Competition for glucose between the yeasts Saccharomyces cerevisiae and Candida utilis.

The competition between the yeasts Saccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621 for glucose was studied in sugar-limited chemostat cultures. Under aerobic conditions, C. utilis always successfully completed against S. cerevisiae. Only under anaerobic conditions did S. cerevisiae become the dominant species. The rationale behind these observations probably is that under aerobic glucose-limited conditions, high-affinity glucose/proton symporters are present in C. utilis, whereas in S. cerevisiae, glucose transport occurs via facilitated diffusion with low-affinity carriers. Our results explain the frequent occurrence of infections by Crabtree-negative yeasts during bakers' yeast production.     

Dot assay for determining adhesive interactions between yeasts and bacteria under controlled hydrodynamic conditions

Candida belongs to the normal human microflora and are found adhering to a number of human body tissues as well as to a variety of biomaterials implants. Often, yeasts adhere in association with bacteria, but to date there is no definitive assay to investigate adhesive interactions between yeasts and bacteria adhering on surfaces. Although we recently described the use of a parallel plate flow chamber to this purpose [Millsap, K.W., Bos, R., Van der Mei, H.C., Busscher, H.J., 1998. Adhesive interactions between medically important yeasts and bacteria. FEMS Microbiol. Rev. 21, 321–336], the method was slow and evaluation of a large number of strains showed major biological variation between experiments. Here, we describe a new assay for the simultaneous determination of the adhesive interactions between yeasts and different bacterial strains on a surface under controlled hydrodynamic conditions. On an acrylic surface, the presence of adhering bacteria suppressed adhesion of Candida albicans ATCC 10261 to various degrees, depending on the bacterial strain involved. Suppression of C. albicans ATCC 10261 adhesion was strongest by Actinomyces naeslundii T14V-J1, while adhering Streptococcus gordonii NCTC 7869 caused the weakest suppression of yeast adhesion. When adhering yeasts and bacteria were challenged with the high detachment force of a passing liquid–air interface, the majority of the yeasts detached, while C. albicans adhering on the control, bare polymethylmethacrylate surface formed aggregates. Summarizing, this study presents a new method to determine suggested adhesive interactions between yeasts and adhering bacteria under controlled hydrodynamic conditions. However, the results seem to indicate that these adhesive interactions may well not exist, but that instead different bacterial strains have varying abilities to discourage yeast adhesion.     

Aggregation and separability of the shikimate pathway enzymes in yeasts

Gel filtration was employed to estimate the molecular weights and to determine possible physical aggregation of enzymes [5-dehydroquinate synthase (DHQ synthase), 5-dehydroquinase (DHQase, EC 4.2.1.10), shikimate: NADP oxidoreductase (EC 1.1.1.25), shikimate kinase (EC 2.7.1.71), 3-enolpyruvylshikimate 5-phosphate synthase (EPSP synthase)] in the shikimate pathway in eleven species of yeasts. The five enzymes were not aggregated in extracts of Hansenula henricii, H. fabianii, H. anomala, Candida utilis, Pichia guilliermondii, and Lodderomyces elongisporus. Two enzymes (DHQase and shikimate:NADP oxidoreductase) were not separable by this method and by ion exchange chromatography, and we conclude that they exist as an aggregate in these yeasts. Evidence is presented for an enzyme aggregate containing five activities, with a molecular weight of approximately 280,000 in Rhodosporidium spaerocarpum, Rh. toruloides, Rhodotorula rubra, Saccharomycopsis lipolytica, and Saccharomyces cerevisiae. Similarities between the enzymes in the shikimate pathway of plants, bacteria, and other fungi and those of investigated yeasts are discussed.     

Carnitine acetyltransferase activity in oleaginous yeasts

Abstract The highest activities of carnitine acetyltransferase (CAT) were found in non-oleaginous yeasts ( Candida utilis and Saccharomyces cerevisiae ); lower activities, ranging from 50% down to 3% of the highest values, were found in various strains of oleaginous yeasts ( Candida curvata, Lipomyces starkeyi, Rhodosporidium toruloides and Trichosporon cutaneum ). Supply of acetyl units into the cytosol of the latter, but not of the former yeasts, was therefore necessarily reliant on the action of ATP: citrate lyase (ACL), which was present in all oleaginous yeasts. There was no correlation, however, between the amount of lipid in the oleaginous yeasts and the specific activities of either CAT or ACL. Activity of CAT was increased up to 30-fold by growing yeasts on a triacylglycerol.     

