Urease and extracellular nucleases of yeasts

Fifty-five representatives of yeast genera and 745 strains of large genera were tested with respect to the production of urease, extracellular RNAase and DNAase. The results made it possible to set up eight taxonomical classes of the yeasts. The taxonomical significance of the classification of these 8 classes is discussed.     

Yeasts Isolated from Neotropical Wood-Boring Beetles in SE Peru

Some temperate wood-boring cerambycid beetles harbor intracellular gut yeasts believed to augment host nutrition, but species belonging to the subfamily Lamiinae are thought to lack endosymbionts. Almost 49 percent of Neotropical cerambycid species are lamiines, therefore, comparatively few rain forest species would be expected to host symbiotic gut yeasts. This study reports the isolation of gut yeasts from closely related Neotropical lamiines. We investigated species that feed on trees in the Brazil nut family (Lecythidaceae), because host plant associations are relatively well known. Our objectives were to determine if gut yeasts were present and, if possible, infer their mode of transmission. We collected and dissected 18 beetle specimens from three tree species, including 17 cerambycids and one curculionid. Every insect specimen yielded a gut yeast. DNA sequence libraries were used for a rapid identification of the yeasts and their larval hosts. The cerambycids included five lamiine species and one cerambycine. Six ascomycete yeasts were isolated from their guts; we found no evidence of strict vertical transmission. Larval gut yeasts were genetically similar to yeasts previously isolated from insects associated with wood or fungi, implying potential habitat specificity. The yeasts have not yet been localized, and potential function is not known, but they may contribute to rapid nutrient cycling or serve as the first line of defense against plant toxins.     

Raman Spectroscopy and Chemometrics for Identification and Strain Discrimination of the Wine Spoilage Yeasts Saccharomyces cerevisiae,Zygosaccharomyces bailii,and Brettanomyces bruxellensis

The yeasts Zygosaccharomyces bailii, Dekkera bruxellensis (anamorph, Brettanomyces bruxellensis), and Saccharomyces cerevisiae are the major spoilage agents of finished wine. A novel method using Raman spectroscopy in combination with a chemometric classification tool has been developed for the identification of these yeast species and for strain discrimination of these yeasts. Raman spectra were collected for six strains of each of the yeasts Z. bailii, B. bruxellensis, and S. cerevisiae. The yeasts were classified with high sensitivity at the species level: 93.8% for Z. bailii, 92.3% for B. bruxellensis, and 98.6% for S. cerevisiae. Furthermore, we have demonstrated that it is possible to discriminate between strains of these species. These yeasts were classified at the strain level with an overall accuracy of 81.8%.     

Application of anhydrobiosis and dehydration of yeasts for non-conventional biotechnological goals

Dehydration of yeast cells causes them to enter a state of anhydrobiosis in which their metabolism is temporarily and reversibly suspended. This unique state among organisms is currently used in the production of active dry yeasts, mainly used in baking and winemaking. In recent decades non-conventional applications of yeast dehydration have been proposed for various modern biotechnologies. This mini-review briefly summarises current information on the application of dry yeasts in traditional and innovative fields. It has been shown that dry yeast preparations can be used for the efficient protection, purification and bioremediation of the environment from heavy metals. The high sorption activity of dehydrated yeasts can be used as an interesting tool in winemaking due to their effects on quality and taste. Dry yeasts are also used in agricultural animal feed. Another interesting application of yeast dehydration is as an additional stage in new methods for the stable immobilisation of microorganisms, especially in cases when biotechnologically important strains have no affinity with the carrier. Such immobilisation methods also provide a new approach for the successful conservation of yeast strains that are very sensitive to dehydration. In addition, the application of dehydration procedures opens up new possibilities for the use of yeast as a model system. Separate sections of this review also discuss possible uses of dry yeasts in biocontrol, bioprotection and biotransformations, in analytical methods as well as in some other areas.     

Mannose-containing polysaccharides of yeasts

The known species of yeasts produce at lest 180 different polysaccharides having mannose as a major constituent. The type of mannan formed is characteristic of the yeast species, and can be used as an aid in identification and classification. The immunological characteristics of yeasts depend largely upon the nature of the mannans. Some of the yeast mannans can be produced in relatively high yields, and have been suggested as replacements for plant and bacterial gums as thickeners, dispersing agents, and similar applications. However, more significant uses, based on the finer differences in their chemical structures and the specific reactions they undergo, may result from a closer study of these polymers.     

