units (PU) ml(-1) and 62 PU ml(-1) for recombinantly expressed phytases from A . awamori and A . fumigatus, respectively, were obtained in crude fermentation extracts . Subsequent process scale-up to 4 l batch fermentation yielded phytase production levels comparable to those obtained on small scale . The enzyme yields herein reported demonstrate that the expression system developed and the host strain utilised were capable of expressing phytase at levels comparable to, or exceeding, previously reported data. Biochem Biophys Res Commun, 2003 Oct 10, 310(1), 196 - 201 Overexpression and purification of the vanilloid receptor in yeast (Saccharomyces cerevisiae); Moiseenkova VY et al.; The vanilloid receptor type 1 (VR1) is a novel membrane receptor activated by heat or chemical ligands conveying information about chemosensitive and thermosensitive pain . We have overexpressed and purified wild type VR1 (wtVR1) as well as several mutant forms using the yeast strain Saccharomyces cerevisiae with the goal of obtaining sufficient protein for structural studies . To facilitate the rapid assaying of protein production and purification we used PCR to construct mutant VR1-green fluorescent protein fusion genes . All recombinant inserts were engineered with 12 HIS tags on the C-terminus for metal affinity column purification . The yield of purified protein from 16L fermentation was about 1mg following a single-step purification procedure . By taking advantage of the calcium permeability of VR1 we measured changes in {Ca(2+)}(i) in capsaicin-stimulated fura-2 loaded yeast cells expressing VR1. J Anim Physiol Anim Nutr (Berl), 2003 Apr, 87(3-4), 116 - 21 Effects of different feeding and housing conditions on dry matter content and consistency of faeces in sows; Tabeling R et al.; In a study with sows at various stages of the reproductive cycle, the effects of parturition, feed quantity and composition (content and type of crude fibre), exercise and access to water on dry matter content, and consistency of faeces were estimated . Parturition caused an increase in the dry matter content and reduced the defaecation frequency . The restriction of feed amounts in the last days of gestation and the change to lactation feed with reduced crude fibre content (6-3% in feed) intensified the physiologically occurring increase of the dry matter content . Moving activity and the amount of water ingested had only a small insignificant effect on the moisture of the faeces . The faeces became markedly softer when large amounts of fermentable fibre were used in the diet . The risk for constipation in periparturient sows increases when feed amounts and fibre content in the diet are strongly reduced . Therefore, it is recommended to offer sows daily minimum amounts of a diet (<2 kg) containing >7-8% crude fibre (including a highly fermentable fibre to improve faeces consistency) especially on the days near parturition. Bioprocess Biosyst Eng, 2002 Sep, 25(3), 183 - 91 Epub 2002 Aug 09. On-line estimation of sugar concentration for control of fed-batch fermentation of lignocellulosic hydrolyzates by Saccharomyces cerevisiae; Nilsson A et al.; A feed control strategy, based on estimated sugar concentrations, was developed with the purpose of avoiding severe inhibition of the yeast Saccharomyces cerevisiae during fermentation of spruce hydrolyzate . The sum of the fermentable hexose sugars, glucose and mannose, was estimated from on-line measurements of carbon dioxide evolution rate and biomass concentration by use of a simple stoichiometric model . The feed rate of the hydrolyzate was controlled to maintain constant sugar concentration during fed-batch fermentation, and the effect of different set-point concentrations was investigated using both untreated and detoxified hydrolyzates . The fed-batch cultivations were evaluated with respect to cellular physiology in terms of the specific ethanol productivities, ethanol yields, and viability of the yeast . The simple stoichiometric model used resulted in a good agreement between estimated sugar concentrations and off-line determinations of sugar concentrations . Furthermore, the control strategy used made it possible to maintain a constant sugar concentration without major oscillations in the feed rate or the sugar concentration . For untreated hydrolyzates the average ethanol productivity could be increased by more than 130% compared to batch fermentation . The average ethanol productivity was increased from 0.12 to 0.28 g/g h . The productivity also increased for detoxified hydrolyzates, where an increase of 16% was found (from 0.50 to 0.58 g/g h). Bioprocess Biosyst Eng, 2002 Sep, 25(3), 179 - 81 Epub 2002 Jul 27. Comparison of fumaric acid production by Rhizopus oryzae using different neutralizing agents; Zhou Y et al.; Fumaric acid fermentation in a 10-L bubble column fermenter using different neutralizing agents {CaCO(3), Ca(OH)(2), NaHCO(3)} by Rhizopus oryzae ATCC 20344 