Extracytoplasmic phosphatases of selected species of Saccharomyces, Kluyveromyces and Rhodotorula

Phosphatase activities of yeasts belonging to the genera Saccharomyces, Kluyveromyces and Rhodotorula were studied. Rhodotorula rubra exhibited activities at acid, neutral and alkaline pH; the other yeasts only had activity at acid pH. Growing yeasts in a constant pH (4.5) medium decreased phosphatase activities in Saccharomyces and Kluyveromyces, while neutral activity was enhanced in Rhodotorula rubra which excreted more enzyme under these conditions. Washing cells with sucrose solutions lowered phosphatase activities in all yeasts, due to enzyme liberation. Acid phosphatase activities in isolated and purified cell walls were very small. Phosphatases thus appear not to be strongly bound to yeast cell walls.     

Identification of yeasts of clinical interest on CHROMagar Candida culture medium.

The efficiency of CHROMagar Candida was evaluated as a medium for the presumptive identification of yeasts. We tested 36 different yeast species, pertaining to 9 genera: one Blastoschizomyces, 20 Candida, five Cryptococcus, two Geotrichum, one Kloeckera, two Pichia, three Rhodotorula, one Saccharomyces and one Trichosporon, to determine the colony colors and characteristics on this medium. Afterwards, we identified 2,230 strains isolated directly on CHROMagar Candida from clinical samples by specific colouration and morphology of the colonies after 72 hours. Their results were compared with standard methods for the identification of yeasts. The sensitivity and specificity were both superior to 97% for all strains, 100% and 100% for Candida albicans, 97.3% and 99.9% for Candida glabrata, 92.3% and 99.6% for Candida krusei, 90.3% and 99.6% for Candida parapsilosis, and 100% and 100% for Candida tropicalis. CHROMagar Candida is a very useful medium for the culture of clinical samples; its use for identification of yeasts has an accuracy of 97.5%, close to 100% of conventional methods.     

Glycolic acid production using ethylene glycol-oxidizing microorganisms.

Screening for microorganisms oxidizing ethylene glycol to glycolic acid was carried out. Among stock cultures, several yeasts and acetic acid bacteria showed high glycolic acid producing activity. Pichia naganishii AKU 4267 formed the highest concentration of glycolic acid, 35.3 g/l, from 10% (v/v) ethylene glycol (molar conversion yield, 26.0%). Among soil isolates, Rhodotorula sp. 3Pr-126, isolated using propylene glycol as a sole carbon source, formed the highest concentration of glycolic acid, 25.1 g/l, from 10% (v/v) ethylene glycol (molar conversion yield, 18.5%). Rhodotorula sp. 3Pr-126 showed higher activity toward 20% (v/v) ethylene glycol than P. naganishii AKU 4267. Optimization of the conditions for glycolic acid production was investigated using P. naganishii AKU 4267 and Rhodotorula sp. 3Pr-126. Under the optimized conditions, P. naganishii AKU 4267 and Rhodotorula sp. 3Pr-126 formed 105 and 110 g/l of glycolic acid (corrected molar conversion yields, 88.0 and 92.2%) during 120 h of reaction, respectively.     

Inhibition of Histoplasma capsulatum by Candida albicans and Other Yeasts on Sabouraud''s Agar Media

The inhibition of growth of Histoplasma capsulatum by Candida albicans and other yeasts on Sabouraud's agar was investigated. Histoplasma (yeast-phase inoculum) was grown alone and in mixtures with yeasts at 25 C for 4-week periods. As few as 10 colonies of C. albicans completely inhibited the growth of approximately 50,000 potential colonies of Histoplasma. The pH was determined in cultures of 36 colonies of Candida on media containing 1, 2, and 4% glucose by spotting the agar with pH indicators. A drop in the pH became noticeable in all three media about the 3rd day of incubation, and a pH of 3.5 was reached in about 7 days. Subsequently, the pH remained almost stationary in the 4% glucose-agar, rose slowly in the 2% glucose-agar, and rose sharply in the 1% glucose-agar. The growth of Histoplasma was inhibited completely at pH 4 and below. When the pH was controlled in mixed cultures, some growth of Histoplasma was obtained. Substitution of maltose for glucose delayed the development of acidity and allowed the appearance of numerous mycelial colonies in the presence of Candida. This growth was arrested as soon as the medium became acid. Four other species which also acidified the Sabouraud's medium effected similar inhibition. It was thus shown that severe and prolonged acidity produced by some yeasts in the sugar-rich Sabouraud's media is alone sufficient to completely inhibit Histoplasma during the standard 4-week incubation of specimens such as sputum.     