Yeasts in molecular biology. Spheroplast preparation withCanadida utilis,Schizosaccharomyces pombe andSaccharomyces cerevisiae

Efficient preparation of spheroplasts fromCandida utilis, Saccharomyces cerevisiae, andSchizosaccharomyces pombe, using a purified mixture of enzymes fromTrichoderma harzianum, is described. Limitations of other methods, and differences between yeasts are demonstrated.     

A biochemical explanation for lipid accumulation in Candida 107 and other oleaginous micro-organisms.

The biochemical explanation for lipid accumulation was investigated principally in Candida 107 and, for comparison, in the non-oleaginous yeast Candida utilis. There were no significant differences between these two yeasts in their control of glucose uptake; in both yeasts, the rates of glucose uptake were independent of the growth rate and were higher in carbon-limited chemostat cultures than in nitrogen-limited cultures. There was no lipid turnover in either yeast, as judged from [14C]acetate uptake and subsequent loss of 14C from the lipid of steady-state chemostat cultures. Acetyl-CoA carboxylase from both yeasts was similar in most characteristics except that from Candida 107 was activated by citrate (40% activation at 1 mM). The enzyme from Candida 107 was relatively unstable and, when isolated from nitrogen-limited (lipid-accumulating) cultures, was accompanied by a low molecular weight inhibitor. The reason for lipid accumulation is attributed to the decrease in the intracellular concentration of AMP as cultures become depleted of nitrogen. As the NAD+-dependent isocitrate dehydrogenase of Candida 107, but not C. utilis, requires AMP for activity, the metabolism of citrate through the tricarboxylic acid cycle in the mitochondria becomes arrested. In Candida 107, but not in C. utilis, there is an active ATP:citrate lyase which converts the accumulating citrate, when it passes into the cytosol, into acetyl-CoA and oxaloacetate. The former product is then available for fatty acid biosynthesis which is stimulated by the high ATP concentration within the cells, by the activation of acetyl-CoA carboxylase by citrate and by the provision of NADPH generated as oxaloacetate is converted via malate to pyruvate. Similar characteristics were evident in oleaginous strains of Rhodotorula glutinis and Mucor circinelloides but not in non-oleaginous representatives of these species.     

Energy supply and cell yield in aerobically growth microorganisms

Cell yields of Pseudomonas fluorescens, Aerobacter cloacae UW-C83, Escherichia coli K-12, Candida utilis UW-3, and Saccharomyces carlsbergensis UW-110 grown aerobically on several carbon sources are reported. The Y(atp) concept is discussed in relation to the yields on oxygen and to assumed P to O ratios for bacteria and reported P to O ratios for yeast.     

Lytic activity against yeasts in Bacillus brevis culture broth

The cultural broth of Bacillus brevis, strain 5/4, is capable of lysing the cell walls of Rhodotorula rubra, Candida utilis and other yeasts. The lytic action of the cultural broth is ascribed to proteolysis.     

Adhesion and surface-aggregation of Candida albicans from saliva on acrylic surfaces with adhering bacteria as studied in a parallel plate flow chamber

