• BMB Reports
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• v.31 no.1
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• pp.77-82
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• 1998

To purify the geranylgeranyl protein transferase type I (GGPT-I) efficiently, a gene expression system using the pGEX-4T-1 vector was constructed. The cal1 gene, encoding the ${\beta}$ subunit of GGPT-I, was subcloned into the pGEX-4T-1 vector and co-transformed into E. coli cells harboring the ram2 gene, the ${\alpha}$ subunit gene of GGPT-I. GGPT-I was highly expressed as a fusion protein with glutathione S-transferase (GST) in E. coli, purified to homogeneity by glutathione-agarose affinity chromatography, and the GST moiety was excised by thrombin treatment. The purified yeast GGPT-I showed a dose-dependent increase in the transferase activity, and its apparent $K_m$ value for an undecapeptide fused with GST (GST-PEP) was $0.66\;{\mu}M$ and the apparent value for geranylgeranyl pyrophosphate (GGPP) was $0.071\;{\mu}M$.

• BMB Reports
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• v.34 no.5
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• pp.428-433
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• 2001

A defect in uvsC of Aspergillus nidulans caused high methyl methansulfonate (MMS)-sensitivity, hyporecombination, and a lack of UV induced mutation. The uvsC gene of Aspergillus nidulans shares a sequence similarity with the RAD51 gene of Saccharomyces cerevisiae. In this study, in vitro and in vivo tests were conducted in order to determine whether or not the UVSC protein had functional similarities to RAD51, the recombination enzyme in yeast. The purified recombinant UVSC protein, following expression in Escherichia coli, showed binding activity to single-stranded DNA (ssDNA), when both ATP and magnesium are present. In addition, ATPase activity was also demonstrated and its activity was stimulated in the presence of ssDNA. The UVSC protein that was expressed under the ADH promoter in S. cerevisiae suppressed in part the sensitivity to MMS of the rad51 null mutant. Similarly, when the uvsC cDNA was expressed from the nmt promoter, the MMS sensitivity of the rhp51 null mutant of Schizosaccharomyces pombe was partially complemented. These results indicate that the A. nidulans UVSC protein is a functional homologue of the RAD51 protein.

• Applied Biological Chemistry
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• v.47 no.3
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• pp.287-292
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• 2004

The optimum conditions for lignin peroxidase production were studied. Lignin peroxidase was produced almost exclusively in stationary culture with the optimum media composition of malt extract 1 g, yeast extract 0.4 g, glucose 0.4 g and distilled water 100 ml. Tween 80 at 0.005% concentration and veratryl alcohol at 0.4 mM were very effective inducers for lignin peroxidase production.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.37-43
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• 1993

The effect of stb locus of E. COLI IncFII plasmid NR1 on the stability of chimeric plasmids was investigated. First, we have isolated the stability locus (stb) from E. coli NR1 plasmid and then inserted into the three different vectors, pUC8, YRp17 and YEp24. By examining their stability in E. coli and yeast, we showed that the recombinant plasmids containing stb locus were resonably stable. Also, by comparing the amounts of the rDNA fragments per haploid genome with those of the plasmid fragments, we showed they copy number of recombinant plasmids was not increased. Consequently, the stb locus of E. coli IncFII plasmid NR1 stabilized the chimeric plasmids but did not affect the replication or copy number of plasmids.

• Journal of Microbiology
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• v.38 no.2
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• pp.88-92
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• 2000

A human ribosomal protein S3 (rpS3), which also functions as a DNA repair enzyme(UV endonuclease III), was expressed in a methylotrophic yeast, Pichia pastoris, and biochemically characterized. UV endonuclease activity was preiously characterized, and this activity of mammalian rpS3 was found to be non-specfic upon purification and storage. Under the Pichia expression system, the subcloned cDNA of the human rpS3 gene revealed a peptide of 42 kDa by SDS-PAGE and Western blot. The secreted form of human rpS3 rendered no endonuclease activity while the intracellular form showed UV specific endonuclease activity by the nick circle assay.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.362-368
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• 2018

Grasshoppers play vital role in the digestion of photosynthetically fixed carbons. With the aid of intestinal microflora, the grasshopper can degrade leaves constituents such as cellulose and hemicellulose. The purpose of this study was to examine cellulolytic yeast isolates from the gut of grasshoppers collected in Gyeonggi Province, South Korea. Among the yeast isolates, ON2, ON17 (two strains), and ON6 (one strain) showed positive cellulolytic activity in the CMC-plate assay. The sequence analyses of D1/D2 domains of the large subunit rDNA gene and the internal transcribed spacer (ITS) regions revealed that the strains ON2 and ON17 were most closely related to Papiliotrema aspenensis CBS $13867^T$ (100%, sequence similarity in D1/D2 domains; 99.4% sequence similarity in ITS) and strain ON6 related to Saitozyma flava (100% in D1/D2 domains; 99.0% in ITS). All these three yeast strains are capable of degrading cellulose; therefore, the members of endosymbiotic yeasts may produce their own enzymes for carbohydrate degradation and convert mobilized sugar monomers to volatile fatty acids. Thus, the endosymbiotic yeast strains ON2, ON17 (represents the genus Papilioterma) and ON6 (Saitozyma) belonging to the family Tremellomycetes, are unreported strains in Korea.