was examined . It was found that in the fermentation using CaCO(3 )as the neutralizing agent the highest fumaric acid weight yield and volumetric productivity were obtained, 53.4% and 1.03 g/L x h(-1) respectively . In the NaHCO(3) case, the fumaric acid weight yield and volumetric productivity were 33.7% and 0.69 g/L x h(-1), respectively, much lower than the CaCO(3) case . However, the NaHCO(3) alternative has advantages of cell reuse and simple downstream processing because of the high solubility of sodium fumarate . These advantages may offset the disadvantages of using NaHCO(3) as the neutralizing agent, and the overall fumaric acid weight yield and volumetric productivity will increase. Bioprocess Biosyst Eng, 2002 Sep, 25(3), 165 - 8 Epub 2002 Jun 13. A combined solid-state and submerged cultivation integrated with adsorptive product extraction for production of Monascus red pigments; Hsu FL et al.; A process taking advantages of combined solid-state and submerged cultivation of Monascus for red pigment production and integration of a product removal unit was developed . The solid-state cultivation was carried out in a 5 l fermentor, with rice being used not only as the substrate but also the support for Monascus . The inclusion of rice submergence and integration of product separation were achieved by intermittently rinsing the rice with monosodium glutamate (MSG) solutions every 12 h followed by an adsorptive extraction of the red pigment dissolved in the rinsing solution . With this new process, the Monascus red pigment production was increased by 24% as compared with that by the plain fixed-bed cultivation. J Dairy Sci, 2003 Sep, 86(9), 2949 - 57 Influence of corn silage particle length on the performance of lactating dairy cows fed supplemental tallow; Onetti SG et al.; The objective of this study was to determine if the length of chop of processed corn silage influences the impact of supplemental fat on rumen fermentation and performance of dairy cows . We hypothesized that increasing forage particle length may alleviate the interference of fat on rumen fermentation . Sixteen Holstein cows averaging 120 d in milk were used in a replicated 4 x 4 Latin square design with 21-d periods . Treatments were arranged as a 2 x 2 factorial with 0 or 2% tallow (dry matter basis), and corn silage harvested at either 19 or 32 mm theoretical length of cut . The forage:concentrate ratio was 50:50, and diets were formulated to contain 18% crude protein and 32% neutral detergent fiber (dry matter basis) . Cows were allowed ad libitum consumption of diets that were fed twice daily as a total mixed ration . Fat supplemented cows had lower dry matter intake and produced less milk fat relative to nonsupplemented cows . No effect of corn silage particle length was observed for dry matter intake and milk fat production . Proportion of trans-10 C18:1 and of trans-10, cis-12 conjugated linoleic acid was highest in milk fat of cows fed 2% supplemental tallow . Rumen pH was not affected by feeding tallow, and tended to be highest for cows eating the 32-mm theoretical length of chop corn silage diets . No effect of treatments was observed for rumen acetate-to-propionate ratio or rumen ammonia concentration . In this study, tallow supplementation had a negative impact on performance of dairy cows regardless of the corn silage particle length . Feeding tallow increased formation of trans-fatty acids in the rumen in the absence of significant changes in the rumen environment. Bioprocess Biosyst Eng, 2003 Mar, 25(5), 309 - 14 Epub 2003 Feb 08. Filtration of a bacterial fermentation broth: harvest conditions effects on cake hydraulic resistance; Meireles M et al.; The hydraulic resistance of cakes formed during the ultrafiltration of Streptomyces pristinaespiralis broths has been investigated for different harvesting conditions . S . pristinaespiralis broth was harvested after the point of microorganism activity declines (0-h aged broth) and afterwards held for different durations of up to 16 h (16 aged broths) . Aging behavior occurring between the end of microorganism activity and harvest was compared for different acidification procedures (pH) and the mechanisms for which the hydraulic resistance of the cake is affected by aging have been investigated . For broths harvested under conditions where the acidification is fixed at pH 2 or 3, hydraulic resistance associated with cake build-up is directly determined by the interactions between the cells . Holding broths beyond 5 h contributes to a release of a soluble component from the cell surface . Enhanced cell surface interactions then turn the cake structure into a more open one and reduce the specific hydraulic resistance . For broths harvested under conditions where the acidification is fixed at pH 4, hydraulic resistance associated with cake build-up is both determined by cell interactions and cell morphology . The cause of the increase in specific hydraulic