Quantification of glycosidase activities in selected yeasts and lactic acid bacteria

Using a model system, the activities of α-L-arabinofuranosidase, β-glucosidase, and α-L-rhamonopyranosidase were determined in 32 strains of yeasts belonging to the genera Aureobasidium, Candida, Cryptococcus, Hanseniaspora, Hansenula, Kloeckera, Metschnikowia, Pichia, Saccharomyces, Torulaspora and Brettanomyces (10 strains); and seven strains of the bacterium Leuconostoc oenos. Only one Saccharomyces strain exhibited β-glucosidase activity, but several non-Saccharomyces yeast species showed activity of this enzyme. Aureobasidium pullulans hydrolyzed α-L-arabinofuranoside, β-glucoside, and α-L-rhamnopyranoside. Eight Brettanomyces strains had β-glucosidase activity. Location of enzyme activity was determined for those species with enzymatic activity. The majority of β-glucosidase activity was located in the whole cell fraction, with smaller amounts found in permeabilized cells and released into the growth medium. Aureobasidium pullulans hydrolyzed glycosides found in grapes. Received 02 February 1999/ Accepted in revised form 26 June 1999     

Assessment of phylloplane yeasts on selected Mediterranean plants by FISH with group- and species-specific oligonucleotide probes

A previous culture-dependent survey of phylloplane yeasts from selected Mediterranean plants showed that a few species were present in high densities in almost all leaf samples, regardless of the plant type, location or sampling season. However, a few species appeared to be restricted to Cistus albidus leaves, namely Cryptococcus cistialbidi . Here, we describe a culture-independent FISH assay to detect and quantify whole yeast cells in leaf washings. After optimization, the technique was used to check the apparent association between C. albidus leaves and C. cistialbidi and the abundance and ubiquity of other basidiomycetous yeast species such as Erythrobasidium hasegawianum and Sporobolomyces spp. in leaf samples from this and other neighboring plants ( Acer monspessulanum and Quercus faginea ). No yeast cells were detected in Pistacia lentiscus leaf samples. We were also able to demonstrate that three phylloplane yeasts ( C. cistialbidi, E. hasegawianum and Sporobolomyces spp.) appeared to be log-normally distributed among individual C. albidus leaves. The log-normal distribution has important implications for the quantification of phylloplane yeasts based on the washing and plating of bulk leaf samples, which will tend to overestimate the size of the respective populations and become an error source in yeast surveys or related biocontrol studies.     

Salt stress and plasma-membrane fluidity in selected extremophilic yeasts and yeast-like fungi

We have investigated changes in plasma-membrane fluidity in relation to NaCl concentrations in yeasts and yeast-like fungi that were isolated from either subglacial ice or hypersaline waters. In both of these natural environments, these organisms are exposed to low water activity, due to either high NaCl concentrations or low temperatures. Our data indicate that the fluidity of the plasma membrane can be used as an indicator of fitness for survival in extreme environments. Fungi that can survive in such extreme environments, such as Hortaea werneckii in the hypersaline waters of salterns, and Cryptococcus liquefaciens in subglacial environments, showed similar profiles of plasma-membrane fluidity in response to raised salinity. The same was seen for ubiquitous fungi, which are generally adapted for different types of stress, such as Aureobasidium pullulans and Rhodotorula mucilaginosa. Representatives of both of these groups modulated their plasma-membrane fluidity differently. When salinity exceeded their optimal range, the ubiquitous stress-tolerant species (A. pullulans, Rh. mucilaginosa) showed increased plasma-membrane fluidity, whereas in the dominant extremophiles (H. werneckii, Cr. liquefaciens), it decreased. On the other hand, the plasma membranes of the fungi with a narrow ecological amplitude (Arctic A. pullulans and Rhodosporium diobovatum) showed different responses.     

Xylose fermentation by yeasts

Summary Xylose reductase from the xylose-fermenting yeastPichia stipitis was purified to electrophoretic homogeneity via ion-exchange, gel and affinity chromatography. At physiological pH values the thermodynamic equilibrium constant was determined to be 0.575x10¹⁰ (l·mol^-1). Product inhibiton studies are reported which clearly show that the kinetic mechanism of the xylose reductase is ordered-bi-bi with isomerisation of a stable enzyme form.     

Xylose fermentation by yeasts

Summary Utilization and fermentation of xylose by the yeasts Pachysolen tannophilus I fGB 0101 and Pichia stipitis 5773 to 5776 under aerobic and anaerobic conditions are investigated. Pa. tannophilus requires biotin and thiamine for growth, whereas Pi. stipitis does not, and growth of both yeasts is stimulated by yeast extract. Pi. stipitis converts xylose (30 g/l) to ethanol under anaerobic conditions with high yields of 0,40 and it produces only low amounts of xylitol. The yield coefficient is further increased at lower xylose concentrations.Publication Nr. 2 of this series: Eur. J. Appl. Microbiol. Biotechnol. (1983) 17, 287–291.     

Fingerprinting of yeasts at the strain level by differential sensitivity responses to a panel of selected killer toxins

We used differential sensitivities to a panel of twenty-five cell-free crude killer toxins to fingerprint forty-four Saccharomyces cerevisiae strains of different origin and all taxonomically certified by nDNA-nDNA reassociation. Cluster analysis of numerical data obtained by different growth inhibition areas observed in Petri dishes allowed the complete and reproducible discrimination of all S. cerevisiae strains.     

