• Journal of Life Science
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• v.26 no.1
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• pp.17-22
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• 2016

Ethanol is a very valuable material, however, it is also a source of stress, as the accumulation of ethanol in a medium inhibits cell viability and decreases productivity of the target product. Therefore, the ethanol tolerance of yeast, which is closely related to ethanol productivity, is an important factor in industrial ethanol production. In this study, the YDJ1 and PEP5 genes were selected as target genes for elucidating ethanol-tolerant mechanisms by analyzing the expression regulation of these genes. The pA-YDJ1 and pA-PEP5 plasmids containing YDJ1 and PEP5 genes under an ADH1 promoter, respectively, were constructed and transformed into BY4742 (host strain), BY4742△ydj1, and BY4742△pep5 strains. The ethanol tolerance in the BY4742△ydj1/ pA-YDJ1 and BY4742△pep5/pA-PEP5 transformants was restored by overexpression of the YDJ1 and PEP5 genes to the host strain level. The YDJ1 and PEP5 genes were also introduced into the double gene disruptant (BY4742△ydj1△pep5) to investigate the expression regulation of the YDJ1 and PEP5 genes. The simultaneous overexpression of the YDJ1 and PEP5 genes restored ethanol tolerance to the 90% level of the BY4742 strain under 8% ethanol stress. The YDJ1 gene induced more overexpression of the PEP5 gene in the BY4742△ydj1 △pep5/pA-YDJ1, pA-PEP5 strain, suggesting that the YDJ1 gene partially regulates the expression of the PEP5 gene as an upstream regulator.

• Korean Journal of Food Science and Technology
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• v.31 no.2
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• pp.465-474
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• 1999

RNase activity of Saccharomyces cerevisiae ATCC 7754 was investigated to obtain strains with high ribonucleic acid (RNA) content. The yeast strain contained two RNase activities; an acidic RNase with a optima of pH $3{\sim}4$ and an alkaline RNase with a optima pH 9. The acidic RNase activity was inhibited by $0.08\;M\;HgCl_{2}$ most drastically. The alkaline RNase activity was inhibited by 2.0 M NaCl or KCl, while enhanced by addition of $0.05\;M\;CaCl_{2},\;0.02\;M\;ZnSO_{4},\;or\;0.008\;M\;HgCl_{2}$. Various mutants of Saccharomyces cerevisiae ATCC 7754 were isolated by ethylmethane sulfonate (EMS) treatment or $\gamma$-ray/ultra violet irradiation. Among the mutants that were sensitive to high concentration of KCl which inhibits alkaline RNase, B24 was selected for high RNA content per culture volume. Growth characteristics of the mutant were comparable to those of the mother strain with optimum growth at pH $4.5{\sim}5.5$. The mutant accumulated higher content of RNA than the mother strain when glucose was used as the carbon source. However, both growth rate and total RNA content of the mutant were higher in molasses medium than in glucose medium. RNA content of the mutant increased rapidly during the early stage of growth, and then decreased gradually until the culture reached stationary phase by a fed-batch culture in a 5 L jar fermenter. Maximal cell harvest and the final RNA content using the mutant B24 were 69.6 g/L culture broth and 19.8 g/100 g of the dry cell while those using the mother strain were 68 g/L culture broth and 16.1 g/100 g of dry cell, respectively.

• Journal of Mushroom
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• v.2 no.2
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• pp.88-96
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• 2004

