Selenium tolerance of yeasts

Selenium tolerance of yeasts widely varies: the growth of some yeasts can be inhibited by a selenium concentration as low as 10(-4) M, whereas others can grow in the presence of 10(-1) M selenium. Homogeneous yeast taxa are characterized by a certain level of selenium tolerance, and heterogeneous taxa show a variable level of tolerance to selenium. In general, ascomycetous yeasts are more tolerant to selenium than basidiomycetous yeasts. Among the ascomycetous yeasts, the genera Dekkera and Schizosaccharomyces exhibited the lowest and the species Candida maltosa, Hanseniaspora valbyensis, Kluyveromyces marxianus, and Yarrowia lipolytica the highest tolerance to selenium. Among the basidiomycetous yeasts, the genera Bullera, Cryptococcus, and Holtermannia showed the lowest and the species Cryptococcus curvatus, Cr. humicola, and Trichosporon spp. the highest tolerance to selenium. The selenium tolerance of yeasts depends on the composition of the growth medium, in particular, on the presence of sulfate, sulfur-containing amino acids, and glutamine in the medium.     

Silver tolerance and accumulation in yeasts

Summary Debaryomyces hansenii (NCYC 459 and strain 75-21),Candida albicans (3153A),Saccharomyces cerevisiae (X2180-1B),Rhodotorula rubra (NCYC 797) andAureobasidium pullulans (IMI 45533 and ATCC 42371) were grown on solid medium supplemented with varying concentrations of AgNO₃. Although Ag⁺ is highly toxic towards yeasts, growth on solid media was still possible at Ag concentrations of 1–2 mM. Further subculture on higher Ag concentrations (up to 5 mM) resulted in elevated tolerance. The extent of Ag tolerance depended on whether Ag-containing plates were exposed to light prior to inoculation since light-mediated reduction of Ag⁺ to Ag⁰ resulted in the production of a less toxic silver species. Experimental organisms exhibited blackening of colonies and the surrounding agar during growth on AgNO₃-containing medium especially at the highest Ag concentrations tested. All organisms accumulated Ag from the medium; electron microscopy revealed that silver was deposited as electron-dense granules in and around cell walls and in the external medium. X-ray microprobe analysis indicated that these granules were metallic Ag⁰ although AgCl was also present in some organisms. Volatile and non-volatile reducing compounds were produced by several test organisms which presumably effected Ag⁺ reduction to Ag⁰.     

工业酵母抗逆机理研究进展

工业酵母利用木质纤维素等生物质资源发酵生产醇、酮、醛、酸等各种化合物,是解决人类面临的不可再生资源和能源危机的重要途径,这激发了人们对木质纤维素水解液为原料和环保节能型浓醪发酵技术的极度关注。然而高浓度底物、产物、渗透压、木质纤维素水解液中抑制性物质、发酵过程温度的提高均会抑制微生物生长代谢及发酵性能,这是发酵行业"瓶颈"问题。本文简述了渗透压、温度及抑制性物质对酵母细胞生长的危害,并从胞内稳态平衡、分子水平等方面着重叙述工业酵母对渗透压、温度及抑制性物质的抗逆机制研究进展。     

Comparison of ethanol tolerance of free and immobilizedSaccharomyces uvarum yeasts

O₂ consumption and CO₂ production of free and immobilizedSaccharomyces uvarum in the presence of ethanol were compared. The protective effect of immobilization on the yeast ethanol tolerance at 5–20% of ethanol was more evident in CO₂ production than in O₂ consumption. CO₂ production by the yeast immobilized in calcium alginate and calcium pectate gel beads was approximately 2.5-times higher than by the free yeast at 5 and 10% of ethanol. 4-Fold increase of CO₂ production was observed at 15% ethanol. Immobilization in calcium-containing carriers (alginate, pectate) resulted in enhanced activities of yeasts compared to the κ-carrageenan carrier.     

