We have previously isolated epsilon-COP, the alpha-COP interactor in COPI of Aspergillus nidulans, by yeast two-hybrid screening. To understand the function of epsilon-COP, the aneA+ gene for epsilon-COP/AneA was deleted by homologous recombination using a gene-specific disruption cassette. Deletion of the epsilon-COP gene showed no detectable changes in vegetative growth or asexual development, but resulted in decrease in the production of the fruiting body, cleistothecium, under conditions favorable for sexual development. Unlike in the budding yeast Saccharomyces cerevisiae, in A. nidulans, over-expression of epsilon-COP did not rescue the thermo-sensitive growth defect of the alpha-COP mutant at 42degrees C. Together, these data show that epsilon-COP is not essential for viability, but it plays a role in fruiting body formation in A. nidulans.
Aspergillus nidulans* ; Coatomer Protein* ; Fruit ; Homologous Recombination ; Mass Screening ; Saccharomyces cerevisiae ; Saccharomycetales ; Sexual Development ; Yeasts

Two-hundred two yeast strains were isolated from rhizosphere (87 strains) and nonrhizosphere (115 strains) areas of potato, maize, vegetable marrow, and cabbage plants. On the basis of 26 morphological and physiological properties, the isolated yeast strains were assigned to 9 genera and 15 species. Trichosporon beigelii, Kluyveromyces marxianus and Torulaspora delbrueckii were the dominant species. Cryptococcus humicolus and Candida tropicalis were represented by considerable numbers of strains. Of low occurrence were Saccharomyces cerevisiae and Candida blankii. Other yeast species were represented by single or two strains. Total counts of yeast cells per gram dry soil ranged from 1.1x10(3) to 6.6x10(3) in soil samples of rhizosphere areas and from 6.5x10(2) to 5.6x10(3) in soil samples of nonrhizosphere areas. Types of the tested plants affected not only the total counts of yeast cells but also spectra of yeast species. Relationships of age of potato plant, moisture contents of soil samples, and its pH values and total counts of yeast cells were discussed.
Bone Marrow ; Brassica ; Candida ; Candida tropicalis ; Cryptococcus ; Egypt* ; Hydrogen-Ion Concentration ; Kluyveromyces ; Plants ; Rhizosphere ; Saccharomyces cerevisiae ; Soil* ; Solanum tuberosum ; Torulaspora ; Trichosporon ; Vegetables ; Yeasts* ; Zea mays

Msi1-like (MSIL) proteins, which are eukaryote-specific and contain a series of WD40 repeats, have pleiotropic roles in chromatin assembly, DNA damage repair, and regulation of nutrient/stress-sensing signaling pathways. In the fungal kingdom, the functions of MSIL proteins have been studied most intensively in the budding yeast model Saccharomyces cerevisiae, an ascomycete. Yet their functions are largely unknown in other fungi. Recently, an MSIL protein, Msl1, was discovered and functionally characterized in the pathogenic yeast Cryptococcus neoformans, a basidiomycete. Interestingly, MSIL proteins appear to have redundant and unique roles in both fungi, suggesting that MSIL proteins may have evolutionarily divergent roles in different parts of the fungal kingdom. In this review, we will describe the current findings regarding the role of MSIL proteins in fungi and discuss future directions for research on this topic.
Ascomycota ; Basidiomycota ; Chromatin Assembly and Disassembly ; Cryptococcus neoformans ; DNA Damage ; Fungi ; Histones ; Proteins ; Retinoblastoma ; Saccharomyces cerevisiae ; Saccharomycetales ; Yeasts

Kluyveromyces fragilis KCTC 7260 and Saccharomyces cerevisiae KCTC 7904, which both grew well in pear marc extract, were selected and their growth profiles and physiological functionalities were determined. Both of the selected yeasts established maximal growth by 20 hr of cultivation at 30degrees C in pear marc extract. The cell-free extracts showed high antihypertensive angiotensin I-converting enzyme inhibitory activity of 68.9% and 52.1%, respectively. The extracts also displayed 9.2 U/mL and 12.0 U/mL of protease activity, respectively.
Kluyveromyces ; Peptidyl-Dipeptidase A ; Pyrus ; Saccharomyces cerevisiae ; Yeasts

The target of rapamycin (TOR) signaling pathway conserved from yeast to human plays critical roles in regulation of eukaryotic cell growth. It has been shown that TOR pathway is involved in several cellular processes, including ribosome biogenesis, nutrient response, autophagy and aging. However, due to the functional diversity of TOR pathway, we do not know yet some key effectors of the pathway. To find unknown effectors of TOR signaling pathway, we took advantage of a green fluorescent protein (GFP)-tagged collection of budding yeast Saccharomyces cerevisiae . We analyzed protein abundance changes by measuring the GFP fluorescence intensity of 4156 GFP-tagged yeast strains under inhibition of TOR pathway. Our proteomic analysis argues that 83 proteins are decreased whereas 32 proteins are increased by treatment of rapamycin, a specific inhibitor of TOR complex 1 (TORC1). We found that, among the 115 proteins that show significant changes in protein abundance under rapamycin treatment, 37 proteins also show expression changes in the mRNA levels by more than 2-fold under the same condition. We suggest that the 115 proteins indentified in this study may be directly or indirectly involved in TOR signaling and can serve as candidates for further investigation of the effectors of TOR pathway.
Aging ; Autophagy ; Eukaryotic Cells ; Fluorescence ; Humans ; Imidazoles ; Multiprotein Complexes ; Nitro Compounds ; Proteins ; Proteome ; Ribosomes ; RNA, Messenger ; Saccharomyces cerevisiae ; Saccharomycetales ; Sirolimus ; TOR Serine-Threonine Kinases ; Yeasts ; Organelle Biogenesis

