• Journal of Microbiology and Biotechnology
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• 제26권4호
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• pp.700-709
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• 2016

The mitogen-activated protein kinase HOG1 (high-osmolarity glycerol response pathway) plays a crucial role in the response of yeast to hyperosmotic shock. Trichosporonoides oedocephalis produces large amounts of polyols (e.g., erythritol and glycerol) in a culture medium. However, the effects of HOG1 gene knockout and environmental stress on the production of these polyols have not yet been studied. In this study, a To-HOG1 null mutation was constructed in T. oedocephalis using the loxP-Kan-loxP/Cre system as replacement of the targeted genes, and the resultant mutants showed much smaller colonies than the wild-type controls. Interestingly, compared with the wild-type strains, the results of shake-flask culture showed that To-HOG1 null mutation increased erythritol production by 1.44-fold while decreasing glycerol production by 71.23%. In addition, this study investigated the effects of citric acid stress on the T. oedocephalis HOG1 null mutants and the wild-type strain. When the supplementation of citric acid in the fermentation medium was controlled at 0.3% (w/v), the concentration of erythritol produced from the wild-type and To-HOG1 knockout mutant strains improved by 18.21% and 21.65%, respectively.

• BMB Reports
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• 제31권6호
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• pp.572-577
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• 1998

A soluble protein from Saccharomyces cerevisiae provides protection against a thiol-containing oxidation system but not against an oxidation system without thiol. This 25-kDa protein acts as a peroxidase but requires the NADPH-dependent thioredoxin system or a thiol-containing intermediate, and was thus named thioredoxin peroxidase. The protective role of thioredoxin peroxidase against ionizing radiation, which generates reactive oxygen species harmful tocellular function, was investigated in wild-type and mutant yeast strains in which the tsa gene encoding thioredoxin peroxidase was disrupted by homologous recombination. Upon exposure to ionizing radiation, there was a distinct difference between these two strains in regard to viability and the level of protein carbonyl content, which is the indicative marker of oxidative damage to protein. Activities of other antioxidant enzymes, such as catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione reductase were increased at 200-600 Gy of irradiation in wild-type cells. However, the activities of antioxidant enzymes were not significantly changed by ionizing radiation in thioredoxin peroxidase-deficient mutant cells. These results suggest that thioredoxin peroxidase acts as an antioxidant enzyme in cellular defense against ionizing radiation through the removal of reactive oxygen species as well as in the protection of antioxidant enzymes.

• Mycobiology
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• 제49권6호
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• pp.582-588
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• 2021

The interaction of mating pheromone and pheromone receptor from the B mating-type locus is the first step in the activation of the mushroom mating signal transduction pathway. The B mating-type locus of Lentinula edodes is composed of Bα and Bβ subloci, each of which contains genes for mating pheromone and pheromone receptor. Allelic variations in both subloci generate multiple B mating-types through which L. edodes maintains genetic diversity. In addition to the B mating-type locus, our genomic sequence analysis revealed the presence of a novel chromosomal locus 43.3 kb away from the B mating-type locus, containing genes for a pair of mating pheromones (PHBN1 and PHBN2) and a pheromone receptor (RCBN). The new locus (Bα-N) was homologous to the Bα sublocus, but unlike the multiallelic Bα sublocus, it was highly conserved across the wild and cultivated strains. The interactions of RcbN with various mating pheromones from the B and Bα-N mating-type loci were investigated using yeast model that replaced endogenous yeast mating pheromone receptor STE2 with RCBN. The yeast mating signal transduction pathway was only activated in the presence of PHBN1 or PHBN2 in the RcbN producing yeast, indicating that RcbN interacts with self-pheromones (PHBN1 and PHBN2), not with pheromones from the B mating-type locus. The biological function of the Bα-N locus was suggested to control the expression of A mating-type genes, as evidenced by the increased expression of two A-genes HD1 and HD2 upon the treatment of synthetic PHBN1 and PHBN2 peptides to the monokaryotic strain of L. edodes.

