• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.296-305
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• 2020

Tricholoma matsutake is an ectomycorrhizal fungus, related with the host of Pinus densiflora. Most of studies on T. matsutake have focused on mycelial growth, genes and genomics, phylogenetics, symbiosis, and immune activity of this strain. T. matsutake is known for its unique fragrance in Eastern Asia. The most major component of its scent is (R)-(-)-1-octen-3-ol and is biosynthesized from the substrate linoleic acid by the sequential reaction of lipoxygenase and peroxide lyase. Here, we report for the first time the biosynthesis of (R)-(-)-1-octen-3-ol of T. matsutake using the yeast Saccharomyces cerevisiae as a host. In this study, cDNA genes correlated with these reactions were cloned from T. matsutake, and expression studies of theses genes were carried out in the yeast Saccharomyces cerevisiae. The product of these genes expression study was carried out with Western blotting. The biosynthesis of (R)-(-)-1-octen-3-ol of T. matsutake in recombinant Saccharomyces cerevisiae was subsequently identified with GC-MS chromatography analysis. The biosynthesis of (R)-(-)-1-octen-3-ol with S. cerevisiae represents a significant step forward.

• Applied Biological Chemistry
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• 제34권1호
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• pp.67-74
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• 1991

내당 내알콜성 균주인 S. cervisiae D1을 이용하여 알콜 발효를 할 때 ethanol 생산성을 증대시키기 위하여 발효시 $Ca^{2+}$을 첨가하여 발효에 미치는 영향에 관해서 연구한 결과 다음의 결론을 얻었다. 초기 glucose 농도를 400g/l로 하였을 때 $Ca^{2+}$에 의해 ethanol 생산성이 증가하였고 생균수도 증가하였으며 최적 첨가 농도는 0.8 mM이었다. 초기 glucose농도를 달리하고 발효하였을 때는 $Ca^{2+}$의 영향이 초기 glucose 농도가 높을수록 켰다. Ethanol 첨가에 의해 비증식 속도가 감소하였지만 $Ca^{2+}$을 0.8 mM 첨가시 비증식 속도의 저해가 감소되었다. 또한 ethanol을 첨가하지 많았을 때는 $Ca^{2+}$을 첨가한 경우와 첨가하지 않은 경우의 비증식 속도가 같았다. Ethanol 첨가 농도에 의해 사멸 속도가 증가하였으나 $Ca^{2+}$을 0.8 mM 첨가시 비사멸 속도가 감소하였다. Acidification curve에서도 ethanol 첨가시 final pH가 증가하였으나 $Ca^{2+}$을 0.8 mM 첨가하면 final pH가 감소하였다. 따라서 $Ca^{2+}$ 첨가시 ethanol에 대한 내성이 더 증가된다는 것을 알 수 있었다.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1986년도 추계학술대회
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• pp.505.1-505
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• 1986

The gene for iso-1-cytochrome c for Saccharomyces cerevisiae was recloned into a pSP65 vector containing an active bacteriophage SP6 promoter. The iso-1-cytochrome c gene was cloned as an 856 bp Xho 1-Hind III fragment. When the resulting plasmid was digested at the Hind 111 site 279 bases downstream from the termination codon of the gene and transcribed in vitro using SP6 RNA polymerase, full length transcripts were produced. The SP6 iso-1-cytochrome c mRNA was translated using a rabbit reticulocyte lysate system and the protein products analyzed on SDS polyacrylamide gels. One major band was detected by autofluorography. This band was found to have a molecular weight of 12,000 Da and coincided with the Coomassie staining band of apocytochrome c from S. cerebisiae. The product was also shown to be identical with that of standard yeast apocytochrome c on an isoelectric focusing gel. The in vitro synthesized iso-a-cytochrome c was methylated by adding partially purified S-adenosyl-L-methionine . protein-lysine N-methyltransferase (Protein methylase III; EC 2.1.1.43) from S. cerevisiae along with S-adenosyl-L-methionine to the in vitro translation mixtures. The methylation was shown to be inhibited by the addition of the methylase inhibitor S-adenosyl-L-homocysteine or the protein synthesis inhibitor pu omycin. The methyl derivatives in the protein were identified as $\varepsilon$-N-mono, di and trimethyllysine by amino acid analysis. The molar ratio of methyl groups incorporated to that of cytochrome c molecules synthesized showed that 23% of the translated cytochrome c molecules were methylated by protein methylase III.

• Journal of Microbiology and Biotechnology
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• 제20권10호
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• pp.1397-1402
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• 2010

Papyriflavonol A (PapA), a prenylated flavonoid [5,7,3',4'-tetrahydroxy-6,5'-di-(${\gamma},{\gamma}$-dimethylallyl)-flavonol], was isolated from the root barks of Broussonetia papyrifera. Our previous study showed that PapA has a broad-spectrum antimicrobial activity against pathogenic bacteria and fungi. In this study, the mode of action of PapA against Candida albicans was investigated to evaluate PapA as an antifungal agent. The minimal inhibitory concentration (MIC) values were 10~25 ${\mu}g/ml$ for C. albicans and Saccharomyces cerevisiae, Gram-negative bacteria (Escherichia coli and Salmonella typhimurium), and Gram-positive bacteria (Staphylococcus epidermidis and Staphylococcus aureus). The kinetics of cell growth inhibition, scanning electron microscopy, and measurement of plasma membrane florescence anisotrophy revealed that the antifungal activity of PapA against C. albicans and S. cerevisiae is mediated by its ability to disrupt the cell membrane integrity. Compared with amphotericin B, a cell-membrane-disrupting polyene antibiotic, the hemolytic toxicity of PapA was negligible. At 10~25 ${\mu}g/ml$ of MIC levels for the tested strains, the hemolysis ratio of human erythrocytes was less than 5%. Our results suggest that PapA could be a therapeutic fungicidal agent having potential as a broad spectrum antimicrobial agent.