A Preliminary Investigation into the Role of Yeasts in the Ensiling Process

The activities of six unidentified yeasts isolated from silage were monitored in a defined environment prepared to simulate a grass silage. Lactic acid and acetic acid contents of the silages were higher when yeasts were included in the inoculum used and the yeasts appeared to be beneficial to preservation. The yeasts also contributed to the small quantities of ethanol which were detected. Where a yeast was used as the sole inoculum the pH of the silage remained high, but lactic acid and acetic acid as well as ethanol were present in the resulting silages.     

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.     

Yeasts associated with algarrobo trees (Prosopis spp.) in northwest Argentina: A preliminary report

Summary Yeasts were isolated from exudates from trees at three sites in northwest Argentina, two between the towns of Amaicha del Valle and Cafayate and one in the Quebrada de Cafayate, a deep river valley north of Cafayate. The majority of the yeasts were identified asCandida famata andRhodotorula graminis, though isolates of other species ofRhodotorula, Candida boidinii, Pichia membranaefaciens, and occasional isolates of other species were obtained. None of the species was the same as those isolated in Crete, from pods of the carob (European algarrobo). Of 96 cultures investigated, 26 utilized methanol as sole carbon source. The frequency of isolation of methylotrophic yeasts from this habitat may prove to be of considerable scientific and technological interest.This paper is dedicated to Professor Herman Jan Phaff in honor of his 50 years of active research which still continues.     

Effect of L-amino acids on Mucor rouxii dimorphism.

Mucor rouxii organisms growing aerobically and exponentially on a well-defined minimal medium are able to differentiate as yeasts or as mycelia, depending on the amino acid as the nitrogen source. When certain amino acids were used as the nitrogen source, spores differentiated only as hyphae, whereas other amino acids gave rise to other morphological forms having different ratios of yeasts to hyphae. In both hyphal and yeast cultures, an aerobic metabolism was predominant, as shown by determining several metabolic parameters such as oxygen tension, glucose consumption, ethanol production, and CO2 release. A complete conversion of yeasts to hyphae was obtained by the appropriate change in the amino acid used as nitrogen source. By preparing spheroplasts from mycelial cultures and transferring them to media with amino acids that induce yeast formation, a 50% yield in the reverse transformation was achieved. A correlation between the change in pH of the medium and cell morphology was observed in different growth conditions. Decrease in the pH of the medium preceded the appearance of hyphae. Also, when the initial pH of the medium was increased, aspartate-containing cultures developed mainly as mycelia, instead of yeasts, with a corresponding decrease in the final pH.     

Isolation of proteolytic psychrotrophic yeasts from fresh raw seafoods

A total of 103 cultures of yeasts were isolated from seven kinds of fresh raw seafoods. The isolates comprised six genera, Candida, Cryptococcus, Debaryomyces, Rhodotorula, Sterigmatomyces and Trichosporon , and included 21 different species. All the isolates were psychrotrophic yeasts. Proteolytic activities of 50 psychrotrophic strains were studied by use of skim milk within the temperature range of 0–42°C. All the strains showed various degrees of proteolysis. In particular, Candida lipolytica. Trichosporon pullulans and Candida scottii were active species at low temperatures. Sensory spoilage due to the proteolytic yeasts were observed in mackerel homogenates stored at 10°C. C. lipolytica -inoculated homogenates caused spoilage with ammoniacal odours after 1 week of storage. Values of total volatile basic nitrogen at 10°C were highest with C. lipolytica among 35 strains tested, followed by Tr. pullans. Trichosporon cutaneum, C. scottii, Rhodotorula glutinis and Cryptococcus luteolus. Proteolytic psychrotrophic yeasts were widely distributed in raw seafoods.     

