• Title/Summary/Keyword: non-Saccharomyces

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The Antifungal Test: An Efficient Screening Tool for the Discovery of Microbial Metabolites with Respiratory Inhibitory Activity

  • Han, Jae Woo;Kim, Bomin;Oh, Mira;Choi, Jaehyuk;Choi, Gyung Ja;Kim, Hun
    • Mycobiology
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    • v.48 no.4
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    • pp.326-329
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    • 2020
  • Valuable natural compounds produced by a variety of microorganisms can be used as lead molecules for development of new agrochemicals. Furthermore, high-throughput in vitro screening systems with specific modes of action can increase the probability of discovery of new fungicides. In the current study, a rapid assay tested with various microbes was developed to determine the degree of respiratory inhibition of Saccharomyces cerevisiae in two different liquid media, YG (containing a fermentable carbon source) and NFYG (containing a non-fermentable carbon source). Based on this system, we screened 100 fungal isolates that were classified into basidiomycetes, to find microbial secondary metabolites that act as respiratory inhibitors. Consequently, of the 100 fungal species tested, the culture broth of an IUM04881 isolate inhibited growth of S. cerevisiae in NFYG medium, but not in YG medium. The result is comparable to that from treatment with kresoxim-methyl used as a control, suggesting that the culture broth of IUM04881 isolate might contain active compounds showing the inhibition activity for respiratory chain. Based on the assay developed in this study and spectroscopic analysis, we isolated and identified an antifungal compound (-)-oudemansin A from culture broth of IUM04881 that is identified as Oudemansiella venosolamellata. This is the first report that (-)-oudemansin A is identified from O. venosolamellata in Korea. Taken together, the development of this assay will accelerate efforts to find and identify natural respiratory inhibitors from various microbes.

Evaluation of Yeast Diversity During Wine Fermentations with Direct Inoculation and pied de cuve Method at an Industrial Scale

  • Li, Erhu;Liu, Chuanhe;Liu, Yanlin
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.960-966
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    • 2012
  • The diversity and composition of yeast populations may greatly impact wine quality. This study investigated the yeast microbiota in two different types of wine fermentations: direct inoculation of a commercial starter versus pied de cuve method at an industrial scale. The pied de cuve fermentation entailed growth of the commercial inoculum used in the direct inoculation fermentation for further inoculation of additional fermentations. Yeast isolates were collected from different stages of wine fermentation and identified to the species level using Wallersterin Laboratory nutrient (WLN) agar followed by analysis of the 26S rDNA D1/D2 domain. Genetic characteristics of the Saccharomyces cerevisiae strains were assessed by a rapid PCR-based method, relying on the amplification of interdelta sequences. A total of 412 yeast colonies were obtained from all fermentations and eight different WL morphotypes were observed. Non-Saccharomyces yeast mainly appeared in the grape must and at the early stages of wine fermentation. S. cerevisiae was the dominant yeast species using both fermentation techniques. Seven distinguishing interdelta sequence patterns were found among S. cerevisiae strains, and the inoculated commercial starter, AWRI 796, dominated all stages in both direct inoculation and pied de cuve fermentations. This study revealed that S. cerevisiae was the dominant species and an inoculated starter could dominate fermentations with the pied de cuve method under controlled conditions.

Complete Sequence of a Gene Encoding KAR3-Related Kinesin-like Protein in Candida albicans

  • Kim Min-Kyoung;Lee Young Mi;Kim Wankee;Choi Wonja
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.406-410
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    • 2005
  • In contrast to Saccharomyces cerevisiae, little is known about the kinesin-like protein (KLP) in Candida albicans. The motor domain of kinesin, or KLP, contains a subregion, which is well conserved from yeast to humans. A similarity search, with the murine ubiquitous kinesin heavy chain region as a query, revealed 6 contigs that contain putative KLPs in the genome of C. albicans. Of these, the length of an open reading (ORF) of 375 amino acids, temporarily designated CaKAR3, was noticeably short compared with the closely related S. cerevisiae KAR3 (ScKAR3) of 729 amino acids. This finding prompted us to isolate a ${\lambda}$ genomic clone containing the complete CaKAR3 ORF, and here the complete sequence of CaKAR3 is reported. CaKAR3 is a C-terminus motor protein, of 687 amino acids, encoded by a non-disrupting gene. When compared with ScKAR3, the amino terminal region of 112 amino acids was unique, with the middle part of the 306 amino acids exhibiting $25\%$ identity and $44\%$ similarity, while the remaining C-terminal motor domain exhibited $64\%$ identity and $78\%$ similarity, and have been submitted to GeneBank under the accession number AY182242.

