• Title/Summary/Keyword: S cerevisiae

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Isolation and Characterization of Saccharomyces cerevisiae from nuruk for Production of Ethanol from Maltose (누룩으로부터 맥아당 이용능과 에탄올 생산성이 우수한 효모의 분리와 특성)

  • Choi, Da-Hye;Choi, Yeong-Hwan;Yeo, Soo-Hwan;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.34-39
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    • 2016
  • Wild-type yeast strains were isolated from nuruk, a type of microbial starter culture used for fermenting grains to produce alcoholic products, that was collected from different areas in Korea. Strains were identified based on the analysis of 18S rRNA sequences. Fifty strains shared the highest sequence similarity with Saccharomyces cerevisiae and were designated MBYK1-MBYK50. Among these S. cerevisiae isolates, MBYK45 produced $44.0{\pm}0.3g$ of ethanol from 200 g maltose after incubation at $30^{\circ}C$ for 48 h. Maximum ethanol production of $110.80{\pm}0.81g/l$ with productivity of $3.79{\pm}0.14g^{-1}l^{-1}h^{-1}$ was obtained at optimum culture conditions of pH (6.0), maltose (200 g/l), and temperature ($35^{\circ}C$). This study indicates that the MBYK45 strain of S. cerevisiae, isolated from nuruk, might be suitable for traditional liquor production from malts.

Analysis of Biogenic Amines Content and the Main Volatile Flavor Compounds in Black Raspberry Wine Using Traditional Yeast (토종효모를 이용한 복분자 발효주의 바이오제닉아민 함량 및 향기성분 분석)

  • Yoon, Hae-Hoon;Son, Rak-Ho;Ryu, Eun-Hye;Jung, Ji-Hye
    • Korean journal of food and cookery science
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    • v.31 no.3
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    • pp.296-303
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    • 2015
  • We investigate black raspberry (Rubus occidentalis) wine made using traditional yeast (Saccharomyces cerevisiae A8, B6, GBY2, GBY3) and S. cerevisiae Fermivin (FM), which is widely used in wine manufacturing, and analyze the biogenic amine content and the volatile flavor compounds. Black raspberries were separately inoculated with yeast up to $1{\times}10^9CFU/kg$, followed by incubation at $25^{\circ}C$ for 7 days. FM produced the highest alcohol content, however the final fermentation characteristics of the wine made using four different yeasts were similar. S. cerevisiae A8 had a large biogenic amine (BA) content, specifically tryptamine, thus we excluded this yeast from fermentation. S. cerevisiae GBY3 was selected for black raspberry wine fermentation as a result of sensory evaluation. The volatile flavor compounds of two wines (S. cerevisiae GBY3 and FM) were analyzed by gas chromatography and mass spectrometry. 37 compounds in the samples were separated, and several ester compounds were identified in greater amounts in the wine made with S. cerevisiae GBY3 than in the wine made with FM. A greater amount of the major compound, ethyl benzoate, giving the sweet and fruity flavor, was identified in wine made with S. cerevisiae GBY3 than in the wine made with FM. In conclusion, S. cerevisiae GBY3 was confirmed to produce no major BAs and a better flavored wine. These results give new leads in the production of high quality wine.

Construction of hsf1 Knockout-mutant of a Thermotolerant Yeast Strain Saccharomyces cerevisiae KNU5377 (고온내성 연료용 알코올 효모균주 Saccharomyces cerevisiae KNU5377에서 HSF1 유전자의 변이주 구축)