Microbiology of sayur asin fermentation

Summary Sayur asin is a fermented mustard cabbage product of Indonesia. The cabbage is fermented naturally in the presence of brined water taken from boiled rice. Fermentation was characterized by a sequential growth of the lactic acid bacteria, Leuconostoc mesenteroides, Lactobacillus confusus, Lactobacillus curvatus, Pediococcus pentosaceus, and Lactobacillus plantarum. Starch degrading species of Bacillus, Staphylococcus and Corynebacterium exhibited limited growth during the first day of fermentation. The yeasts, Candida sake and Candida guilliermondii contributed to the fermentation. Lactic acid, acetic acid, succinic acid, ethanol and glycerol were products of fermentation. Glucose, generated by the degradation of rice starch and maltose, was metabolized by the species that grew.     

Process for the Stereoselective Biotransformation of Acetoacetic Acid Esters Using Yeasts

A process for the stereospecific reduction of acetoacetic acid esters to the 3-(S)-hydroxy-butanoic acid esters by the yeasts Saccharomyces cerevisiae and Candida utilis grown on glucose and ethanol media was developed. A continuous single stage steady state production system was found to be superior to pulse-, batch- and fed-batch systems in terms of optical product purity, biomass concentration and production rates.

Optical purity of 3-(S)-hydroxybutanoic acid esters produced with Saccharomyces cerevisiae and Candida utilis was dependent on pH. A maximal optical purity was obtained at pH2.2 from S. cerevisiae growing on ethanol medium. The specific product formation rate of the chemostat cultures was 0.02…0.05 gg^?1 h^?1. C. utilis was more productive than S. cerevisiae but it reconsumed the product under carbon limited growth conditions.     

赤霉烯酮的微生物还原

用Rhodtorula sp.Saccharomyces sp.Arthorbacter sp.和Candida sp．四属20株菌对由Fusarium graminearum 产生的类雌激素——赤霉烯酮(Zearalenone)的还原转化进行了研究。实验结果证明,Rhodotorula sp和 Arthrobacter sp．的还原产物主要是α-赤霉烯醇(α-Zearalenol,经HPLC鉴定含量分别为96％和84％)。Saccharomyces sp．和Candidasp.的主要还原产物是β-赤霉烯醇(β-Zearalenol,经HPLC鉴定含量分别为91％和92％)。产物均经HPLC、13C—NMR和MS鉴定确证。     

Polygalacturonase enzyme production from bacterial isolated from raw milk and green and black olives

Forty microbial strains isolated from raw milk samples and black and green olives were grown in MP5 (mineral pectin 5) medium containing 0.5% lemon pectin. All strains synthesized an extracellular polygalacturonase. Rhodotorula sp. ONRh9 (0.44 U x mL(-1)) and Leuconostoc sp. LLn1 (0.16 U x mL(-1)), which had a more active polygalacturonase in MP5 medium, were studied in MAPG5 medium containing polygalacturonic acid. Highest biomass and polygalacturonase production by these two strains were observed for polygalacturonic acid concentrations of 10 g x L(-1) (Rhodotorula sp. ONRh9) and 5 g x L(-1) (Leuconostoc sp. LLn1) and for initial pH values of 6 (Rhodotorula sp. ONRh9) and 5.5 (Leuconostoc sp. LLn1). The two strains grown in fermenters in MAPG5 medium generated the following results: with controlled initial pH, Rhodotorula sp. produced maximum biomass (DO) and polygalacturonase (PG) after 20 h (DO, 3.86; PG, 0.24 U x mL(-1)) of growth, and this level was sustained until the end of the culture; Leuconostoc sp. LLn1 synthesized more cells and polygalacturonase between 4 h (DO, 1.80; PG, 0.17 U x mL(-1)) and 24 h (DO, 3.90; PG, 0.27 U x mL(-1)) of culture. With uncontrolled initial pH, the cultures produced maximum biomass and polygalacturonase after 20 h (DO, 3.30; PG, 0.26 U x mL(-1)) for Rhodotorula sp. ONRh9 and 10 h (DO, 2.84; PG, 0.17 U x mL(-1)) for Leuconostoc sp. LLn1.     