resistance with aging is due to the binding of a soluble component released by the microorganisms, which decreases the cell surface interactions. Bioprocess Biosyst Eng, 2003 Mar, 25(5), 263 - 70 Epub 2003 Feb 06. Functional nodes in dynamic neural networks for bioprocess modelling; Fellner M et al.; This contribution presents a novel method for the direct integration of a-priori knowledge in a neural network and its application for the online determination of a secondary metabolite during industrial yeast fermentation . Hereby, existing system knowledge is integrated in an artificial neural network (ANN) by means of 'functional nodes' . A generalized backpropagation algorithm is presented . For illustration, a set of ordinary differential equations describing the diacetyl formation and degradation during the cultivation is incorporated in a functional node and integrated in a dynamic feedforward neural network in a hybrid manner . The results show that a hybrid modelling approach exploiting available a-priori knowledge and experimental data can considerably outperform a pure data-based modelling approach with respect to robustness, generalization and necessary amount of training data . The number of training sets were decreased by 50%, obtaining the same accuracy as in a conventional approach . All incorrect decisions, according to defined cost criteria obtained with the conventional ANN, were avoided. Bioprocess Biosyst Eng, 2003 Jan, 25(4), 217 - 20 Epub 2002 Oct 16. Stabilizing feedback of a nonlinear process involving uncertain data; Krastanov MI et al.; A nonlinear control system describing the process of continuous methane fermentation is considered . Assuming that the parameters of the model are not exactly known but bounded within intervals, the set of optimal static points according to a practical criterion is computed . A continuous feedback control is proposed, which asymptotically stabilizes the dynamic system towards this set . Numerical results are also reported. J Chromatogr A, 2003 Aug 22, 1010(1), 63 - 74 Interlaboratory study of a liquid chromatography method for erythromycin: determination of uncertainty; Dehouck P et al.; Erythromycin is a mixture of macrolide antibiotics produced by Saccharopolyspora erythreas during fermentation . A new method for the analysis of erythromycin by liquid chromatography has previously been developed . It makes use of an Astec C18 polymeric column . After validation in one laboratory, the method was now validated in an interlaboratory study . Validation studies are commonly used to test the fitness of the analytical method prior to its use for routine quality testing . The data derived in the interlaboratory study can be used to make an uncertainty statement as well . The relationship between validation and uncertainty statement is not clear for many analysts and there is a need to show how the existing data, derived during validation, can be used in practice . Eight laboratories participated in this interlaboratory study . The set-up allowed the determination of the repeatability variance, s(2)r and the between-laboratory variance, s(2)L . Combination of s(2)r and s(2)L results in the reproducibility variance s(2)R . It has been shown how these data can be used in future by a single laboratory that wants to make an uncertainty statement concerning the same analysis. J Ind Microbiol Biotechnol, 2003 Oct, 30(10), 582 - 8 Epub 2003 Sep 09. A method for the selection of production media for actinomycete strains based on their metabolite HPLC profiles; R Tormo J et al.; The manipulation of growth conditions of microorganisms is a common strategy used by pharmaceutical companies to improve the quantities and spectra of secondary metabolites with potential therapeutic interest . In this work, the effects of fermentation media on secondary metabolite production from a set of Actinomycetes was statistically compared . For this purpose, we created an automated method for comparing the ability of microorganisms to produce different secondary metabolites . HPLC analyses guided the selection of those media in which a wider chemical diversity was obtained from microorganisms inoculated in a wide spectrum of production media . Fermented media yielding a better secondary metabolite profile were included in subsequent drug discovery screening. J Ind Microbiol Biotechnol, 2003 Oct, 30(10), 623 - 6 Epub 2003 Sep 11. Resistance to androstanes as an approach for androstandienedione yield enhancement in industrial mycobacteria; Perez C et al.; The resistance to androstandienedione (ADD) of industrial mycobacteria was demonstrated as a valuable approach to increasing ADD yield in sterol fermentations . Colonies growing at 1 mg/ml ADD in culture medium after nitrosoguanidine mutagenesis showed a differential behavior in respect to parentals in cholesterol biotransformation . In the presence of exogenous ADD, a substantial depletion of ADD production was observed