• The Korean Journal of Mycology
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• v.23 no.2 s.73
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• pp.129-138
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• 1995

DNA fragment being able to restore in vitro activity of ${\beta}-1,3-glucan$ synthase was cloned by transformation of the Saccharomyces cerevisiae LP353 mutant strain with genomic library constructed in the YCp50. For the selection of transformants which showed no detectable phenotype linked to recovery of the defect in ${\beta}-1,3-glucan$ synthase activity, the colony autoradiography was succesfully applied. The restriction map of the cloned DNA fragment, which is 8.5-kb in length, was constructed. Both the YEplac195 and the YCp50 carrying the 8.5-kb fragment increased ${\beta}-1,3-glucan$ synthase activity of LP353 by two fold. Neither the YEplac195 nor the YCp50 carrying the 8.5-kb DNA fragment, however, complemented the temperature-dependent osmotic sensitivity which is another distinctive phenotype of LP353. Subcloning experiments indicated that a functional region was located in 4.8-kb BglII-KpnI fragment. The 4.8-kb fragment was also able to increase the level of ${\beta}-1,3-glucan$ content in cell wall as well as the resistance of cells to cell wall lytic enzyme, ${\beta}-1,3-glucanase$. The growth rate of the LP353 with 4.8-kb fragment was almost same as that of wild type strain in liquid medium with 1.2 M sorbitol at nonpermissive temperature. Taken these results together, the 4.8-kb fragment seemed to contain the BGS2 gene for ${\beta}-1,3-glucan$ synthase activity in yeast S. cerevisiae.

• Journal of Life Science
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• v.7 no.1
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• pp.30-38
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• 1997

For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

• Microbiology and Biotechnology Letters
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• v.45 no.1
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• pp.81-86
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• 2017

Alignment of 967 reference sequences of the typical 2-Cys peroxiredoxin family of proteins revealed that 10 amino acids were conserved, with over 99% identity. To investigate whether the conserved aspartic acid residues and serine residue affect the peroxidase and chaperone activity of the protein, we prepared yeast TSA1 mutant proteins in which aspartic acids at positions 75 and 103 were replaced by valine or asparagine, and serine at position 73 was replaced by alanine. By non-reducing SDS-PAGE, TSA1 and the S73A, D75V and D75N mutants were detected in dimeric form, whereas the D103V and D103N mutants were detected in various forms, ranging from high molecular-weight to monomeric. Compared with wild type TSA1, the D75N mutant exhibited 50% thioredoxin peroxidase activity, and the S73A and D75V mutants showed 25% activity. However, the D103V and D103N mutants showed no peroxidase activity. All proteins, except for the D103V and D103N mutants, exhibited chaperone activity at $43^{\circ}C$. Our results suggest that the two conserved aspartic acid residues and serine residue of TSA1 play important roles in its thioredoxin peroxidase activity, and D103 plays a critical role in its chaperone activity.

• Korean Journal of Food Science and Technology
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• v.29 no.3
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• pp.588-594
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• 1997

Yeast strains useful for the production of wine using mandarine orange, Citrus unshiu, as a main substrate were screened, and their primary ability to decompose citric acid that affects directly wine quality was investigated. Total eleven strains were selected for brewing orange wine. Five wild strains were from soil-based collections and identified: four of them were Saccharomyces cerevisiae and one of them was S. ellipsoideus. The rest of six strains were from among eighteen laboratory strains: three of them were S. cerevisiae, and the other three were S. coreanus, S. uvarum, and S. sake. Two strains of S. cerevisiae out of these selections were chosen and their decomposition of citric acid was investigated. Citric acid was not utilized as sole carbon source for cellular growth. However, when both citric acid and glucose were added together as carbon sources, decrease of citric acid concentration was observed after incubation. Shaking incubation was more effective for the reduction of citric acid than standing incubation. Utilization of citric acid did not contribute to the increase of ethanol concentration during fermentation. On the other hand, it appeared that citric acid caused partial inhibition of cellular growth of the yeasts.

• KSBB Journal
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• v.15 no.6
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• pp.566-572
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• 2000

Experimental studies were carried out to develop an osmotolerant mutant of Candida magnoliae JH and to determine the optimum concentrations of carbon and nitrogen sources to improve erythritol yield and productivity. Mutants of C. magnoliae JH were prepared by treatment with ethylmethane sulfonate, and osmotolerant mutants were isolated from the agar plate colonies containing 2.5 M KCI. Among the mutants isolated, one mutant M26 was finally selected based on erythritol yield and productivity. In shake flask culture, glucose proved to be the best carbon source and the optimum yeast extract concentration was 5 g/L based on 100 g/L glucose. The erythritol yield and productivity of mutant M26 were greater than wild type in 100 g/L glucose medium. in the fermentation experiments, erythritol production increased with increased glucose concentration, up to a limit of 250 g/L. The maximum concentration of erythritol achieved 127.5 g/L, and the yield and productivity of erythritol production were 51.0% and 0.63 g/L-h, respectively.