Photo-sensitized Isomerization of Retinal by Dyes

THE initial reaction following absorption of light in the retina is the isomerization of the 11-cis retinal chromophore of the visual pigment¹. Isolated 11cis retinal will undergo the same isomerization to the all-trans form when excited by light of wavelength shorter than about 450 nm and this reaction can be sensitized to light of longer wavelengths by the addition of trace amounts of iodine to the solution².     

Long-chain fatty acid composition of selected genera of yeasts belonging to the Endomycetales

The cellular long-chain fatty acids of 36 strains representing 18 genera of the Saccharomycetaceae, Endomycetaceae, Metchnikowiaceae, Saccharomycodaceae, Schizosaccharomycetaceae and Dipodascaceae were extracted and analyzed as methyl esters by gas chromatography. On the basis of their fatty acid content the set of strains was divided into 6 groups, coinciding with the above families. The members of the Saccharomycetaceae (group I) had a high percentage of oleic acid while the strains classified under the Endomycetaceae (group II) and Metchnikowiaceae (group III) were characterized by oleic acid and linoleic acid as major fatty acids. The Saccharomycodaceae (group IV) had the highest percentage of palmitoleic acid. The Schizosaccharomycetaceae (group V) had the highest percentage of oleic acid, while the Dipodascaceae (group VI) were characterized by a high percentage of linoleic acid.     

Long-chain fatty acid composition of selected genera of yeasts belonging to the Endomycetales

The cellular long-chain fatty acids of 36 strains representing 18 genera of the Saccharomycetaceae, Endomycetaceae, Metchnikowiaceae, Saccharomycodaceae, Schizosaccharomycetaceae and Dipodascaceae were extracted and analyzed as methyl esters by gas chromatography. On the basis of their fatty acid content the set of strains was divided into 6 groups, coinciding with the above families. The members of the Saccharomycetaceae (group I) had a high percentage of oleic acid while the strains classified under the Endomycetaceae (group II) and Metchnikowiaceae (group III) were characterized by oleic acid and linoleic acid as major fatty acids. The Saccharomycodaceae (group IV) had the highest percentage of palmitoleic acid. The Schizosaccharomycetaceae (group V) had the highest percentage of oleic acid, while the Dipodascaceae (group VI) were characterized by a high percentage of linoleic acid.This article originally appeared in an incorrect form in Antonie van Leeuwenhoek, Vol. 52, No. 1.     

Sulfite formation by wine yeasts

The enzyme catalyzing the reduction of sulfite by reduced benzyl viologen (BVH) was partially purified and characterized from two strains of wine yeasts, a sulfite-producing strain and a non-producing strain.Both enzymes showed corresponding features in pH-optima, optima of buffer and benzyl viologen concentrations.The enzymes did not catalyze the reduction of nitrite by reduced viologen dyes, but the reduction of sulfite was uncompetitively inhibited by nitrite. Compounds of sulfur metabolism such as sulfate, thiosulfate, cysteine, serine and methionine did not influence the activity of either of the enzymes. The main differences between the two enzymes exist in the specific activities in crude extracts, the K _m -values for sulfite, substrate inhibition rates, and localization in different fractions during (NH₄)₂SO₄ precipitation. The specific activity in crude extracts of the sulfite-producing strain (0.052 moles S^2- x min^-1 x mg^-1) was about three fold higher than that of the non-producing strain (0.0179 moles S^2- x min^-1 x mg^-1). On the other hand the sulfite-producing strain had a higher K _m -value for sulfite (2×10^-3 M) and was more strongly inhibited by the substrate than the non-producing strain (6×10^-3 M).     

Sulfite formation by wine yeasts

Five different strains of wine yeasts were investigated with respect to active uptake of [³⁵S] sulfate and its regulation by methionine. Considerable differences exist between low and high sulfite-producing strains in the initial velocity of sulfate uptake. Further differences were established in repression of sulfate permease by l-methionine, most evident in a total lack of repression in one of the high sulfite producers. These findings explain in part variable sulfite and sulfide formation.List of Abbreviations CCMP carbonylcyanide-p-trifluormethoxyphenylhydrazone     

Production of biotin by yeasts

A quantitative screening procedure for biotin and biotin vitamer production was conducted on 129 yeast strains able to grow in a biotin-free medium. Production of biotin and biotin vitamers varied considerably from strain to strain even within a species. The best producers of biotin were strains of Sporobolomyces roseus and Rhodotorula rubra whilst strains of Rhodotorula rubra and Yarrowia lipolytica produced the largest amounts of vitamers.     

Sulfite formation by wine yeasts

Four strains of wine yeasts of two different species (Saccharomyces cerevisiae var. ellipsoideus and S. bayanus) were investigated with respect to regulation of NADPH- and benzyl viologen-dependent sulfite reductases by various sulfur sources. The enzyme activity was followed over a growth period of 96 h. The low sulfite-producing strains showed an increased biosynthesis of NADPH-dependent sulfite reductase during the exponential growth phase in the presence of sulfate, sulfite and djencolic-acid. This increase was not observed in the high sulfite-producing strains. Methionine and cysteine prevented this derepression. At the end of the exponential growth phase, enzyme biosynthesis was repressed again, presumably by sulfur-containing amino acids which were produced during growth. The regulatory influence of the various sulfur sources on benzyl viologen dependent sulfitereductase activity is obviously much weaker.Abbreviation BV benzyl viologen