  • Kim Il-Sup;Yun Hae-Sun;Choi Hye-Jin;Sohn Ho-Yong;Yu Choon-Bal;Kim Jong-Guk;Jin Ing-Nyol
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.454-458
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    • 2006
  • HSF1 is the heat shock transcription factor in Saccharomyces cerevisiae. S. cerevisiae KNU5377 can ferment at high temperature such as $40^{\b{o}}C$. We have been the subjects of intense study because Hsf1p mediates gene expression not only to heat shock, but to a variety of cellular and environmental stress challenges. Basing these facts, we firstly tried to construct the hsf1 gene-deleted mutant. PCR-method for fast production of gene disruption cassette was introduced in a thermotolerant yeast S. cerevisiae KNU5377, which allowed the addition of short flanking homology region as short as 45 bp suffice to mediate homologous recombination to kanMX module. Such a cassette is composed of linking genomic DNA of target gene to the selectable marker kanMX4 that confers geneticin (G418) resistance in yeast. That module is extensively used for PCR-based gene replacement of target gene in the laboratory strains. We describe here the generation of hsf1 gene disruption construction using PCR product of selectable marker with primers that provide homology to the hsf1 gene following separation of haploid strain in wild type yeast S. cerevisiae KNU5377. Yeast deletion overview containing replace cassette module, deletion mutant construction and strain confirmation in this study used Saccharomyces Genome Deletion Project (http:://www-sequence.standard.edu/group/yeast_deletion_project). This mutant by genetic manipulation of wild type yeast KNU5377 strain will provide a good system for analyzing the research of the molecular biology underlying their physiology and metabolic process under fermentation and improvement of their fermentative properties.

Overexpression and Characterization of Bovine Pancreatic Deoxyribonuclease I in Saccharomyces cerevisiae and Pichia pastoris (Saccharomyces cerevisiae와 Pichia pastoris에서 Bovine Pancreatic Deoxyribonuclease I의 과발현과 특성)

  • Cho, Eun-Soo;Kim, Jeong-Hwan;Yoon, Ki-Hong;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.348-355
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    • 2012
  • In the present study, we investigated the overexpression and characterization of bovine pancreatic (bp)- DNase I in Saccharomyces cerevisiae and Pichia pastoris. The bp-DNase I gene was fused in frame with the GAL10 promoter, $MF{\alpha}$, and GAL7 terminator sequences, resulting in the plasmid, pGAL-$MF{\alpha}$-DNaseI (6.4 kb). Also, the bp-DNase I gene was fused in frame with the AOX1 promoter, $MF{\alpha}$, and AOX1 terminator sequences, resulting in the plasmid, pPEXI (8.8 kb). The recombinant plasmids, pGAL-$MF{\alpha}$-DNaseI and pPEXI were introduced into S. cerevisiae and P. pastoris host cells, respectively. When the transformed yeast cells were cultured at $30^{\circ}C$ for 48 h in galactose or methanol medium, bp-DNase I was overexpressed and the most of activity was found in the extracellular fraction. P. pastoris transformant activity showed 45.5 unit/mL in the culture medium at 48 h cultivation, whereas S. cerevisiae transformant revealed 37.7 unit/mL in the extracellular fraction at 48 h cultivation. The enzymatic characteristics, such as DNA cleavage and half life were investigated. Treatment of the recombinant DNase I from P. pastoris induced degradation of the calf thymus DNA within 1 minute, and this DNA degradation rate was higher than that of commercial bp-DNase I (SIGMA) and the recombinant DNase I from S. cerevisiae.

Comparison of Ethanol Fermentation by Saccharomyces cerevisiae CHY1077 and Zymomonas mobilis CHZ2501 from Starch Feedstocks (전분 기질에 대한 Saccharomyces cerevisiae CHY1077과 Zymomonas mobilis CHZ2501의 에탄올 발효 비교)