Utilization of mixtures of carbon compounds by yeasts

As a tool for the classification of yeasts a method is presented for determining the aerobic utilization of each of 28 carbon compounds distributed over 4 mixtures containing respectively 10, 7, 7 and 4 of these compounds. Paper chromatography is used to follow the eventual disappearance of each individual carbon compound from the medium. This method is compared to the well-known standard method where a growth response to singly offered compounds in liquid medium is used for biochemical characterization. The proposed method has two advantages: results are obtained more rapidly and with less work, and the paper-chromatographic method may reveal a time sequence of utilization. Both the new method and the established standard method were applied to eight yeasts. Although the backgrounds are different, the experimental results were similar. The method presented is suitable for the differentiation of species as well as of strains within one species. It should be considered for use in taxonomy.     

Reconsideration of the role of yeasts associated with Chrysoperla green lacewings

As larvae, lacewings in the genus Chrysoperla (Neuroptera: Chrysopidae: Chrysopinae: Chrysopini) are predators of aphids and other soft-bodied insects. Adult Chrysoperla, however, are not predacious and feed on pollen, nectar, and honeydew. Earlier studies observed that Chrysoperla adults house yeasts in their crops, and based on the results of a sorbic acid yeast suppression experiment, concluded that the yeasts may supplement amino acids missing in the largely carbohydrate diet. In the current study, attempts to cure adult Chrysoperla comanche (Banks) and Chrysoperla carnea (Stephens) of yeasts using Hagen et al.’s [Bull. Lab. Entomol. Agric. Fil Silv. 28 (1970) 113] protocol, as well as several other fungicides and heat treatment, were unsuccessful, thus calling into question earlier conclusions about the yeasts. Based on our findings, we suggest possible methods for producing yeast-free lacewings so that future studies might determine how yeasts contribute to lacewing fitness. The earlier research also suggested that lacewings eclose without their yeast symbionts and must obtain them from the environment. Our data suggest that yeast may be transmitted vertically, from mother to offspring.     

Yeast concentration estimation and prediction with static and dynamic neural network models in batch cultures

The second fermentation is one of the most important steps in Champagne production. For this purpose, yeasts are grown on a wine based medium to adapt their metabolism to ethanol. Several models built with various static and dynamic neural network configurations were investigated. The main objective was to achieve real-time estimation and prediction of yeast concentration during growth. The model selected, based on recurrent neural networks, was first order with respect to the yeast concentration and to the volume of CO₂ released. Temperature and pH were included as model parameters as well. Yeast concentration during growth could thus be estimated with an error lower than 3% (±1.7×10⁶ yeasts/ml). From the measurement of initial yeast population and temperature, it was possible to predict the final yeast concentration (after 21 hours of growth) from the beginning of the growth, with about ±3×10⁶ yeasts/ml accuracy. So a predictive control strategy of this process could be investigated.     

Taxonomic study of four yeast strains assimilating methanol

The new isolated yeasts were very good producers of biomass from methanol. Their taxonomic studies were based on classical classification, GC content of DNA, proton magnetic resonance spectrum of the cell wall mannans and mannan and glucan and chitin contents in the cell walls. The isolates could not be identified with any species described in literature. Considering their special features and some relation to the known species, the isolated yeasts were classified as follows: C-16 as Candida bimundalis var. chlamydospora, C-4 as Candida melinii var. melibiosica, D-3 as Candida silvicola var. melibiosica and M-1 as Torulopsis candida var. nitratophila.     