Adhesive interactions between Candida albicans and oral bacteria are generally thought to play a crucial role in the microbial colonization of denture acrylic, which may lead to denture stomatitis. This study investigated the influence of saliva on the adhesive interactions between C. albicans and Streptococcus sanguis or Actinomyces naeslundii on denture acrylic. First, bacteria were allowed to adhere to the acrylic surface from a flowing suspension, and subsequently yeasts were flowed over the acrylic surface. The organisms were assayed in the presence or absence of human whole saliva. All experiments were carried out in a parallel plate flow chamber and enumeration was done in situ with an image analysis system. In the absence of adhering bacteria, adhesion of C. albicans from buffer was more extensive than from saliva. However, in the presence of adhering bacteria, yeast adhesion from saliva was increased with respect to adhesion of yeasts from buffer, indicating that specific salivary components constitute a bridge between bacteria and yeasts. In all cases, yeast aggregates consisting of 3 to 5 yeast cells were observed adhering to the surface. A surface physico-chemical analysis of the microbial cell surfaces prior to and after bathing the microorganisms in saliva, suggests that this bridging is mediated by acid-base interactions since all strains show a major increase in electron-donating surface free energy parameters upon bathing in saliva, with no change in their zeta potentials. The surface physico-chemical analysis furthermore suggests that S. sanguis and A. naeslundii may use a different mechanism for adhesive interactions with C. albicans in saliva.     

Biomass production of yeast isolate from salad oil manufacturing wastewater

Conversion of oil-rich salad oil manufacturing wastewater (SOMW) into protein source for animal feed through biomass production of yeast isolate was investigated in this study. Five species of yeasts, including Rhodotorula rubra, Candida tropicalis, C. utilis, C. boidinii, Trichosporon cutaneum, were isolated from SOMW following enrichment culture. Of them, C. utilis was chosen as the sole biomass producer in the study due to its greatest oil uptake rate, 0.96 kg oil kg(-1) biomass d(-1), and highest specific growth rate, 0.25 h(-1). The cells of C. utilis contained 26% protein, 9% crude lipid, 55% carbohydrate and balanced amino acid compositions. The initial N:C ratio in SOMW drastically influenced oil reduction efficiency, biomass production and protein content of C. utilis, and therefore a range between 1:6 and 1:8 was recommended in consideration of these three factors simultaneously.     

Temperature adaptation in yeasts: the role of fatty acids

Studies on the yeasts Candida oleophila, Candida utilis, Lipomyces starkeyi, Rhodosporidium toruloides and Saccharomyces cerevisiae revealed the existence of three different temperature adaptation responses involving changes in fatty acid composition. These conclusions were drawn by determining the growth rates, total cellular fatty acid content, fatty acid composition, degree of unsaturation, and the mean chain length of fatty acids over a range of growth temperatures. Within temperatures permitting growth, there were no changes in the major fatty acids of any of the yeasts, but the absolute amounts and relative compositions of the fatty acids did alter. In S. cerevisiae there were temperature-induced changes in the mean fatty acid chain length, whereas in R. toruloides there were changes in the degree of unsaturation. C. oleophila, C. utilis and L. starkeyi showed both responses, depending on whether the growth temperature was above or below 20-26 degrees C. Below 20-26 degrees C temperature-dependent changes were observed in the mean chain length whereas above 20-26 degrees C there were changes in the degree of unsaturation.     

Yeast pyruvate decarboxylases: variation in biocatalytic characteristics for (R)-phenylacetylcarbinol production

Based on previous studies, Candida utilis pyruvate decarboxylase (PDC) proved to be a stable and high productivity enzyme for the production (R)-phenylacetylcarbinol (PAC), a pharmaceutical precursor. However, a portion of the substrate pyruvate was lost to by-product formation. To identify a source of PDC which might overcome this problem, strains of four yeasts -- C. utilis, Candida tropicalis, Saccharomyces cerevisiae and Kluyveromyces marxianus -- were investigated for their PDC biocatalytic properties. Biotransformations were conducted with benzaldehyde and pyruvate as substrates and three experimental systems were employed (in the order of increasing benzaldehyde concentrations): (I) aqueous (soluble benzaldehyde), (II) aqueous/benzaldehyde emulsion, and (III) aqueous/octanol-benzaldehyde emulsion. Although C. utilis PDC resulted in the highest concentrations of PAC and was the most stable enzyme, C. tropicalis PDC was associated with the lowest acetoin formation. For example, in system (III) the ratio of PAC over acetoin was 35 g g(-1) for C. tropicalis PDC and 9.2 g g(-1) for C. utilis PDC. The study thereby opens up the potential to design a PDC with both high productivity and high yield characteristics.     

Microbial interactions during sugar cane must fermentation for bioethanol production: does quorum sensing play a role?