• Korean Journal of Food Science and Technology
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• v.37 no.4
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• pp.574-578
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• 2005

Effects of different yeast strains on physicochemical characteristics of Bokbunja (Rubus coreanus Miq.) fruits alcohol fermentation were investigated. Bokbusnja fruit must was inoculated with Saccharomyces cerevisiae KCCM 12224 (Sc-24), wild-type Bokbunja yeast (Bok-3), Saccharomyces coreanus (Yak-7), and Sc-24+Yak-7. Ethanol contents of Sc-24, Bok-3, Yak-7, and Sc-24+Yak-7 were 11.08, 10.62, 10.18, and 10.26%, respectively after 10 days fermentation. Addition of pectinase (500 ppm) increased ethanol content by 0.1-1.5%. Organic acids of Bokbunja wine were citric, malic, shikimic, formic, and oxalic acids. Citric and malic acid contents remarkably decreased, whereas that of acid increased by fermentation. Total acidity of Bokbunja wine was dependent on citric acid content. Sc-24, Yak-7, and Bok-3+ pectinase were more efficient for improvement of wine-color, although color values of Bokbunja wine significantly decreased during early stage of fermentation. Sc-24 and Bok-3+500 ppm of pectinase, and 8-10 days of fermentation could enhance quality of Bokbunja wine.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.102-108
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• 1994

Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

• Journal of Life Science
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• v.27 no.3
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• pp.339-345
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• 2017

Carotenoids are natural lipid-soluble pigments, which are produced primarily by bacteria, algae, and plants. Many studies have focused on the identification, production, and utilization of natural sources of astaxanthin from algae, yeast, and crustacean byproducts as an alternative to the synthetic pigment, which is mostly used today. The aim of the present study was to identify a mutant of Paracoccus haeundaensis by exposure to UV and ethyl methanesulfonate (EMS). The mutant was then exposed to nutrient stress conditions to isolate an astaxanthin-hyperproducing strain, followed by characterization of the mutant. The survival rate decreased in accordance with an increase in the UV exposure time and an increase in the EMS concentration. A mutant of the original P. haeundaensis strain was identified that showed hyperproduction of astaxanthin following exposure to UV irradiation (20 min) and EMS treatment (0.4 M concentration). The optimal culture conditions for the PUE mutant were $25^{\circ}C$, pH 7-8, and 3% NaCl. The effects of various carbon and nitrogen sources on the growth and astaxanthin production of PUE were examined. The addition of 1% raffinose and 3% potassium nitrate influenced cell growth and astaxanthin production. The selected mutant exhibited an increase of 1.58 folds in astaxanthin content compared to initial wild type strain. A genetically stable mutant strain obtained using mutagen (UV irradiation and EMS treatment) may be a suitable candidate for further industrial scale production of astaxanthin.

• KSBB Journal
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• v.21 no.4
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• pp.286-291
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• 2006

Various ${\beta}$-ionone resistant mutants were isolated from the wild-type red yeast Xanthophyllomyces dendrorhous KCTC 7704. Although the growth of X. dendrorhous KCTC 7704 was strongly inhibited at 0.025 mM ${\beta}$-ionone, one of the ${\beta}$-ionone resistant mutants isolated at 0.1 mM ${\beta}$-ionone by NTG mutagenesis showed rather 70% of relative survival at 0.15 mM ${\beta}$-ionone. Fermentation kinetics study with the mutant was carried out at $20^{\circ}C$ for 4 days in 300-mL baffled flasks. The mutant yielded up to 2.3-fold higher carotenoids content(viz. $1.2{\mu}g$ of total carotenoids per mg of dry cells) compared with the wild-type strain. The production of metabolites such as organic acids could be neglected. Studies on the kinetics with various carbon substrates revealed both an increase in final dry cell mass and a higher total carotenoids content in cell mass with glucose when compared to fructose or sucrose. As a further part of study, the effect of pH on the fermentation kinetics was investigated in glucose-limited chemostat at a dilution rate of $0.04h^{-1}$. When compared to steady-state kinetic parameters obtained at pH 4.0, a significant reduction in cell concentration at pH 3.0 and a lower carotenoids content at pH 5.2 were evident.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.2
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• pp.148-157
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• 2005

This study was investigated to determine antimicrobial activity of the water and ethanol extracts of Korean wild green tea, semi-fermented tea, and fermented tea. Antimicrobial activity was examined against 8 kinds of several microorganisms. The minimum inhibitory concentration (MIC) of the water and ethanol extracts of green tea showed the most active antimicrobial activity against B. subtilis 0.2 mg/mL in Gram positive bacteria and P. fluorescens 0.3∼0.5 mg/mL in Gram negative bacteria. But the extracts did not show antimicrobial activity against lactic acid bacteria and yeast at the level of less than 1 mg/mL. Antimicrobial activity got lower as tea got more fermented. Antimicrobial activity of ethanol extracts from green tea, semifermented tea, and fermented tea was stronger than that of water extracts. Antimicrobial activity of the water and ethanol extracts of green tea, semi-fermented tea, and fermented tea was not destroyed at 50∼121$^{\circ}C$, and pH 3∼11, which proved to be very stable when given over heat, acid & alkali treatment. The ethanol extract of green tea, semi-fermented tea, and fermented tea was fractionated in the order of hexane, diethyl ether, ethyl acetate and water fraction. The highest antimicrobial activity was found in the water fraction, but not found in hexane fraction, while antimicrobial activity of fermented tea was not found in ether fraction.