嗜杀酵母的生物学功能及其应用

嗜杀酵母能够分泌毒素蛋白,杀死敏感酵母。嗜杀酵母对自身分泌的嗜杀毒素具有免疫力。嗜杀酵母的嗜杀特性与两种双链线状RNA(dsRNA)有关,即编码产生毒素蛋白的M-dsRNA和编码自身和M-dsRNA外壳蛋白的L-dsRNA。嗜杀毒素破坏细胞跨膜化学质子梯度,造成ATP和钾离子泄漏,导致细胞死亡。应用嗜杀酵母可避免野生型酵母污染,净化发酵体系,改善发酵产物品质;嗜杀毒素也可作为抵制病原酵母和类酵母微生物的抗真菌剂。     

我国各类基物上的酵母菌分布及其尿素酶活性

本文讨论了2300株由我国各地分离到的酵母菌和类酵母菌与各类基物的关系。共述及23属,占Lodder(1970)系统的半数以上。在十几类基物中酵母菌的分布各有不同,叶子上有大量掷孢酵母,果实上比其它基物更能获得克勒克酵母属(Kloeckera),在咸腌食品中相对地有较多耐高渗透压的酵母如球拟酵母属(Torulopsis)与德巴利酵母属(Debaryomyces)。对部分属的定种工作也已进行,说明某些属的种在我国均有发现而且为数不少。此外还研究了600余株代表各类酵母的尿素酶活性,结果说明担子菌酵母或系统上接近担子菌的酵母都有尿素酶活性,而子囊菌酵母则无。根据作者的观察子囊菌酵母中裂殖酵母属(Schizosaccharomyces)则常常例外地有尿素酶反应,且常有辅酶Q_(10),从而可见裂殖酵母属有其特有的生物学特性,在系统上很可能与其它子囊菌酵母不同。对属于不完全菌的无孢子酵母也进行了尿素酶活性测定,结果表明它们部分与子囊菌有关,部分与担子菌有关。     

甲醇酵母代谢工程研究进展

甲醇酵母由于独特优点被认为是绿色生物制造的潜在宿主。特别是其天然甲醇利用性能有望建立甲醇生物转化路线,拓展生物炼制底物,具有重要经济价值和环保意义。文中综述了代谢工程改造甲醇酵母合成蛋白质和化学品的最新研究进展,并比较了其与模式生物酿酒酵母作为细胞工厂的优缺点。随后,分析了甲醇酵母代谢工程改造面临的挑战,并展望了潜在解决方案。随着基因操作工具开发和细胞代谢阐释,甲醇酵母将在未来绿色生物制造发挥越来越重要的作用。     

Utilization of phenol by hydrocarbon assimilating yeasts

77 Ascomycetous, basidiomycetous as well as imperfect yeast strains of 46 different species and 20 genera were tested for growth with the substrates n-octane, n-hexadecane, and phenol. Of 59 yeast strains with ascomycetous cell wall structure 33 grew on hydrocarbons and 32 on phenol. No yeast strain out of 26 which are unable to use n-alkanes as a source of carbon and energy grew on phenol. In comparison with the latter 32 out of 33 n-hexadecane assimilating yeasts were also capable of using phenol. All n-octane utilizing yeasts of this group also assimilate phenol as a carbon source for growth.The correlation of the hydrocarbon assimilation with the phenol assimilation seems to be not so strong in the basidiomycetous yeasts. 7 out of 18 strains from this group grew on n-hexadecane and 13 on phenol.Furthermore, it could be shown that the use of hydrocarbons and phenol (as well as methanol) is strongly correlated with the coenzyme Q structure of the respective yeast strain.The results are discussed with respect to the particular chemical properties of the substrates used and the fact that coenzyme Q structure is considered to be an important marker of evolutionary relationships among yeasts.     

Electrofusion of fragile mutants of Saccharomyces cerevisiae

This communication presents experimental evidence that intact fragile (osmotic sensitive) yeasts can be electrofused and give viable hybrids. The yield increases with one order of magnitude for electrofusion of intact fragile yeasts with protoplasts of non-fragile ones. The yield of viable hybrids, obtained by electrofusion of protoplasts of fragile and non-fragile yeasts, is one order of magnitude higher than the yield from protoplasts of non-fragile yeasts. The destabilized cell wall and plasma membrane of the mutant yeasts could be a possible explanation for this phenomenon.     