in parental strains B3683 and Ex4, whereas it was unaffected, and even increased, in resistant colonies . An apparent reduction from ADD to androstandione and testosterone was also noticed . Furthermore, the ADD resistance phenotype may be related to the increase in steroid 1,2 dehydrogenase activity. Bioprocess Biosyst Eng, 2003 Jul, 25(6), 357 - 63 Epub 2003 Feb 19. Recovery of lactic acid from simultaneous saccharification and fermentation media using anion exchange resins; Moldes AB et al.; The physicochemical properties (capacity, kinetics and selectivity) of the ion exchange resins Amberlite IRA900, IRA400, IRA96 and IRA67 were determined to evaluate their comparative suitability for lactic acid recovery . Both the kinetics of lactic acid sorption from aqueous solutions and the equilibrium were assessed using mathematical models, which provided a close interpretation of the experimental results . The best resins (Amberlite IRA96 and IRA67) were employed in further fixed-bed operation using aqueous lactic acid solutions as feed . In this set of experiments, parameters such as capacity, regenerant consumption, percentage of lactic acid recovery and product concentration were measured . Amberlite IRA67, a weak base resin, was selected for lactic acid recovery from SSF (simultaneous saccharification and fermentation) broths . Owing to the presence of nutrients and ions other than lactate, a slightly decreased capacity was determined when using SSF media instead aqueous lactic acid solutions, but quantitative lactic acid recoveries at constant capacities were obtained in four sequential load/regeneration cycles. J Radiat Res (Tokyo), 2003 Jun, 44(2), 111 - 5 The protective effect of fermented milk kefir on radiation-induced apoptosis in colonic crypt cells of rats; Matsuu M et al.; To evaluate the effect of fermented milk kefir on X-ray-induced apoptosis in the colon of rats, we examined the apoptotic index, the mean number of apoptotic cells detected by H&E staining per crypt in the colon, in control rats and kefir-pretreated rats drinking kefir for 12 days before irradiation . Apoptotic cells were confirmed by TUNEL staining, and active caspase-3 expression was studied by immunohistochemistry . The cell position of apoptotic cells and active caspase-3 positive cells were examined . The apoptotic index of kefir-treated rats was significantly (p < 0.05) decreased 2 h after 1 Gy irradiation in comparison with control rats at crypt cell positions 1-3, 5-7, 13, and 15 . Active caspase-3 expression in the kefir-treated rats was also significantly (p < 0.05) reduced in comparison with control rats 2 h after 1 Gy irradiation at crypt cell positions 1-4, 13, and 15 . This study indicated that kefir protects colonic crypt cells against radiation-induced apoptosis, which was most pronounced in the stem cell region of the crypt . The antiapoptotic effect of fermented milk kefir was due to the inhibition of caspase-3 activation. Acta Pol Pharm, 2003 Mar-Apr, 60(2), 103 - 5 Quantification of p21 gene expression in Caco-2 cells treated with sodium butyrate using real-time reverse transcription-PCR (RT-PCR) assay; Orchel A et al.; Butyric acid, a short chain fatty-acid derived from bacterial fermentation of complex carbohydrates in the large intestine has been shown to be a growth inhibitory in many colon cancer cell lines . Butyrate induced inhibition of cellular proliferation is considered to result from the induction of P21 gene expression through the activation of this gene transcription . P21 is an inhibitor of cyclin-dependent protein kinases that are required for the cells to enter the DNA synthesis phase . In the present study the kinetics of the changes of the P21 transcription in Caco-2 colon adenocarcinoma cells treated with various concentrations of sodium butyrate was determined using a novel real-time quantitative RT-PCR (TaqMan) technique . Beta-actin mRNA and GAPDH mRNA levels were used as the endogenous references . Colonocytes were incubated with sodium butyrate at concentrations of 5 mM, 10 mM and 20 mM for 3, 6, 12, 24 and 48 h . The results of this study indicated that butyrate strongly induced P21 gene expression as early as 3 h after treatment . Characteristic patterns of time-dependent changes of the target gene expression were observed . The increases in P21 mRNA level were generally more pronounced at higher butyrate concentrations . Because Caco-2 cells are lacking the wild allele of the P53 gene, the present results support the hypothesis that butyrate induces P21 gene expression by P53-independent mechanism. J Chromatogr A, 2003 Aug 15, 1009(1-2), 189 - 96 Methodology for predicting the separation of proteins by hydrophobic interaction chromatography and its application to a cell extract; Lienqueo ME et al.; Hydrophobic interaction chromatography (HIC) is widely used in the downstream processing of proteins . Resolution of HIC is very good, but sometimes not as high as expected . Resolution values could be increased if good operating conditions were selected . In this paper we present a methodology for selecting good operating conditions . First, it is necessary to predict the dimensionless retention time (DRT) of each protein in the mixture . We use a correlation such as DRT = A + Bphi + Cphi2, where phi is the superficial hydrophobicity of the protein, which is calculated considering the hydrophobicity of the superficial amino acids using the Miyazawa-Jernigan scale . Considering that there was little interaction amongst proteins in a mixture at the concentrations investigated (2 g/l of each protein), the behaviour of the proteins in the mixture was considered to be similar to that of the individual proteins . Using simulations it was possible to test different operating conditions for the purification of a target protein from a mixture of proteins and it was possible to select ideal conditions . The methodology developed was also tested for the purification of a recombinant protein from a fermentation extract of yeast producing human superoxide dismutase and the results have been satisfactory. Int J Syst Evol Microbiol, 2003 Sep, 53(Pt 5), 1397 - 404 Sporomusa aerivorans sp . nov., an oxygen-reducing homoacetogenic bacterium from the gut of a soil-feeding termite; Boga HI et al.; Previously undescribed, homoacetogenic bacteria were isolated from gut homogenates of the soil-feeding termite Thoracotermes macrothorax . The isolates were slightly curved, banana-shaped rods (0.6-0.7x1.3-7.0 micro m) and were motile by one or more lateral flagella . In older cultures, cells formed club-like sporangia that developed into terminal, heat-resistant endospores . Cells stained Gram-positive but were Gram-negative in the KOH test . The isolates were mesophilic and grew homoacetogenically on H(2)/CO(2) and L-lactate . Strain TmAO3(T), which was characterized further, also grew homoacetogenically on pyruvate, citrate, L-alanine, D-mannitol, ethanol, formate and methanol . Succinate was decarboxylated to propionate; fumarate, L-malate and oxaloacetate were fermented to propionate and acetate . Hexoses were not used as substrates . Resting cells had a large capacity for hydrogen-dependent oxygen reduction {826 nmol min(-1) (mg protein)(-1)}, which enabled them to initiate growth in non-reduced basal medium that originally contained up to 1.5 kPa oxygen in the headspace, although growth commenced only after the medium had been rendered anoxic . Redox difference spectra of cell extracts indicated the presence of membrane-bound b-type cytochrome(s) . Comparative 16S rRNA gene sequence analysis revealed that strain TmAO3(T) belongs to a subgroup of the phylum of Gram-positive bacteria that is characterized by low DNA G+C content and a Gram-negative cell wall . It is related most closely to representatives of the genus SPOROMUSA: Based on morphological and physiological properties and on 16S rRNA gene sequence similarity of 94-97 % to other Sporomusa species, the isolates are assigned to Sporomusa aerivorans sp . nov . (type strain, TmAO3(T)=DSM 13326(T)=ATCC BAA-625(T)). Br J Nutr, 2003 Sep, 90(3), 625 - 34 Influence of diet on growth yields of rumen micro-organisms in vitro and in vivo: influence on growth yield of variable carbon fluxes to fermentation products; Blummel M et al.; The efficiency of rumen microbial production (EMP) in vitro and in vivo was examined for three roughages (lucerne (Medicago sativa L.) hay, oat (Avenia sativa L.)-berseem clover (Trifolium alexandrinum cultivar BigBee) hay and maize (Zea mays L.) crop residue (MCR)) and for five isonitrogenous (106 g crude protein (Nx6.25)/kg) diets formulated from lucerne hay, oat-berseem clover hay, MCR, soybean meal and maize grain to provide degradable intake protein for the production of 130 g microbial protein/kg total digestible nutrients . EMP in vivo was determined by intestinal purine recovery in sheep and ranged from 240 to 360 g microbial biomass/kg organic matter truly degraded in MCR and in one of the diets respectively (P<0.05) . EMP in vitro was estimated by the substrate degraded : gas volume produced thereby (termed partitioning factor, PF (mg/ml)) at times of estimated peak microbial production and after 16.0 and 24.0 h of incubation . For the diets, PF values were significantly related to EMP in vivo at peak microbial production (P=0.04), but not after 16.0 (P=0.08) and 24.0 h (P=0.66) . For roughages, PF values were significantly related to EMP in vivo only when measured after 16.0 h (P=0.04) . For MCR and diets, a close non-linear relationship was found between PF values at peak microbial production and EMP in vivo (R(2) 0.99, P<0.0001) suggesting a maximum EMP in vivo of 0.39 . Low gas production per unit substrate degraded (high PF) was associated with high EMP in vivo . The in vitro study of the products of fermentation, short-chain fatty acids, gases and microbial biomass (by purine analysis) after 