Microbial interactions represent important modulatory role in the dynamics of biological processes. During bioethanol production from sugar cane must, the presence of lactic acid bacteria (LAB) and wild yeasts is inevitable as they originate from the raw material and industrial environment. Increasing the concentration of ethanol, organic acids, and other extracellular metabolites in the fermentation must are revealed as wise strategies for survival by certain microorganisms. Despite this, the co-existence of LAB and yeasts in the fermentation vat and production of compounds such as organic acids and other extracellular metabolites result in reduction in the final yield of the bioethanol production process. In addition to the competition for nutrients, reduction of cellular viability of yeast strain responsible for fermentation, flocculation, biofilm formation, and changes in cell morphology are listed as important factors for reductions in productivity. Although these consequences are scientifically well established, there is still a gap about the physiological and molecular mechanisms governing these interactions. This review aims to discuss the potential occurrence of quorum sensing mechanisms between bacteria (mainly LAB) and yeasts and to highlight how the understanding of such mechanisms can result in very relevant and useful tools to benefit the biofuels industry and other sectors of biotechnology in which bacteria and yeast may co-exist in fermentation processes.     

Effects of suspension density on microbial metabolic processes

Respiration of yeasts Saccharomyces cerevisiae, Rhodotorula glutinis, Endomyces magnusii, and Candida utilis, and of bacteria Escherichia coli and Bacillus megaterium, anaerobic production of CO2 by S. cerevisiae, active transport of quinovose by R. glutinis and of L-proline and L-leucine by S. cerevisiae were highly dependent on cell suspension density. Respiration of S. cerevisiae in the presence of glucose decreased in a biphasic fashion from 140 to 40 nmol O2 per mg dry solid per min as suspension density increased from 0.01 to 2 mg/mL. Higher partial pressures of oxygen further enhanced the trend. The active transports were affected monophasically in the maximum rate of uptake which was as much as ten times greater at low than at high suspension densities. A component of the external medium is suspected to cause the decrease of metabolic functions at higher cell densities, acting as a noncompetitive inhibitor. The component was present and mutually active in suspensions of the various yeasts as well as of bacteria. Its properties and results of model experiments suggest it to be dissolved carbon dioxide.     

Evaluation of industrial yeasts for pathogenicity.

Eleven yeasts representative of species of industrial interest were compared with Candida albicans for their potential pathogenicity for untreated and cortisone-treated mice. Only C. tropicalis produced a progressive infection similar to that produced by C. albicans. Candida lipolytica, Torulopsis spp., and Hansenula polymorpha were not recovered from mice 6 days after inoculation. Kluyveromyces fragilis, C. pseudotropicalis, C. utilis, C. guilliermondii and C. maltosa were recovered from mice but did not produce evidence of infection.     

白假丝酵母与细菌相互作用机制及其与感染的关系

人体内定植着数目庞大、结构复杂的细菌及真菌等微生物群,它们之间存在复杂的相互作用。既往研究主要集中于细菌或真菌的某个单一物种,但最近研究表明,细菌与真菌之间的相互作用对更好地理解体内的微生态系统至关重要。白假丝酵母(又称白念珠菌)是人备体中最常见的机会性致病真菌,通常被认为是人类正常菌群的一部分。白念珠菌与细菌的相互作用近年来备受关注,其协同和拮抗作用有助于维持不同物种间复杂的平衡关系。了解白念珠菌与细菌的相互作用,不仅可加深对微生物致病机制的理解,还可为预防和治疗白念珠菌或细菌感染及新型抗菌药物研发提供新途径。本文就白念珠菌与细菌共存时的相互作用机制及其对人类健康、疾病的影响进行综述,从而为控制念珠菌或细菌感染提供新的策略。     