• Journal of Life Science
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• v.17 no.1 s.81
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• pp.137-142
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• 2007

For the production of natural pigments with microbe, the strains which produced monascus pigment were isolated, and then culture condition and extraction condition were investigated. These results are summarized as follows; The strain which ran produce monascus natural pigment was isolated from natural microbial sources and we made mutant of this strain with UV($235_{nm}$, 30 second) irradiation. The mutant was identified as Monascus sp. MK2-2. The optimal culture conditions were investigated optimal medium containing 0.3% rice powder, 0.2% yeast extract, 0.3% $NH_4H_2PO_4$ and $30^{\circ}C$ in a rotary shaker (120 rpm) for 5 days (initial pH 5.0), while the pigment production was determined at 24 hr intervals. The effective carbon sources were wheat flour > rice powder > fructose, and effective nitrogen sources were sodium nitrate > $KNO_3$ for production of the monascus natural pigment. The pigment capacity is good from 17 to 22 in C/N ratio. The production amount of monascus natural pigment was 0.38 g per 1 kg of rice. Also, extract of red yeast rice had anti-thrombosis activity like a degree of aspirin.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.3
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• pp.327-334
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• 2017

Water and 75% ethanol extracts were prepared from defatted green tea seeds and evaluated for their anti-yeast activities. The antimicrobial activities of defatted green tea seed extracts (DGTSEs) were tested against food-spoilage bacteria, yeasts, and molds. DGTSEs exhibited antimicrobial activities with minimum inhibitory concentrations of $39{\sim}1,250{\mu}g/mL$ against three bacteria, two molds, and all tested yeast strains. Ethanol extract showed higher antimicrobial activity than water extract. The stability of anti-yeast activities of DGTSEs was examined under different conditions of temperature, pH, and NaCl concentrations. The anti-yeast activities of DGTSEs were stable at pH 3~9, 0~20% NaCl, and $100^{\circ}C$ for 30 min. However, anti-yeast activities of DGTSEs decreased upon heating at $70^{\circ}C$ for 24 h or $121^{\circ}C$ for 15 min. DGTSEs were applied to food models to determine their inhibitory effects on yeast film formation. Water and 75% ethanol extracts were effective in preventing yeast film formation at concentrations more than 156 and $39{\mu}g/mL$ in soy sauce, 156 and $78{\mu}g/mL$ in pickle sauce, and 78 and $39{\mu}g/mL$ in kimchi, respectively.

• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.241-248
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• 2015

Tuak is a traditional alcoholic beverage, one of the most widely known in the North Sumateran region of Indonesia. It is produced by a spontaneous fermentation process through the application of one or more several kinds of wood bark or root, called raru (Xylocorpus wood bark or a variety of forest mangosteen), into the sap water of sugar palm (Arenga pinnata) for 2−3 days. In this research, yeast that are potentially useful for ethanol production was isolated from Tuak and identified. Based on analysis of D1/D2 domain sequence of LSU (large subunit) rRNA genes, those isolated yeast strains, HT4, HT5, and HT10 were identified as Candida tropicalis. Fermentation test of these C. tropicalis isolates displayed an ability to produce 6.55% (v/v) and 4.58% ethanol at 30℃ and 42℃, respectively. These results indicated C. tropicalis isolates more rapidly utilize glucose and obtain higher levels of the production of ethanol at the higher temperature of 42℃ than S. cerevisiae, a common yeast used for bioethanol fermentation.

• Microbiology and Biotechnology Letters
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• v.19 no.2
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• pp.193-199
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• 1991

The effects of pH on the cell growth, the elaboration of pullulan, and the morphology of Aureobasidium pullulans IFO 4464 were examined. A. pulluans grew in yeast-like form at constant pH7.5 and in mycelial form at constant pH2.5. At the both pH conditions, the elaboration of pullulan was very low, about 6.0~6.5g/l. The mixture of yeast-like form and mycelial form of cells was found at the constant pH4.5, at which condition, the elaboration of pullulan was high, about 24.5g/l. The pH shift experimemts showed that the specific production rates of pullulan were 0.048($hr^{-1}$)for the mycelial form and 0.058($hr^{-1}$)for the yeast like form, which indicated that the yeast-like form has the similar, only slightly higher, biosynthetic activity of pullulan to the mycelial form at pH4.5 and the pH of culture broth is more important factor for the elaboration of pullulan than the morphology of A. pullulans.