16.0 h of incubation showed very strong relationships (R(2)> or =0.89, P<0.0001) between short-chain fatty acids, gases and gravimetrically measured apparent degradability . Except for maize grain, the true degradability of organic matter estimated by neutral-detergent solution treatment agreed with the sum of the products of fermentation (R(2) 0.81, P=0.0004) . After 16.0 h of incubation, the synergistic effects of diet ingredient on diets were greater for microbial biomass (18 %) than for short-chain fatty acids and gas production (7 %) . It is concluded that measurement of gas production only gives incomplete information about fodder quality; complementation of gas measurements by true degradability measurements is recommended. Br J Nutr, 2003 Sep, 90(3), 617 - 23 Influence of different concentrations of disodium fumarate on methane production and fermentation of concentrate feeds by rumen micro-organisms in vitro; Carro MD et al.; Batch cultures of mixed rumen micro-organisms were used to study the effects of different concentrations of disodium fumarate on the fermentation of five concentrate feeds (maize, barley, wheat, sorghum and cassava meal) . Rumen contents were collected from four Merino sheep fed lucerne hay ad libitum and supplemented with 300 g concentrate/d . Disodium fumarate was added to the incubation bottles to achieve final concentrations of 0, 4, 7 and 10 mM-fumarate . In 17 h incubations, the final pH and total volatile fatty acid production increased (P<0.001) linearly for all substrates as fumarate concentration increased from 0 to 10 mm . Propionate and acetate production increased (P<0.05), while the value of the acetate:propionate ratio decreased (P<0.05) linearly with increasing doses of fumarate . In contrast, l-lactate and NH3-N concentrations in the cultures were not affected (P>0.05) by the addition of fumarate . For all substrates, fumarate treatment decreased (P<0.05) CH4 production, the mean values of the decrease being 2.3, 3.8 and 4.8 % for concentrations of 4, 7 and 10 mM-fumarate respectively . Addition of fumarate did not affect (P>0.05) the total gas production . If the results of the present experiment are confirmed in vivo, fumarate could be used as a feed additive for ruminant animals fed high proportions of cereal grains, because it increased pH, acetate and propionate production and it decreased CH4 production. J Agric Food Chem, 2003 Sep 24, 51(20), 5877 - 81 Direct method for determining seed and skin proanthocyanidin extraction into red wine; Peyrot des Gachons C et al.; A method was developed for determining the amount of seed and skin proanthocyanidin extraction into wines by direct measurement . This method was based upon the analysis of proanthocyanidin cleavage products after acid catalysis in the presence of excess phloroglucinol . On the basis of the analysis of proanthocyanidin extracts from grape tissues, two observations were made as follows: (i) the seed and skin proanthocyanidin extension subunit compositions were considerably different from each other, and (ii) their composition did not vary with extraction time . Thus, by comparing the proportional extension subunit composition of proanthocyanidins in wine relative to their proportional composition in corresponding grape seed and skin, it was possible to determine the contribution of each to wine . To provide additional information, the procedure was used to investigate seed and skin proanthocyanidin extraction during commercial-scale fermentations that had undergone 4 or 10 day low temperature prefermentation skin contact prior to the onset of fermentation . The results for both fermentations indicated that the proportion of skin tannin declined during fermentation and also showed that at the end of fermentation the amount and proportion of skin tannin were the same. Ying Yong Sheng Tai Xue Bao, 2003 Jun, 14(6), 1029 - 30 {Application of Vc fermentation waste residue on prevention and control of plant diseases in protective ground}; Zhu K et al.; The prevention and control of tomato plant diseases were conducted in protective ground using Vc fermentation waste residue treated by enzymolysis and ultrasonic wave . The results showed that the seedlings planted for 3 weeks on the protective ground soil continuously cropped tomato plant for 9 years and fertilized 75, 150 and 300 kg.hm-2 grew well . Their biomass were increased by 123%, 164% and 182%, and the disease incidence rates were decreased by 59%, 78% and 85%, respectively . Under application of 300 kg.hm-2 Vc fermentation waste residue, the products of tomato grown for 10 weeks on the soil continuously cropped tomato plant for 9, 6 and 2 years were increased by 60%, 43% and 14%, respectively, and the disease incidence rates were all decreased by 50%. Zhejiang Da Xue Xue Bao Yi Xue Ban, 2003 Aug, 32(4), 292 - 5 {Inhibition of Cryptoporus volvatus ferment substance on release of leukotriene B4, C4 and D4 from neutrophils