A study of sulfite-tolerant yeasts from comminuted lamb products

The sulfite tolerance of meat yeasts was shown to be determined by pH, sulfite concentration, substrate availability, and the composition of the preincubation medium. Acetaldehyde production by Candida norvegica was sulfite-induced and occurred during the exponential growth phase in sulfited (500 micrograms SO2 ml-1) lab lemco glucose broth cultures buffered at pH 5, 6, or 7. Growth at pH 4, however, was inhibited by sulfite. Acetaldehyde production occurred in sulfited medium containing fructose or ethanol but not lactate nor a range of other assimilable substrates. A non-acetaldehyde-producing yeast, Candida vini, grew in sulfited (500 micrograms SO2 ml-1) lab lemco broth containing glucose or lactate buffered at pH 6 or 7 but not at pH 4 or 5.     

Characteristics of Yeasts Isolated from Pacific Crab Meat

A total of 202 cultures of yeasts were isolated and characterized from king crab and Dungeness crab meat. A yeastlike organism, resembling Aureobasidium pullulans, and 15 different species distributed among the genera Rhodotorula, Cryptococcus, Torulopsis, Candida, and Trichosporon were represented. Nine of the species grew at 5 C or lower. Although two of the species grew at 37 C, none of the isolates had the characteristics of pathogenic species. Members of the Cryptococcus and Candida failed to grow at 37 C. Furthermore, species of the former genus were not pathogenic to mice. The pigmentation of the Rhodotorula cultures decreased in intensity as the incubation temperature was decreased. Biochemical activities of the different species were studied by use of triglycerides, lecithin, and proteins (casein, gelatin, and crab-meat protein) as substrates. Eight of the species could attack triglycerides; eight, lecithin; five, gelatin; one, casein; and one, crab protein. An organism, tentatively identified as Trichosporon sp., was very active in attacking each of the substrates tested and grew well at 0.5 C.     

Simultaneous production of 2,3-butanediol, ethanol and hydrogen with a Klebsiella sp. strain isolated from sewage sludge

A Klebsiella sp. HE1 strain isolated from hydrogen-producing sewage sludge was examined for its ability to produce H(2) and other valuable soluble metabolites (e.g., ethanol and 2,3-butanediol) from sucrose-based medium. The effect of pH and carbon substrate concentration on the production of soluble and gaseous products was investigated. The major soluble metabolite produced from Klebsiella sp. HE1 was 2,3-butanediol, accounting for over 42-58% of soluble microbial products (SMP) and its production efficiency enhanced after increasing the initial culture pH to 7.3 (without pH control). The HE1 strain also produced ethanol (contributing to 29-42% of total SMP) and a small amount of lactic acid and acetic acid. The gaseous products consisted of H(2) (25-36%) and CO(2) (64-75%). The optimal cumulative hydrogen production (2.7 l) and hydrogen yield (0.92molH(2)molsucrose(-1)) were obtained at an initial sucrose concentration of 30gCODl(-1) (i.e., 26.7gl(-1)), which also led to the highest production rate for H(2) (3.26mmolh(-1)l(-1)), ethanol (6.75mmolh(-1)l(-1)) and 2,3-butanediol (7.14mmolh(-1)l(-1)). The highest yield for H(2), ethanol and 2,3-butanediol was 0.92, 0.81 and 0.59molmol-sucrose(-1), respectively. As for the overall energy production performance, the highest energy generation rate was 27.7kJh(-1)l(-1) and the best energy yield was 2.45kJmolsucrose(-1), which was obtained at a sucrose concentration of 30 and 20gCODl(-1), respectively.     

Occurrence and Growth of Yeasts in Yogurts

Yogurts purchased from retail outlets were examined for the presence of yeasts by being plated onto oxytetracycline malt extract agar. Of the 128 samples examined, 45% exhibited yeast counts above 10³ cells per g. A total of 73 yeast strains were isolated and identified as belonging to the genera Torulopsis, Kluyveromyces, Saccharomyces, Candida, Rhodotorula, Pichia, Debaryomyces, and Sporobolomyces. Torulopsis candida and Kluyveromyces fragilis were the most frequently isolated species, followed by Saccharomyces cerevisiae, Rhodotorula rubra, Kluyveromyces lactis, and Torulopsis versatilis. The growth of yeasts in yogurts was related to the ability of the yeasts to grow at refrigeration temperatures, to ferment lactose and sucrose, and to hydrolyze milk casein. Most yeast isolates grew in the presence of 100 μg of sorbate and benzoate preservatives per ml. Higher yeast counts from yogurts were obtained when the yogurts were plated onto oxytetracycline malt extract agar than when they were plated onto acidified malt extract agar.     