  • Choi, Giwook;Kang, Hyunwoo;Kim, Youngran;Chung, Bongwoo
    • Korean Chemical Engineering Research
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    • v.46 no.5
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    • pp.977-982
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    • 2008
  • The production of ethanol by microbial fermentation as an alternative energy source has been of interest because of increasing oil price. Saccharomyces cerevisiae and Zymomonas mobilis are two of the most widely used ethanol producers. In this study, characteristics of ethanol fermentation by Saccharomyces cerevisiae CHY1077 and Zymomonas mobilis CHZ2501 was compared. Brown rice, naked barley, and cassava were selected as representatives of the starch-based raw materials commercially available for ethanol production. The volumetric ethanol productivities by Saccharomyces cerevisiae from brown rice, naked barley and cassava were $0.68g/l{\cdot}h$, $1.03g/l{\cdot}h$ and $1.28g/l{\cdot}h$ respectively. But for the Zymomonas mobilis, $2.19g/l{\cdot}h$(brown rice), $2.60g/l{\cdot}h$(naked barley) and $3.12g/l{\cdot}h$(cassava) were obtained. Zymomonas mobilis was more efficient strain for ethanol production than S. cerevisiae.

Secretory Overexpression of Clostridium Endoglucanase A in Saccharomyces cerevisiae Using GAL10 Promoter and Exoinulinase Signal Sequeice. (Saccharomyces cerevisiae에서 GAL 10 promoter와 exoinulinase 분비 서열을 이용한 Clostridium endoglucanase A의 과발현·분비)

  • Lim, Myung-Ye;Lee, Jin-Woo;Lee, Jae-Hyung;Kim, Yeon-Hee;Seo, Jin-Ho;Nam, Soo-Wan
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1248-1254
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    • 2007
  • The secretory overexpression of Clostridium thermocellum endoglucanase A gene (celA) was examined in Saccharomyces cerevisiae using Kluyveromyces marxianus exoinulinase (INU1) signal sequence and GAL10 promoter. The two plasmids, pYEG-CT1 with its own signal sequence, and pYInu-CT1 with INU1 signal sequence were introduced to S. cerevisiae SEY2102 and S. cerevisiae 2805 host strains, respectively, and then each transformant was selected on the synthetic defined media lacking uracil. The expression level and secretion efficiency of endoglucanase A was increased by $18{\sim}22%$ and 11%, respectively, by INU1 signal sequence over celA signal sequence. By considering the high level of expression (361 unit/I), plasmid stability (89%), and secretion efficiency (70%), S. cerevisiae 2805 harboring plasmid pYInu-CT1 was selected as the opti-mal host vector system for the production of cellulose-degrading enzyme and recombinant yeast probiotic. The total expression and secretion efficiency of endoglucanase A was 418 unit/l and 73%, respectively, in the batch fermentation of S. cerevisiae 2805/pYlnu-CT1 on galactose medium. The mo-lecular weight of secreted endoglucanase A was found to be greater than 100 kDa, presumably due to the N-linked glycosylation.

Cell Surface Display of Arylsulfatase Gene from Pseudoalteromonas carageenovora in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 Pseudoalteromonas carageenovora 유래 Arylsulfatase 유전자의 표층 발현)

  • Cho, Eun-Soo;Kim, Hyun-Jin;Jung, So-A;Kim, Jeong-Hwan;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.355-360
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    • 2009
  • In this study, the arylsulfatase gene (astA, 984 bp ORF) from Pseudoalteromonas carrageenovora genome was expressed on the cell surface of S. cerevisiae by fusing with Aga2p linked to the membrane anchored protein, Aga1p. The constructed plasmid, pCTAST (7.1 kb), was introduced to S. cerevisiae EBY100 cell, and yeast transformants on YPDG plate showed the hydrolyzing activity for 4-methylumbelliferyl-sulfate and p-nitrophenyl-sulfate. When S. cerevisiae EBY100/pCTAST was grown on YPDG medium, the arylsulfatase activity of cell pellet reached about 1.2 unit/mL, whereas no extracellular arylsulfatase activity was detected. The DNA ladder in agarose prepared from agar by this recombinant arylsulfatase showed similar resolution and migration patterns with a commercial agarose. This results revealed that arylsulfatase expressed on the cell surface of S. cerevisiae could be applicable to the economic production of electrophoretic-grade agarose.