in rats in vitro}; Jin SH et al.; OBJECTIVE: To study inhibitory the effects of Cryptoporus volvatus ferment substance(CVFS) on leukotriene production in vitro from neutrophils in rats . METHODS: Neutrophil aggregation was induced by intraperitoneal injection of glycogen in rats . After 16 h, intraperitoneal lavage fluid(PLF) was collected and neutrophils were removed . Neutrophils were stimulated by calcium ionophore A23187 in vitro to produce leukotriene B(4), C(4), D(4) . The concentrations of leukotriene B(4), C(4) and D(4) were measured by reversed-phase high-performance liquid chromatography(HPLC) . RESULT: CVFS at 0.25, 1, 4 mg x L(-1)decreased leukotriene B(4), C(4), D(4) release from neutrophils in a concentration-dependent manner . Inhibitory rate of CVFS 0.25, 1, 4 mg x L(-1 )on A23187-induced leukotriene B(4) production was 27.4%, 54.2% and 78.8%(P<0.05), respectively . Inhibitory rate of leukotriene C(4) production was 65.1%, 74.3 and 79.0%(P<0.05), respectively . Inhibitory rate of leukotriene D(4) production was 55.6%, 60.9% and 72.8%(P<0.05), respectively . CONCLUSION: The results suggest that suppression of leukotriene release may be a mechanism of the anti-inflammation and anti-asthma effects of CVFS. Gut, 2003 Oct, 52(10), 1442 - 7 Short chain fatty acids stimulate epithelial mucin 2 expression through differential effects on prostaglandin E(1) and E(2) production by intestinal myofibroblasts; Willemsen LE et al.; BACKGROUND: The mucus layer protects the gastrointestinal mucosa from mechanical, chemical, and microbial challenge . Mucin 2 (MUC-2) is the most prominent mucin secreted by intestinal epithelial cells . There is accumulating evidence that subepithelial myofibroblasts regulate intestinal epithelial cell function and are an important source of prostaglandins (PG) . PG enhance mucin secretion and are key players in mucoprotection . The role of bacterial fermentation products in these processes deserves further attention . AIMS: We therefore determined whether the effect of short chain fatty acids (SCFA) on MUC-2 expression involves intermediate PG production . METHODS: Both mono- and cocultures of epithelial cells and myofibroblasts were used to study the effects of SCFA on MUC-2 expression and PG synthesis . Cell culture supernatants were used to determine the role of myofibroblast derived prostaglandins in increasing MUC-2 expression in epithelial cells . RESULTS: Prostaglandin E(1) (PGE(1)) was found to be far more potent than PGE(2) in stimulating MUC-2 expression . SCFA supported a mucoprotective PG profile, reflected by an increased PGE(1)/PGE(2) ratio in myofibroblast supernatants and increased MUC-2 expression in mono- and cocultures . Incubation with indomethacin revealed the latter to be mediated by PG . CONCLUSIONS: SCFA can differentially regulate PG production, thus stimulating MUC-2 expression in intestinal epithelial cells . This mechanism involving functional interaction between myofibroblasts and epithelial cells may play an important role in the mucoprotective effect of bacterial fermentation products. J Appl Microbiol, 2003, 95(4), 847 - 52 Effects of xylulokinase activity on ethanol production from D-xylulose by recombinant Saccharomyces cerevisiae; Lee TH et al.; AIMS: Recombinant Saccharomyces cerevisiae strains harbouring different levels of xylulokinase (XK) activity and effects of XK activity on utilization of xylulose were studied in batch and fed-batch cultures . METHODS AND RESULTS: The cloned xylulokinase gene (XKS1) from S . cerevisiae was expressed under the control of the glyceraldehyde 3-phosphate dehydrogenase promoter and terminator . Specific xylulose consumption rate was enhanced by the increased specific XK activity, resulting from the introduction of the XKS1 into S . cerevisiae . In batch and fed-batch cultivations, the recombinant strains resulted in twofold higher ethanol concentration and 5.3- to six-fold improvement in the ethanol production rate compared with the host strain S . cerevisiae . CONCLUSIONS: An effective conversion of xylulose to xylulose 5-phosphate catalysed by XK in S . cerevisiae was considered to be essential for the development of an efficient and accelerated ethanol fermentation process from xylulose . SIGNIFICANCE AND IMPACT OF THE STUDY: Overexpression of the XKS1 gene made xylulose fermentation process accelerated to produce ethanol through the pentose phosphate pathway. J Anim Sci, 2003 Sep, 81(9), 2247 - 58 Performance of sows fed high levels of nonstarch polysaccharides during gestation and lactation over three parities; van der Peet-Schwering CM et al.; The effect of feeding sows a starch diet or a diet with a high level of nonstarch polysaccharides (NSP) during gestation, lactation, or both gestation and lactation during the first three parities on reproductive performance, body weight, and backfat was studied . Four-hundred and forty-four postpuberal gilts were allotted to a 2 x 2 x 2 factorial