• KSBB Journal
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• 제17권3호
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• pp.290-294
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• 2002

경제적으로 에리스리톨을 대량 생산하기 위해 Moniliella suaveolens var. nigra의 wild type을 mutation시켜 변이주를 선별하였다. 선별된 변이주는 400 g/L 포도당배지 플라스크 배양에서 에리스리톨 172 g/L, 글리세롤 20 g/L를 생산하였는데 wild-type의 배양결과와 비교하면 에리스리롤의 생산성은 비슷하고 부산물인 글리세롤의 생성은 50% 적다. 변이주는 wild type 배양시 발생하는 다량의 foam을 적게 발생시킴이 관찰되었다. 배양기 5 L 배양에서 변이주는 에리스리톨의 생산과 부산물 글리세롤의 생성이 wild-type의 것들과 비슷하였다. 이의 원인은 아직 규명되지 않았으나 배양 중 부적절한 산소전달이나 배양 중 발생하는 거품에 세포가 응집하였기 때문인 것으로 추측된다. 균주의 대사를 조사한 결과, 글리세롤은 에리스리톨로 변환되나 에리스리톨은 글리세롤로 변환되지 않음이 관찰되었다. 새로운 고생산성 Moniliella 변이주는 분리.정제 비용이 절감된 순도 놀은 에리스리톨 생성을 가능 하게 할 것이다.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.761-767
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• 2022

EHT1 and EEB1 are the key Saccharomyces cerevisiae genes involved in the synthesis of ethyl esters during wine fermentation. We constructed single (Δeht1, Δeeb1) and double (Δeht1Δeeb1) heterogenous mutant strains of the industrial diploid wine yeast EC1118 by disrupting one allele of EHT1 and/or EEB1. In addition, the aromatic profile of wine produced during fermentation of simulated grape juice by these mutant strains was also analyzed. The expression levels of EHT1 and/or EEB1 in the relevant mutants were less than 50% of the wild-type strain when grown in YPD medium and simulated grape juice medium. Compared to the wild-type strain, all mutants produced lower amounts of ethyl esters in the fermented grape juice and also resulted in distinct ethyl ester profiles. ATF2, a gene involved in acetate ester synthesis, was expressed at higher levels in the EEB1 downregulation mutants compared to the wild-type and Δeht1 strains during fermentation, which was consistent with the content of acetate esters. In addition, the production of higher alcohols was also markedly affected by the decrease in EEB1 levels. Compared to EHT1, EEB1 downregulation had a greater impact on the production of acetate esters and higher alcohols, suggesting that controlling EEB1 expression could be an effective means to regulate the content of these aromatic metabolites in wine. Taken together, the synthesis of ethyl esters can be decreased by deleting one allele of EHT1 and EEB1 in the diploid EC1118 strain, which may modify the ester profile of wine more subtly compared to the complete deletion of target genes.

• Journal of Microbiology
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• 제41권2호
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• pp.121-128
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• 2003

The identification of genes involved in true hypha formation is important in the study of mechanisms underlying the morphogenetic switch in yeast. We isolated a gene responsible for the morphogenetic switch in Yarrowia lipolytica, which forms true hyphae in response to serum or N-acetylglucosamine. The isolated gene, encoding 847 amino acids, had sequence identities of 27% and 25% with the Bud6 (Aip3) proteins of Saccharomyces cerevisiae and Schizosaccharomyces pombe, respectively. Disruption of this gene, designated YIBUD6, in haploid and diploid strains significantly reduced the ability of Y. lipolytica to switch from the yeast form to the hyphal form in hypha-inducing media. It was also found that YIBud6$\Delta$ mutants were rounder than the wild type when grown in the yeast form. These results indicate that the YIBud6 protein is necessary for hyphal growth and cell polarity in both haploid and diploid Y. lipolytica cells.