• Journal of Pharmaceutical Investigation
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• 제10권4호
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• pp.8-22
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• 1980

In order obtain some new antibacterial agents, seven new 2-ethoxymethyl-3-(5-nitro-2-furyl) acrylamide derivatives were synthesized by condensing 2-ethoxymethyl-3-(5-nitro-2-furyl) acyloyl chloride with amino compounds namely 5-amino-3, 4-dimethyl isoxazole, sulfamonomethoxazole, d-2-amino-1-butanol, hydroxylamine hydrochloride, semicarbazide hydrochloride, thiosemicarbazide, and p, p'-diaminodiphenylsulfone, respectively. The seven synthesized compounds were 2-ethoxymethyl-3-(5-nitro-2-furyl) acryl-5-amino-3, 4-dimethylisoxazoleamide [VII], $N^4-[2-ethoxymethyl\;3-methyl\;(5-nitro-2-furyl)\;acryl]-N^1-(5-methyl-3-isoxazolyl)$ sulfanilamide [VIII], 2-ethoxyl-3-(5-nitro-2-furyl) acrylsemicarbazide [X], 2-ethoxymethyl-3-(5-nitro-2-furyl) acrylthiosemicarbazide [XI], 2-ethoxymethyl-3-(5-nitro-2-furyl) acryl-d-2-amino-1-butanolamide [XII], and 4, 4'-di[2-ethoxymethyl-3-(5-nitro-2-furyl) acryl-amido] diphenylsulfone [XIII]. These compounds, with exception of the compound XIII, showed generally effective antibacterial activity, especially in the following instances. Compound VII was shown to be effective against Bacillus subtilis ATCC 6633 compound VIII, against Bacillus cereus var. Mycoides ATCC 1778, and compound XII, against both Proteus vuglaris and Saccharomyces cerevisiae ATCC 9763.

• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.293-299
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• 2000

Many methods have been developed for screening chemicals with hormonal activity. Using recombinant yeasts expressing either human estrogen receptor [Saccharomyces cerevisiae ER + LYS 8127 (YER)] or androgen receptor [S. cerevisiae AR + 8320 (YAR)], we evaluated the hormonal activities of several chemicals by induction of ${\beta}-galactosidase$ activity. The chemicals were $17{\beta}-estradiol$ (E2), testosterone (T), ${\rho}-nonylphenol$ (NP), bisphenol A (BPA), genistein (GEN), 2-bromopropane (2-BP), dibutyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and butylparaben (BP). To assess the estrogenicity of NP, the result of the in vitro recombinant yeast assay was compared with an E-screen assay using MCF-7 human breast cancer cells and an uterotrophid assay using ovariectomized mice. In the YER yeast cells, E2, NP, BPA, GEN, and BP exhibited estrogenicity in a doseresponse manner, while TCDD did not. All the chemicals tested, except T, did not show androgenicity in the YAR yeast cell. The sensitivity of the yeast (YER) assay system to the estrogenic effect of NP was similar to that of the E-screen assay. NP was also estrogenic in the uterotrophic assay. However, in terms of convenience and costs, the yeast assay was superior to the E-screen assay or uterotrophic assay. These results suggest that the recombinant yeast assay can be used as a rapid tool for detecting chemicals with hormonal activities.

• 미생물학회지
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• 제52권4호
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• pp.487-494
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• 2016

출아 효모에서의 Paf1 복합체는 총5개의 단백질로 구성되어있고, 구성성분들은 출아효모, 초파리, 식물들, 그리고 인간에 이르기까지 구조적으로, 기능적으로 잘 보존되어 있다. RNA 중합효소 II와 결합한 상태로 전사 개시부위부터 종결부위까지 함께 이동하며, 여러 전사인자들의 유입을 위한 매개체로 작용하여, 유전자 발현 조절의 핵심적인 역할을 수행한다. Paf1 복합체는 H2BK123 monoubiquitination에 기여하고, histone crosstalk에 의해 간접적으로 H3K4의 di-, tri-methylation에 기여하는 것이 알려져 있다. 하지지만, Paf1 복합체 구성요소들의 개별적인 기능에 대해서는 연구가 되어있지 않다. 이 연구에서는, Paf1 복합체 구성요소들의 단일 결핍 돌연변이 균주를 만든 후, 이들의 H2BK123 monoubiquitination 및 H3K4 mono-, di-, tri-methylation에 미치는 영향을 관찰했다. 놀랍게도, ${\Delta}paf1$, ${\Delta}rtf1$, ${\Delta}ctr9$ 돌연변이 균주에서는 H2Bub에 영향을 받는 H3K4me2와 H3K4me3뿐 아니라, H2B monoubiquitination에 영향을 받지 않는 H3K4 monomethylation의 심각한 감소를 관찰했다. 그러나, methyl기 전달 효소인 Set1의 발현 정도는 이 돌연변이 균주들에서 변하지 않았다. 이러한 결과로부터, Paf1 복합체가 Set1의 활성이나 Set1 복합체의 안정성을 직접 조절함으로써 H3K4 methylation을 조절할 수 있음을 제시한다.