The Antioxidant and Skin-whitening Effects of Saccharomyces cerevisiae FT4-4 Isolated from Berries Grown in Sunchang (화장품 소재로서 순창 베리류 유래 Sacchromyces cerevisiae FT4-4의 항산화 활성 및 미백 효과)

  • Seo, Ji won;Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Do-Youn
    • Journal of Life Science
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    • v.31 no.2
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    • pp.175-182
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    • 2021
  • Saccharomyces lysate has the well-known function of soothing the skin in various ways: it is an anti-irritant and can treat skin care conditions, such as skin whitening and antioxidative activity. However, data on the safety for use of Saccharomyces lysate in cosmetics and skin care products are still limited. To design a new cosmetic material with antioxidant and skin-whitening effects, 80 yeast strains were isolated from berries grown in Sunchang. Among the isolates, the FT4-4 strain, which exhibited superior biological activities, was selected for further experiments. The FT4-4 strain was identified as Saccharomyces cerevisiae by 18S rRNA gene sequencing analysis. S. cerevisiae FT4-4 showed higher DPPH radical-scavenging (51.41%), superoxide dismutase (62.23%), and tyrosinase inhibition (64.75%) activities. The highest yield of biomass (3.16 g/l) and maximum growth rate of S. cerevisiae FT4-4 were observed within 16 h. Furthermore, the cytotoxicity potential of S. cerevisiae FT4-4 on B16F10 melanoma cells was measured by an MTT assay, and the results indicated that S. cerevisiae FT4-4 had a capacity to inhibit melanin up to 72.02% at an initial 10 mg/ml concentration. These results suggest that S. cerevisiae FT4-4 could be a promising candidate as a multi-functional material for application in the cosmetic industry, especially because of its antioxidant and skin-whitening effects.

Cloning and Expression of Schwanniomyces castellii Starch Gene (Schwanniomyces castellii 전분 유전자의 Cloning과 발현)

  • Park, Jong-Chun;Bai, Suk;Chun, Soon-Bai
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.653-659
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    • 1990
  • The gene encoding glucoamylase from Schwanniomyces cagtellii CBS 2863 was cloned and expressed in Saccharomyces cerevisiae. Southern blot analysis confirmed that this glucoamylase gene was derived from the genomic DNA of Schwanniomyces ccastellii and that no DNA fragments corresponding to 5.1 or 1.3 kb of Sch. casteltii DNA were detected in S. cereuisiae. The glucoamylase activity from S. cerevisiae transformant was approximately 2,000 times less than that of donor yeast. No expression was found in E. coti. The secreted glucoamylase from S. cerevisiae transformant was indistinguishable from that of Sch. eastellii on the basis of molecular weight and enzyme properties.

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Size-based Separation of Yeast Cell by Surface Acoustic Wave-induced Acoustic Radiation Force (음향방사력을 이용한 효모세포의 크기별 분리)

  • Raihan Hadi Julio;Muhammad Soban Khan;Mushtaq Ali;Ghulam Destgeer;Jinsoo Park
    • Journal of the Korean Society of Visualization
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    • v.21 no.3
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    • pp.93-100
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    • 2023
  • The yeast Saccharomyces cerevisiae (S. cerevisiae) is considered an ideal eukaryotic model and has long been recognized for its pivotal role in numerous industrial production processes. Depending on the cell cycle phases, microenvironment, and species, S. cerevisiae varies in shape and has different sizes of each shape such as singlets, doublets, and clusters. Obtaining high-purity populations of uniformly shaped S. cerevisiae cells is crucial in fundamental biological research and industrial operations. In this study, we propose an acoustofluidic method for separating S. cerevisiae cells based on their size using surface acoustic wave (SAW)-induced acoustic radiation force (ARF). The SAW-induced ARF increased with cell diameter, which enabled a successful size-based separation of S. cerevisiae cells using an acoustofluidics device. We anticipate that the proposed acoustofluidics approach for yeast cell separation will provide new opportunities in industrial applications.