Wild yeasts on the surface of various fruits including grapes were surveyed to obtain yeast strains suitable for fermenting a novel wine with higher alcohol content and supplemented with rice starch. We considered selected characteristics, such as tolerance to alcohol and osmotic pressure, capability of utilizing maltose, and starch hydrolysis. Among 637 putative yeast isolates, 115 strains exhibiting better growth in yeast-peptone-dextrose broth containing 30% dextrose, 7% alcohol, or 2% maltose were selected, as well as five alpha-amylase producers. Nucleotide sequence analysis of the 26S rDNA gene classified the strains into 13 species belonging to five genera; Pichia anomala was the most prevalent (41.7%), followed by Wickerhamomyces anomalus (19.2%), P. guilliermondii (15%), Candida spp. (5.8%), Kodamaea ohmeri (2.5%), and Metschnikowia spp. (2.5%). All of the alpha-amylase producers were Aureobasidium pullulans. Only one isolate (NK28) was identified as Saccharomyces cerevisiae. NK28 had all of the desired properties for the purpose of this study, except alpha-amylase production, and fermented alcohol better than commercial wine yeasts.
alpha-Amylases ; Base Sequence ; Candida ; DNA, Ribosomal ; Fermentation ; Fruit ; Glucose ; Hydrolysis ; Maltose ; Mass Screening ; Metschnikowia ; Osmotic Pressure ; Pichia ; Saccharomyces cerevisiae ; Starch ; Vitis ; Wine ; Yeasts

No abstract available.
Candidemia* ; Saccharomycetales*

Pichia pastoris is one of the most widely used recombinant protein expression systems. In this study, a novel method for rapid screening of P. pastoris strains capable of efficiently expressing recombinant proteins was developed. Firstly, the ability to express recombinant proteins of the modified strain GS115-E in which a functional Sec63-EGFP (Enhanced green fluorescent protein) fusion protein replaced the endogenous endoplasmic reticulum transmembrane protein Sec63 was tested. Next, the plasmids carrying different copy numbers of phytase (phy) gene or xylanase (xyn) gene were transformed into GS115-E to obtain recombinant strains with different expression levels of phytase or xylanase, and the expression levels of EGFP and recombinant proteins in different strains were tested. Finally, a flow cytometer sorter was used to separate a mixture of cells with different phytase expression levels into sub-populations according to green fluorescence intensity. A good linear correlation was found between the fluorescence intensities of EGFP and the expression levels of the recombinant proteins in the recombinant strains (0.8<|R|<1). By using the flow cytometer, high-yielding P. pastoris cells were efficiently screened from a mixture of cells. The expression level of phytase of the selected high-fluorescence strains was 4.09 times higher than that of the low-fluorescence strains after 120 h of methanol induction. By detecting the EGFP fluorescence intensity instead of detecting the expression level and activity of the recombinant proteins in the recombinant strains, the method developed by the present study possesses the greatly improved performance of convenience and versatility in screening high-yielding P. pastoris strains. Combining the method with high-throughput screening instruments and technologies, such as flow cytometer and droplet microfluidics, the speed and throughput of this method will be further increased. This method will provide a simple and rapid approach for screening and obtaining P. pastoris with high abilities to express recombinant proteins.
6-Phytase/genetics* ; Pichia/genetics* ; Plasmids ; Recombinant Proteins/genetics* ; Saccharomycetales

Nowadays, engineered Komagataella phaffii plays an important role in the biosynthesis of small molecule metabolites and protein products, showing great potential and value in industrial productions. With the development and application of new editing tools such as CRISPR/Cas9, it has become possible to engineer K. phaffii into a cell factory with high polygenic efficiency. Here, the genetic manipulation techniques and objectives for engineering K. phaffii are first summarized. Secondly, the applications of engineered K. phaffii as a cell factory are introduced. Meanwhile, the advantages as well as disadvantages of using engineered K. phaffii as a cell factory are discussed and future engineering directions are prospected. This review aims to provide a reference for further engineering K. phaffii cell factory, which is supposed to facilitate its application in bioindustry.
Saccharomycetales/genetics* ; Genetic Techniques

Aims: Oil palm trunk (OPT) can be a potential biomass from replanting activities for biomass-to-liquid (BTL) particularly in bioethanol production. The OPT contains higher carbohydrates compared to other oil palm biomass, thus has better advantages as feedstock for biofuel. To realise this, the feasibility of using oil palm trunk (OPT) sap as a substrate for bioethanol fermentation was explored via optimising the various culture conditions (pH, temperature, inoculum size, nitrogen source, dilution effect and growth medium) using Saccharomyces cerevisiae. Methodology and results: A total of six parameters were tested for optimising bioethanol production i.e. pH, temperature, inoculum size, nitrogen source, dilution effect and types of medium. Results showed that the optimum conditions for OPT sap in bioethanol production were at pH 4.0, temperature of 30 °C, inoculum size of 10 % (v/v), without requirement of nitrogen supplementation and substrate dilution. A fermentation period of 24 h was best for bioethanol production and resulted in bioethanol production, formation rate and yield of 47.5 g/L, 1.98 g/h and 0.50 g/g, respectively. Conclusion, significance and impact study: The study has clearly demonstrated that high efficient bioethanol production from OPT sap is possible but it is susceptible to various fermentation influencing parameters. This study could establish an effective and sustainable utilisation of waste OPT especially its sap as a lignocellulosic biomass supplement from the oil palm industry for second generation biofuel production.
Saccharomyces cerevisiae ; Fermentation