Effect of fermented spent instant coffee grounds on milk productivity and blood profiles of lactating dairy cows

  • Choi, Yongjun;Rim, Jongsu;Lee, Honggu;Kwon, Hyunchul;Na, Youngjun;Lee, Sangrak
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.1007-1014
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    • 2019
  • Objective: This study was conducted to evaluate the fermentation characteristics under low mesophilic temperature of spent instant coffee ground (SICG) and to estimate the effect of fermented SICG (FSICG) as alternative feed ingredient on milk productivity of dairy cows. Methods: In the fermentation trial, fermentation of SICG was performed to investigate changes in characteristics using the microbial mixture (Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis = 1:1:1) for 21 days at $20^{\circ}C$ under anaerobic conditions. Molasses was added at 5% of dry mass. In the animal trial, eighteen Holstein Friesian cows were used to evaluate the nutritive value of the FSICG which was fermented for 14 days under the same condition as the fermentation trial. Results: In the fermentation trial, the dry matter (DM) and organic matter content linearly decreased with fermentation time (p<0.001 and p = 0.008, respectively). The acid detergent insoluble nitrogen content linearly decreased with fermentation time (p = 0.037). The microorganism counts linearly increased for Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis across fermentation time (p<0.001). In the animal trial, the DM intake of the control and FSICG treatment were not significantly different, as were milk yield, 4% fat corrected milk, fat-protein corrected milk, and feed to milk conversion content. Fat, protein, lactose, non-fat solids, milk urea nitrogen, and somatic cell counts were also not significantly different in milk composition between treatments. Conclusion: FSICG should be considered a sufficient substitute for cottonseed as a feed component, and 5% DM of a dietary FSICG level was appropriate for dairy cow diets.

Microbiological Characteristics of Wild Yeast Strain Pichia anomala Y197-13 for Brewing Makgeolli

  • Kim, Hye Ryun;Kim, Jae-Ho;Bai, Dong-Hoon;Ahn, Byung Hak
    • Mycobiology
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    • v.41 no.3
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    • pp.139-144
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    • 2013
  • Makgeolli is a traditional cloudy-white Korean rice wine with an alcohol content of 6~7%. The present study investigated the morphological characteristics, carbon-utilizing ability, fatty acid composition, alcohol resistance, glucose tolerance, and flocculence of Saccharomyces cerevisiae Y98-5 and Pichia anomala Y197-13, non-S. cerevisiae isolated from Nuruk, which is used in brewing Makgeolli. Similar morphological characteristics were observed for both isolated wild yeast strains; and the carbon source assimilation of Y197-13 differed from that of other P. anomala strains. Strain Y197-13 was negative for D-trehalose, mannitol, arbutin, I-erythritol, and succinic acid. The major cellular fatty acids of strain Y197-13 included C18:2n6c (33.94%), C18:1n9c (26.97%) and C16:0 (20.57%). Strain Y197-13 was Crabtree-negative, with 60% cell viability at 12% (v/v) ethanol. The flocculation level of strain Y197-13 was 8.38%, resulting in its classification as a non-flocculent yeast.