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.668-671
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• 1999

The gene encoding Schwanniomyces occidentalis $\alpha-amylase$(AMY) was introduced into Saccharomyces cerevisiae var. diastaticus which secreted only glucoamylase, by using a linearized yeast integrating vector to develop stable strains with a capability of secreting $\alpha-amylase$and glucoamylase simultaneously. A dominant selectable marker, the geneticin(G418) resistance gene (Gt^r$), was cloned into a vector to screen wild-type diploid transformants harboring the AMY gene. The amylolytic activities of transformants were about 3-7 times higher than those of the recipient strains. When grown in nonselective media, the transformants with the linearized integrating vector containing the AMY gene exhibited almost all of the mitotic stability after 100 generations.

• 생명과학회지
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• 제16권3호
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• pp.454-458
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• 2006

출아효모인 Sacharomyces cerevisiae S288C균주를 이용한 효모의 게놈이 완성된 후 S. cerevisiae는 다양한 연구 모델로 이용되어져 왔다. 현재까지 효모를 이용한 기능 유전체학 측면에서의 연구는 laboratory strainin인 S288C 균주 또는 그 유래의 균주들이다. 그러나 자연에서 분리된 효모 또는 산업적으로 이용되어지고 있는 S. cerevisiae의 유전학 측면에서의 연구는 낮은 포자형성률 및 형질전환률, 그리고 S288C 균주와의 게놈상의 상이성 때문에 거의 이루어지지 않고 있다. 여기서 우리 연구진은 자연에서 분리된 Saccharomyces cerevisiae KNU5377 균주를 이용하여 random spore analysis를 통해 MATa 및 $MAT{\alpha}$ 타입의 각각의 haploid cell을 분리 후 이미 보고된 KanMX module를 가지고 round PCR기법에 의한 short flanking homology 기법을 이용하여 전사조절인자인 HSF1 유전자가 치환된 변이주를 구축할 수 있었다. 덧붙여, 모든 유전자에 이 기법을 적용할 수는 없다는 것을 확인하였다. 앞으로 이 변이주를 통해 기능 유전체학적인 측면에서 이 유전자의 스트레스와의 관련성을 연구하고자 한다.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.570-575
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• 2004

Three different strains of carotenoid accumulating XantlwphyUomyces dendrorhous mutants, JH1, JH2, and JH3, were isolated by NTG (N-methyl-N'-nitro-N-nitrosoguanidine) mutagenesis, which might potentially be useful for animal feed as well as for studies on the regulation and biosynthesis of astaxanthin. Mutants were selected based on the capability of growth and carotenoid production on the YM agar plate containing chemical inhibitor, $\beta$-ionone. Astaxanthin-overproducing mutant JH1 produced 4.032 mg astaxanthinlg dry cell weight, and this value was about 15-folds higher than that of wild-type. $\beta$-Carotene-overproducing mutant JH2 produced 0.273 mg $\beta$-carotene/g dry cell weight, and this was 4-folds increase from that of wild-type. In contrast, JH3 was a white-colored mutant that was unable to produce carotenoid pigment.

• 한국식품과학회지
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• 제42권5호
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• pp.549-553
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• 2010

본 연구에서는 다양한 효모 배양액을 자가소화 시킨 후 관능검사를 통해 1차적으로 8종(S. cerevisiae 3종, C. utilis 4종 및 Z.rouxii 1종)을 선별하였다. 이 중 정미성과 균체 성장율을 고려하여 C. utilis인 SEM-Y8을 선정하여 고글루타치온 생산을 위한 균주 개량을 수행하였다. 두 번의 UV 처리를 통해 SEM-Y8보다 글루타치온 생성능이 2.0배 증가된 SY8-M2-1 개량 효모 균주를 얻었으며, 선택배지(4.0% yeast extract, 1.5% glucose, 0.04% $NH_4H_2PO_4$, 0.04% L-cystein)를 사용함으로써 글루타치온 함량을 1.5배 더 향상시킬 수 있었다. 이상의 조건에서 얻어진 고글루타치온 함유 효모추출물은 마늘과 양파엑기스보다 우수한 감칠맛과 부드럽고 깊은 맛을 나타내었으며, SEM-Y8 대비 3.2배 향상된 라디컬 소거능을 나타내었다.