experiment . Treatments were diet composition during gestation (including the weaning-to-estrus interval; G-Starch: 274 g/kg of starch and 123 g/kg of fermentable NSP or G-NSP: 86 g/kg of starch and 300 g/kg of fermentable NSP), diet composition during lactation (L-Starch: 293 g/kg of starch and 113 g/kg of fermentable NSP or L-NSP: 189 g/kg of starch and 216 g/kg of fermentable NSP) and group-housing system during gestation (free access stalls or electronic feeding) . Both gestation diets were formulated to be isoenergetic . During lactation, sows were given free access to the lactation diets from d 6 after parturition onwards . Body weight and backfat gains during gestation were lower in sows fed the G-NSP diet than in those fed the G-starch diet (P < 0.001) . The effects were more pronounced in the electronic feeding system than in the free access stalls . These results indicate an overestimation of the energy value of fermentable NSP . Body weight and backfat losses during lactation were less in sows fed the G-NSP diet during gestation than in those fed the G-starch diet (P < 0.05),which can be explained by a 0.4 kg/d higher (P < 0.001) feed intake during lactation of the sows fed the G-NSP diet . Sows fed the L-NSP diet lost more backfat during lactation than sows fed the L-starch diet (P < 0.05) . The number of total piglets born and live-born piglets was 0.5 piglet higher in sows fed the G-NSP diet than in those fed the G-starch diet (P < 0.05) . Lactation diet did not affect the number of total piglets born or live-born piglets . This study shows that, although high NSP diets negatively influence body weight and backfat thickness of the sows, it is possible to feed sows a diet with a high level of fermentable NSP diet during both gestation and lactation without negative effects on reproductive performance . Under the conditions of this study, feeding sows a diet with a high level of fermentable NSP during gestation and a high level of starch during lactation seems the most favorable feeding strategy. J Anim Sci, 2003 Sep, 81(9), 2131 - 8 Swine manure composition affects the biochemical origins, composition, and accumulation of odorous compounds; Miller DN et al.; Odors from swine production facilities are associated with the storage and decomposition of manure . Diet is linked to manure composition and will likely affect odor, but the microbial mechanisms responsible for manure decomposition and odor production are poorly understood . To identify the sources of odor during manure fermentation, substrates (starch, casein, and cellulose) were added to slurries of fresh swine manure, and the anaerobic accumulation of fermentation products and the consumption of substrates were measured relative to no addition of substrates . Volatile fatty acids and alcohols were the dominant fermentation products in all treatments . The total VFA concentration from starch treatment was greater (P < 0.001) than for all other treatments . Branched-chain VFA and aromatic compounds accumulated in all treatments, but accumulation in the casein treatments was greater (P < 0.001) than in all other treatments . Thus, addition of carbohydrate to swine manure slurries did not circumvent protein fermentation, as was previously observed in cattle manure slurries . Based on substrate loss, starch and protein fermentation were equivalent in all treatments, with losses of each exceeding 4% of the DM . Substrate additions had a limited effect on the overall accumulation of odor compounds in manure and on odor compound composition . Compared with the results of the earlier fermentation study of fresh cattle manure, swine manure fermentation produced less lactate and more products of protein fermentation (branched-chain VFA and aromatic ring compounds) . We hypothesize that differences in manure organic matter composition between cattle and swine, a result of diet and digestion, select for bacterial communities that are adapted to the available substrate composition. Org Biomol Chem, 2003 Aug 21, 1(16), 2840 - 7 Direct production of ivermectin-like drugs after domain exchange in the avermectin polyketide synthase of Streptomyces avermitilis ATCC31272; Gaisser S et al.; Ivermectin, a mixture of 22,23-dihydroavermectin B1a9 with minor amounts of 22,23-dihydroavermectin B1b 10, is one of the most successful veterinary antiparasitic drugs ever produced . In humans, ivermectin has been used for the treatment of African river blindness (onchocerciasis) resulting in an encouraging decrease in the prevalence of skin and eye diseases linked to this infection . The components of ivermectin are currently synthesized by chemical hydrogenation of a specific double bond at C22-C23 in the polyketide macrolides avermectins B1a 5 and B1b 6, broad-spectrum antiparasitic agents isolated from the soil bacterium Streptomyces avermitilis . We describe here the production of such compounds (22,23-dihydroavermectins B1a 9 and A1a 11) by direct