Molecular Genetic Identification of Yeast Strains Isolated from Egyptian Soils for Solubilization of Inorganic Phosphates and Growth Promotion of Corn Plants

  • Hesham, Abd El-Latif;Mohamed, Hashem M.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.55-61
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    • 2011
  • Forty yeast strains isolated from soils taken from different locations in Egypt were tested for their P-solubilizing activities on the basis of analyzing the clear zone around colonies growing on a tricalcium phosphate medium after incubation for 5 days at $25^{\circ}C$, denoted as the solubilization index (SI). Nine isolates that exhibited P-solubilization potential with an SI ranging from 1.19 to 2.76 were genetically characterized as five yeasts belonging to the genus Saccharomyces cerevisiae and four non-Saccharomyces, based on a PCR analysis of the ITS1-26S region amplied by SC1/SC2 species-specific primers. The highest P-solubilization efficiency was demonstrated by isolate PSY- 4, which was identified as Saccharomyces cerevisiae by a sequence analysis of the variable D1/D2 domain of the 26S rDNA. The effects of single and mixed inoculations with yeast PSY-4 and Bacillus polymyxa on the P-uptake and growth of corn were tested in a greenhouse experiment using different levels of a phosphorus chemical fertilizer (50, 100, and 200 kg/ha super phosphate 15.5% $P_2O_5$). The results showed that inoculating the corn with yeast PSY-4 or B. polymyxa caused significant increases in the shoot and root dry weights and P-uptake in the shoots and roots. The P-fertilization level also had a significant influence on the shoot and root dry weights and P-uptake in the shoots and roots when increasing the P-level from 50 up to 200 kg/ha. Dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 200 kg/ha gave higher values for the shoot and root dry weights and P-uptake in the shoots and roots, yet these increases were nonsignificant when compared with dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha. The best increases were obtained from dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha, which induced the following percentage increases in the shoot and root dry weights, and P-uptake in the shoots and roots; 16.22%, 46.92%, 10.09%, and 31.07%, respectively, when compared with the uninoculated control (fertilized with 100 kg/ha).

Selection of Constitutive Promoter for Exoinulinase Production in Fed-Batch Culture of Recombinant Yeast (재조합 효모의 유가배양에서 Exoinulinase생산을 위한 Promoter의 선별)

  • 김이경;고지현;김연희;김성구;남수완
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.206-211
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    • 2001
  • In order to overexpress constitutively the Kluyveromyces marxianus exoinulinase gene (INUI) in Saccharomyces cerevisiae, four episomal expression systems employing GAPDH, ADHI, PGK and ENOI promoters were constructed as p YIGP aADHI -INU, pPGK-INU, and pENOI- INU plasmids respectively, When S cereviais transformants harboring each plasmid were batchwisely cultivated in the fermentor containing 5% glucose medium no significant differences in the cell growth are observed How- ever the experession level of exoinulinase and plasmid stability showed a strong dependency on the promoter employed. The expression levels of exoinulinase were about 1.70 unit/ml for GAPDH promoter 1.67 unit/ml for PGK promoter, 1.29 unit /ml for ADH1 promoter, and 0.80 unit/ml for ENOl promoter. The plasmid stabilites were maintaines above 80% in all experession systems. except the GAPDH promoter system of 55%, Based on the plas- mid stability and expression level of exoinulinase the ADHl and PGK promoter system were selected for the fed - batch culture to overproduce exoinulinase By the intermittent feeding of yeast extract and glucose, both promoter systems gave the cell concentration of about 30 g-dry cell weight/1 byt the maximal exoinulinase activity of 3.70 unit/ml and plasmid stability of 96% in the ADH1 promoter were higher than those (2.70 unit/ml, 80%) of PGK sys- tem Taking into account the plasmid stability and extended culture time the ADH1 promoter systems would be the most feasible expression systems for the constitutive overproduction of exoinulinase through high cell-density fed- batch cultures using non-selective rich medium.

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Protective Effects of Zizyphus jujuba and Fermented Zizyphus jujuba from Free Radicals and Hair Loss (대추 및 발효대추의 라디칼 소거능 및 모발 성장 촉진 효과)