In vitro propagation of Gladiolus hybridus Hort.: Synergistic effect of heat shock and sucrose on morphogenesis

Three cultivars (cvs.) of Gladiolus hybridus Hort., namely ‘Her Majesty’, ‘Aldebaran’ and ‘Bright Eye’ were successfully micropropagated. The cultures were established using intact cormels or segments of cormels and inflorescence axes on Murashige and Skoog (1962; MS) medium. The response depended on media supplements; both callus formation or direct induction of shoot buds was observed. Shoot differentiation from callus could be obtained on MS medium containing 1.0 μM BA (6–benzyladenine) and 10.0 μM NAA (α-naphthalene acetic acid) in all three cultivars. The same could be achieved by giving a heat shock (HS; 50 °C, 1h) to callus cultures (in case of ‘Her Majesty’ and ‘Aldebaran’ only) maintained on the basal medium. In these two cultivars, high sucrose concentration (0.232, 0.290 or 0.348 M) also favoured growth and proliferation of shoot cultures on a plant growth regulator-free medium at 20 °C in comparison to the cultures kept at 25 °C. On the other hand, shoot cultures maintained on the basal medium at 25 °C containing normal (0.058 M, i.e., 2.0%, w/v) sucrose concentration responded similar to those maintained at 20 °C on a high sucrose medium; reduced response was observed on normal sucrose containing medium at 20 °C. Heat shock enhanced shoot proliferation in the cultures maintained on basal medium, but induced prolific rooting in shoot cultures, within 5 days of HS, on high sucrose (optimum 0.232 M) medium. While the number of roots increased at higher sucrose concentrations in the medium in case of cvs. ‘Her Majesty’ and ‘Aldebaran’, the same was found to be independent of sucrose concentration in cv. ‘Bright Eye’. Generally the rooted plants produced on high sucrose (0.232 M) medium in comparison to medium with normal sucrose concentration showed better survival (ca. 90% as against 40%) in the soil. This revised version was published online in June 2006 with corrections to the Cover Date.     

Search for yeast producers of brassylic and sebacic fatty acids

Yeast cultures belonging to the genera Candida, Torulopsis, Saccharomyces, Debaryomyces, Hansenula, Pichia, and Yarrowia, capable of synthesizing brassylic and sebacic fatty acids, were screened. Overall, about 200 cultures grown in media containing decane or tridecane as a sole source of carbon were tested. On the medium with tridecane, yeasts synthesized insignificant amounts of brassylic acid. Sebacic acid was produced more intensively in the medium with n-decane. The culture Candida tropicalis, displaying the highest ability to synthesize sebacic acid, was selected.     

Growth ofCandida utilis on a mixture of monosaccharides,acetic acid and ethanol as a model of waste sulphite liquor

Candida utilis cultivated under batch conditions in a synthetic medium with a mixture of different carbon sources utilized first D-glucose and then D-galactose, D-mannose, D-xylose, L-arabinose, ethanol and acetic acid. The effect of acetic acid was primarily a function of pH and the physiological state of the inoculum. At pH 4.5, acetic acid at a concentration of 1 g/l increased the specific growth rate, reduced time of cultivation and increased yield of the yeast dry weight. The yield from acetic acid was 61%. In the presence of a higher content of acetic acid (3--6 g/l) the yield was only 18--26%. The yield calculated only from monosaccharides increased but the yield with respect to total carbon sources was lower. The specific growth rate decreased as well. The addition of ethanol also resulted in an increase of the production and yield of the yeast dry weight but the cultivation time was prolonged. The simultaneous utilization of carbon sources of the studied mixture modelling a sulphite fermentation medium with ethanol is advantageous. However, due to physiology of the yeast, it is most suitable to cultivate a strain adapted to utilizable carbon sources in a continuous way, in the presence of their limiting concentrations in the cultivation medium.