Extremophilic yeasts: plasma-membrane fluidity as determinant of stress tolerance

Our aim was to investigate the response of selected yeasts and yeast-like fungi from extreme?environments to various temperatures at the level of their plasma membranes, in order to elucidate the connections between their plasma-membrane fluidity (measured by electron paramagnetic resonance spectroscopy - EPR), growth temperature range, stress tolerance, and ecological distribution. Although all studied fungi can be considered mesophilic according to their growth temperature profiles, their plasma-membrane fluidity indicated otherwise. Arctic yeast Rhodosporidium diobovatum could be classified as psychrotolerant?due to its higher average membrane fluidity. Extremely halotolerant black yeast-like fungus Hortaea werneckii isolated from solar salterns, on the other hand, is not adapted to low temperature, which is reflected in the higher average rigidity of its plasma membrane and as a consequence its inability to grow at temperatures lower than 10°C. The plasma membrane of Aureobasidium sp. isolated so far exclusively from an Arctic glacier with its intermediate fluidity and high fluidity variation at different temperatures may indicate the specialization of this yeast-like fungus to the specific glacial environment. Similar behaviour of plasma membrane was detected in the reference yeast, non-extremophilic Saccharomyces cerevisiae. Its membranes of intermediate fluidity and with high fluidity?fluctuation at different temperatures may reflect the specialization of this yeast to mesophilic environments and prevent its colonization of extreme environments. Halotolerant Aureobasidium pullulans from salterns, and Arctic Cryptococcus liquefaciens and Rhodotorula?mucilaginosa with moderately fluctuating plasma membranes of intermediate fluidity are representatives of globally distributed generalistic and stress-tolerant species that can thrive in a variety of environments. Keeping the membranes stable and flexible is one of the necessities for the microorganisms to survive changes in extreme habitats. Our data suggest that plasma-membrane fluidity can be used as an indicator of fitness for survival in the extreme environments. In addition to the average fluidity of plasma membrane, the fluctuation of fluidity is an important determinant of stress tolerance: high absolute fluidity fluctuation is tied to decreased survival. The fluidity and its variation therefore reflect survival strategy and fitness in extreme environments and are good indicators?of the adaptability of microorganisms.     

Inferring ethanol tolerance of Saccharomyces and non-Saccharomyces yeasts by progressive inactivation

The kinetics of cell inactivation in the presence of ethanol at 20, 22.5% and 25% (v/v), was measured by progressive sampling and viable counting, and used as an inference of the ethanol resistance status of five non-Saccharomyces strains and one strain of Saccharomyces cerevisiae. The capacity of standard inocula of the same strains to establish growth at increasing initial ethanol concentrations was employed as a comparison. The effect of various different pre-culture conditions on the ethanol resistance of the 6 strains was analysed by the cell inactivation method and by the cell growth method. Exposing cells to 25% (v/v) ethanol for 4 min enabled the differentiation of the yeasts in terms of their resistance to ethanol. The results suggest that the two methods are generally concordant and that the cell inactivation method can, thus, be used to infer ethanol resistance of yeast strains.     

Determination of iron in saccharomycete yeasts by the dipyridyl method

Iron content in intact yeast cells without their incineration or disintegration can be determined using 2,2'-dipyridyl.     

Self-cloning baker's yeasts that accumulate proline enhance freeze tolerance in doughs

We constructed self-cloning diploid baker's yeast strains by disrupting PUT1, encoding proline oxidase, and replacing the wild-type PRO1, encoding gamma-glutamyl kinase, with a pro1(D154N) or pro1(I150T) allele. The resultant strains accumulated intracellular proline and retained higher-level fermentation abilities in the frozen doughs than the wild-type strain. These results suggest that proline-accumulating baker's yeast is suitable for frozen-dough baking.     