  • Jung, Ji Eun;Cho, Eun Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1174-1180
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    • 2014
  • This study investigated the antioxidative and hair growth-promoting activities of Zizyphus jujuba (Zj) and fermented Zj with Aspergillus oryzae, Bacillus subtilis, Bifidobacterium breve, Lactobacillus acidophilus, and Saccharomyces cerevisiae (Sac. cerevisiae). Among Zj and fermented Zjs, Sac. cerevisiae-fermented Zj (Zj-Y) exerted stronger scavenging activity against 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radicals than others. In addition, total polyphenol content of Zj-Y was higher than that of non-fermented Zj and other fermented Zj. This result indicates that fermentation of Zj by Sac. cerevisiae elevated antioxidative activity. Furthermore, using an alopecia model in C57B/6N mice, the hair growth activities of Zj and Zj-Y were investigated. The test samples, EtOH, minoxidil (MXD), Zj, and Zj-Y, were topically treated with 0.2 mL/day for 4 weeks. The experiments involved macroscopic observation and measurement of hair length methods. The results show that regrowth speed of hair was in decreasing order of MXD> Zj-Y> Zj> EtOH. The topical application of MXD and Zj-Y in mice promoted hair regrowth and prevented hair loss compared to the control group. The present study indicates that Zj-Y is a promising treatment for alopecia.

Effects of low temperature-adapted Saccharomyces cerevisiae Y297 strain and fermentation temperature on the quality characteristics of Yakju (저온 적응성 효모와 발효온도에 따른 약주의 품질특성 변화)

  • Seo, Dong-Jun;Yeo, Soo-Hwan;Mun, Ji-Young;Baek, Seong Yeol
    • Food Science and Preservation
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    • v.23 no.5
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    • pp.666-672
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    • 2016
  • The objective of this study was to evaluate the effects of low temperature-adapted Saccharomyces cerevisiae Y297 and fermentation temperatures on the quality of Yakju brewed. Physicochemical properties of Yakju brewed were compared pH, total acidity, ethanol, free amino acid, organic acid contents, and volatile flavor compounds in S. cerevisiae Y297 with control treatment. Cooked non-glutinous rice and saccharogenic amylase in koji were mixed with ethanol-producing yeasts and then fermented at $15^{\circ}C$, $20^{\circ}C$, and $25^{\circ}C$ for 20 days. Yakju brewed using the Y297 treatment showed the highest ethanol yield (17.9%) at $20^{\circ}C$. Expression of heat shock protein (HSP) 104 was evaluated by immunoblotting as an indication of adaptation to low temperatures ($15^{\circ}C$); levels of the HSP104 protein were higher in the Y297 treatment than in the control. Organic acid analysis showed that the lactic acid content of Yakju brewed using the control was the highest at $25^{\circ}C$. Finally, free amino acid analysis showed that the Y297 treatment had a higher proportion of essential amino acids than the control. Overall, these results indicate that S. cerevisiae Y297 could be used as a suitable yeast for Yakju brewed under low temperature ($15^{\circ}C$) condition.

Characterization of acetic acid fermentation of detoxified Rhus verniciflua vinegar produced by various yeast strains (효모 종류에 따른 무독화 옻식초의 초산발효 특성)

  • Kim, Ji-Seon;Yeo, Soo-Hwan;Mun, Ji-Young;Baek, Seong Yeol
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.1042-1049
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    • 2016
  • This study was conducted to investigate and compare the characteristics of acetic acid fermentation in detoxified Rhus verniciflua vinegar (DRV) produced by different yeast strains. The DRVs were prepared by static acetic acid fermentation using six different yeast strains (Saccharomyces cerevisiae Fermivin, Pichia kudriavzerii N77-4, Hanseniaspora pountiae HP1-2, Candida tropicalis Y447, Wickerhamomyces anomalus N43-8, and Pichia kluyveri Frootzen). Alcohol content of the S. cerevisiae Fermivin fermented DRV was highest 16.07%. Among the yeast strain DRVs, there were significant differences in alcohol content, but all alcohol levels were 11%. Moreover, there were differences in pH and titratable acidity of the DRVs. The organic acid content of the DRVs ranged from 35.88 to 55.49 mg/mL and there were significant differences among the yeast strain DRVs. Essential free amino acids, particularly glutamic acid, alanine, leucine and valine, were detected in each of the 6 DRVs. Electronic nose analysis revealed that three different volatile chemical patterns were present in the 6 DRVs. The results indicate that yeast strains with different characteristics can produce vinegars with different characteristics.