Production,characterization and gene cloning of the extracellular enzymes from the marine-derived yeasts and their potential applications

In this review article, the extracellular enzymes production, their properties and cloning of the genes encoding the enzymes from marine yeasts are overviewed. Several yeast strains which could produce different kinds of extracellular enzymes were selected from the culture collection of marine yeasts available in this laboratory. The strains selected belong to different genera such as Yarrowia, Aureobasidium, Pichia, Metschnikowia and Cryptococcus. The extracellular enzymes include cellulase, alkaline protease, aspartic protease, amylase, inulinase, lipase and phytase, as well as killer toxin. The conditions and media for the enzyme production by the marine yeasts have been optimized and the enzymes have been purified and characterized. Some genes encoding the extracellular enzymes from the marine yeast strains have been cloned, sequenced and expressed. It was found that some properties of the enzymes from the marine yeasts are unique compared to those of the homologous enzymes from terrestrial yeasts and the genes encoding the enzymes in marine yeasts are different from those in terrestrial yeasts. Therefore, it is of very importance to further study the enzymes and their genes from the marine yeasts. This is the first review on the extracellular enzymes and their genes from the marine yeasts.     

Cell surface engineering of yeast for applications in white biotechnology

Cell surface engineering is a promising strategy for the molecular breeding of whole-cell biocatalysts. By using this strategy, yeasts can be constructed by the cell surface display of functional proteins; these yeasts are referred to as arming yeasts. Because reactions using arming yeasts as whole-cell biocatalysts occur on the cell surface, materials that cannot enter the cell can be used as reaction substrates. Numerous arming yeasts have therefore been constructed for a wide range of uses such as biofuel production, synthesis of valuable chemicals, adsorption or degradation of environmental pollutants, recovery of rare metal ions, and biosensors. Here, we review the science of yeast cell surface modification as well as current applications and future opportunities.     

MALDI‐TOF‐MS‐based species identification and typing approaches in medical mycology

MALDI‐TOF MS‐based species identification has found its place in many clinical routine diagnostic laboratories over the past years. Several well‐established commercial systems exist and these allow precise analyses not only among bacteria, but also among clinically important yeasts. This methodology shows higher precision than biochemical and microscopic methods at significantly reduced turnaround times. Furthermore, the differentiation of different filamentous fungi including most dermatophytes and zygomycetes has been established. The direct identification of yeasts from blood culture bottles will be possible in a routine fashion with new standardized procedures. In addition to species identification, the MALDI‐TOF MS technology offers several further possibilities, like assays to detect or predict resistance phenotypes in fungi as well as subtyping approaches to detect clinically relevant subgroups. The differences between the commercial systems are discussed with respect to fungi and an overview of their performances provided. Factors influencing outcome of MALDI‐TOF‐based species identification are discussed.     

Phenoptosis in yeasts

The current view on phenoptosis and apoptosis as genetic programs aimed at eliminating potentially dangerous organisms and cells, respectively, is given. Special emphasis is placed on apoptosis (phenoptosis) in yeasts: intracellular defects and a plethora of external stimuli inducing apoptosis in yeasts; distinctive morphological and biochemical hallmarks accompanying apoptosis in yeasts; pro- and antiapoptotic factors involved in yeast apoptosis signaling; consecutive stages of apoptosis from external stimulus to the cell death; a prominent role of mitochondria and other organelles in yeast apoptosis; possible pathways for release of apoptotic factors from the intermembrane mitochondrial space into the cytosol are described. Using some concrete examples, the obvious physiological importance and expediency of altruistic death of yeast cells is shown. Poorly known aspects of yeast apoptosis and prospects for yeast apoptosis study are defined.     

Nonconventional yeasts: their genetics and biotechnological applications

To date, more than 500 species of yeasts have been described. Most of the genetic and biochemical studies have, however, been carried out with Saccharomyces cerevisiae. Although a considerable amount of knowledge has been accumulated on fundamental processes and biotechnological applications of this industrially important yeast, the large variety of other yeast genera and species may offer various advantages for experimental study as well as for product formation in biotechnology. The genetic investigation of these so-called unconventional yeasts is poorly developed and information about corresponding data is dispersed. It is the aim of this review to summarize and discuss the main results of genetic studies and biotechnological applications of unconventional yeasts and to serve as a guide for scientists who wish to enter this field or are interested in only some aspects of these yeasts.