Studies on marine occurring yeasts: Respiration,fermentation and salt tolerance

Summary The effect of various NaCl concentrations on respiration and fermentation rates in cells with or without added glucose as exogenous substrate as well as on respiratory quotients was determined for Debaryomyces hansenii, Saccharomyces cerevisiae, Cryptococcus albidus, and Candida zeylanoides, all yeasts isolated from marine environment. A given strain had about the same respiratory and fermentatory intensity at 0% and 4% NaCl (w/v). A further increase considerably reduced the oxygen uptake or CO₂-evolution. D. hansenii was the most NaCl tolerant yeast tested, giving about 10% activity still at a concentration of 24% NaCl, whether the activities of whole cells or cell homogenates were determined. For S. cerevisiae or Cr. albidus the respiratory activity was reduced to about the same degree at 16% NaCl for whole cells, at 12% NaCl for homogenates of Cr. albidus. A somewhat higher NaCl concentration was evidently tolerated for respiration and fermentation than for growth, very obvious in the case of C. zeylanoides.The minimum values for water activity (a _w) permitting 10% respiration activity were higher when produced by electrolytes (NaCl, KCl, or Na₂SO₄), lower when due to sugars (metabolizable glucose or non-metabolizable lactose) and lowest when due to glycerol. The a _w per se was evidently not solely decisive for the limitation of respiration activity.Attempts were made to assess an effect of high NaCl concentrations on the glucose uptake.The potassium content was higher in cells of the highly halotolerant D. hansenii than in those of the other yeasts and decreased with the increase in external, consequently in internal, Na⁺ concentration. The decrease in K⁺ content can presumably only proceed to a certain extent, below which the ability for growth and respiration was lost.     

Temperature profiles of growth and ethanol tolerance of the xylose-fermenting yeasts Candida shehatae and Pichia stipitis

Summary The effect of different ethanol concentrations on the growth of Candida shehatae and Pichia stipitis with xylose as substrate was evaluated in a temperature gradient incubator. The upper limit of the temperature profiles of ethanol tolerance of both yeast strains were similar, although P. stipitis appeared to have a slightly higher ethanol tolerance in the higher temperature range. An increase in the ethanol concentration severely depressed the maximum growth temperature, and also increased the minimum growth temperature slightly. The ethanol tolerance limit of 46–48 g·l^-1 occurred within a narrow temperature plateau of 11 to 22° C. The low ethanol tolerance of these pentose fermenting yeasts is detrimental for commercial ethanol production from hemicellulose hydrolysates.     

三种食用植物酵素中酵母菌的分离鉴定及耐受性试验

从自然发酵的火龙果酵素、青梅酵素和葡萄酵素中筛选酵母菌,对筛选到的菌株进行鉴定及耐受性试验。从两种火龙果酵素中筛选到3株菌:P_1、P_2和P_3,从青梅酵素中筛选到2株菌:PM_1和PM_2,从葡萄酵素中筛选到2株菌:G_1和G_2;分子生物学鉴定结果显示:P_3、PM_1、PM_2和G_2为鲁氏接合酵母,P_1为酿酒酵母,P_2和G_1为丘陵假丝酵母。耐受性试验结果表明:筛选出的菌株在起始pH为2. 5、3. 0、3. 5以及初始葡萄糖含量为300、400、600、750 g/L的培养基上均可生长,其中P_2、PM_1和G_1在培养基起始pH为2. 0时可生长;P_2在培养基起始pH为1. 5时可生长;筛选出的菌株均具有耐低pH和耐高糖的特性。     

Creation of a novel peptide endowing yeasts with acid tolerance using yeast cell-surface engineering

The cell wall of Saccharomyces cerevisiae plays an essential role in the biophysical characteristics of the cell surface. The modification of the cell wall property is an important factor for cellular adaptation to a stressful environment. In this study, we randomly modified the cell wall by displaying combinatorial random peptides on the yeast cell surface, and by screening, we successfully obtained a novel peptide, Scr35, that endowed yeasts with acid tolerance. The yeast, surface-modified by Scr35, was able to grow well under acidic condition and low glucose condition and showed high glucose uptake activity. However, the growth of the modified yeast became inferior as extracellular pH became higher. This inferiority was rescued by decreasing glucose concentration in a medium. Our results suggest that the optimum pH of a medium becomes low when the newly created Scr35 affects glucose uptake activity through cell-surface modification. Therefore, such artificial modification of the cell surface has a great potential as a useful tool for breeding acid-tolerant yeasts for industrial applications of S. cerevisiae as a biocatalyst.