Novel bioemulsifiers from microorganisms for use in foods

The main objective of this study was to test a range of microorganisms for production of extracellular, high molecular weight emulsifiers for potential use in foods. A standard emulsification assay developed specifically for assessing food emulsifiers was used to examine 24 extracellular microbial products from bacteria, yeasts and algae. Of the 24 products tested, nine had emulsification ability that was as good as and eight had emulsifying properties that were better than those of the commonly used food emulsifiers gum arabic and carboxymethylcellulose. The eight good producer organisms included the yeasts Candida utilis, Candida valida, Hansenula anomala, Rhodospiridium diobovatum and Rhodotorula graminis, the red alga Porphiridium cruentum, and the bacteria Klebsiella spp. and Acinetobacter calcoaceticus. Of these, C. utilis was selected for further study due to the excellent emulsification properties of its extracellular products and the food-grade status of the organism. Crude preparations of the bioemulsifier from C. utilis exhibited low viscosity and had a carbohydrate content of over 80%. Preliminary trials showed that the bioemulsifier from this organism had potential for use in salad cream.     

Fructose-6-phosphate is not a substrate for glucose-6-phosphate dehydrogenase

D-Fructose-6-phosphate was shown not to be a substrate for glucose-6-phosphate dehydrogenases (EC. 1.1.1.49) from human erythrocytes, bovine adrenal, rat liver, three yeasts (brewer's yeast, baker's yeast, and Candida utilis), and Leuconostoc mesenteroides. These findings contrast with those of G.M. Kidder (J. Exp. Zool., 226:385-390, '83).     

Physical properties of microbial suspensions

Physical properties of suspensions of Saccharomyces cerevisiae, Candida utilis and Escherichia coli (density, viscosity and surface tension) were measured in synthetic suspensions formed of centrifuged biomass and supernatants from various stages of batch cultivation in the range from 0 to 10% w/v for yeasts and from 0 to 0.25% w/v for bacteria. Surface tension was also measured in native suspensions in the range of 0 less than or equal to Cm less than or equal to 2.0% w/v. All single cell suspensions were found to be Newtonian in behaviour. Densities strictly obey the mixing law, viscosities of Saccharomyces cerevisiae suspensions were correlated by an empirical relation in dependence of Cm and t, surface tensions were correlated graphically for suspensions of Saccharomyces cerevisiae and Escherichia coli, since experiments with both microorganisms have shown that the previously published approximate correlation can safely be used.     

Localization and kinetics of pyruvate-metabolizing enzymes in relation to aerobic alcoholic fermentation in Saccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621

The role of pyruvate metabolism in the triggering of aerobic, alcoholic fermentation in Saccharomyces cerevisiae has been studied. Since Candida utilis does not exhibit a Crabtree effect. this yeast was used as a reference organism. The localization, activity and kinetic properties of pyruvate carboxylase (EC 6.4.1.1), the pyruvate dehydrogenase complex and pyruvate decarboxylase (EC 4.1.1.1) in cells of glucose-limited chemostat cultures of the two yeasts were compared. In contrast to the general situation in fungi, plants and animals, pyruvate carboxylase was found to be a cytosolic enzyme in both yeasts. This implies that for anabolic processes, transport of C4-dicarboxylic acids into the mitochondria is required. Isolated mitochondria from both yeasts exhibited the same kinetics with respect to oxidation of malate. Also, the affinity of isolated mitochondria for pyruvate oxidation and the in situ activity of the pyruvate dehydrogenase complex was similar in both types of mitochondria. The activity of the cytosolic enzyme pyruvate decarboxylase in S. cerevisiae from glucose-limited chemostat cultures was 8-fold that in C. utilis. The enzyme was purified from both organisms, and its kinetic properties were determined. Pyruvate decarboxylase of both yeasts was competitively inhibited by inorganic phosphate. The enzyme of S. cerevisiae was more sensitive to this inhibitor than the enzyme of C. utilis. The in vivo role of phosphate inhibition of pyruvate decarboxylase upon transition of cells from glucose limitation to glucose excess and the associated triggering of alcoholic fermentation was investigated with 31P-NMR. In both yeasts this transition resulted in a rapid drop of the cytosolic inorganic phosphate concentration. It is concluded that the relief from phosphate inhibition does stimulate alcoholic fermentation, but it is not a prerequisite for pyruvate decarboxylase to become active in vivo. Rather, a high glycolytic flux and a high level of this enzyme are decisive for the occurrence of alcoholic fermentation after transfer of cells from glucose limitation to glucose excess.