This study was executed to decide the physiological characteristics of Ferule mushroom. Four strains of Ferule mushroom were tested to select a superior strain in its mycelial growth. The pertinent substrates, temperature and pH ranges for the growth of selected strain were determined. And then, the wood rotting ability and type of the Ferule mushroom were determined. The superior strain F-2 among four strains was selected, on the basis of its vegetative mycelial growth and density on agar media. Mycelial growth of F-2 was the best on MYPA among other tested synthetic or semi-synthetic media. The temperature range for pertinent mycelial growth was about $25{\sim}34^{\circ}C$ and best at $30^{\circ}C$. The optimum pH range on MYPA was 5.0~6.0. The mycelial growth was mostly stimulated by soluble starch at cont. 1% (w/w) and secondly, maltose among several carabon sources and by mixed solution of YE(0.25%) and ME(0.25%) but not by ME alone. Cell thining and erosion of Pinus rigida wood by the mycelia of Ferule mushroom were found only on a few cell but largely at wood block test, indicating that the softwood rotting ability of Ferule mushroom mycelia was not so good. The result of polarized light microscopy appeared that cellulose of some tracheides showing the S3 layer lost brifringence was degraded by Ferule mushroom. But only part of cellulose of P. rigida wood was degraded by Ferule mushroom, because most of wood cells continued to showing briefingence. A largely degraded ray parenchyma and longitudinal parenchyma cell and partly thinning and erosion of hardwood(Quercus serrata) cell was found and it indicates that the rotting ability of Ferule mushroom mycelia on hardwood was higher than on softwood. It could be concluded that the difference in the wood rot by Ferule mushroom between the hardwood and softwood was made by the difference of chemical constitutions between them, especially in the contents and the types of lignin. Ferule mushroom was considered as white rotter as a result of bavendam test, although more research should be required.

• Food Science and Preservation
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• v.18 no.4
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• pp.559-566
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• 2011

In this study, the good brewing conditions for the 24 $^{\circ}Brix$ freeze-concentrated apple wine were investigated. The four selected Saccharomyces cerevisiae strains MM10, SS89, SS812, and WW108, could ferment quickly when brewed with high sugar levels. During the fermentation, the reducing sugar contents slowly declined while the total acid in all the yeasts increased and the final alcohol content was 12-13%, a typical wine's alcohol content. The viable counts were shown to be 6-6.8 log cfu/ml. During the fermentation, the organic acid content was shown to be within the range of 2.36-3.11%, and the free sugar content, except for the SS89 and WW108 strains, was shown to consist only of sorbitol, although fructose was somewhat detected in the SS89 and WW108 strains. Methanol was not detected, or only a trace of it was detected, and the aldehyde content was 107.68-114.27 ppm. As for the fusel oil contents, a trace of propanol was detected. Isobutanol and butanol were present in 40.16-54.65 and 25.47-27.73 ppm, respectively. The isoamy1 alcohol content was shown to be the highest (108.88-217.26 ppm). The final total phenolic compounds were shown to be 0.1-0.16%. The final Hue values were shown to be 1.3-3.6, and the final intensity was 0.1-0.45. The lightness (L) was within the range of 91.78-98.51, the redness (a) was at a neutral position at red and green, and the yellowness (b) was within the range of 2.38-7.7. In the sensory evaluation, the SS812 strain was found to be the best in terms of color, the SS89 strain in terms of odor, and the WW108 strain in terms of taste. Overall, SS812 was found to be the best apple wine.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.6
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• pp.695-702
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• 2017

This study was carried out to investigate the effects of seed vinegar on antioxidant activity and antimicrobial activities of concentrated blueberry vinegar (CBV). Of the nine strains of yeast and six strains of acetic acid bacteria provided by the Microbial Institute for Fermentation Industry, each strain of yeast (Saccharomyces cerevisiae SRCM 100610, showing the highest ethanol content) and acetic acid bacteria (Acetobacter pasteurianus SRCM 101342, showing the highest total acidity) was selected for production of CBVs. Sugar content, pH, total acidity, total phenolic content (TPC), and browning intensity (280 nm and 420 nm) in CBVs using concentrated blueberry juice were $11.05{\sim}12.70^{\circ}Brix$, 2.63~2.98, 1.65~5.72%, 3.03~4.24 mg/mL, 0.95~1.50, and 0.11~0.20, respectively. Sugar content and total acidity of CBVs increased upon addition of seed vinegar, whereas pH, TPC, and browning intensity decreased. Of all CBVs with various additions of seed vinegar, the control showed the lowest $EC_{50}$ values in DPPH radical scavenging assay, ABTS radical scavenging assay, and reducing power (23.80, 19.48, and 79.21 dilution factor, respectively), whereas the 40% seed vinegar group showed the highest clear zone diameter values for Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus (4.31, 4.59, 5.81, and 3.97, respectively). Antioxidant activities of CBVs were closely correlated with their TPC, browning intensity at 280 nm, pH, and total acidity values, showing correlation determination coefficient ($R^2$) values higher 0.82. However, antimicrobial activities of CBVs were closely correlated with their pH and total acidity values, showing higher $R^2$ values more than 0.92. These results suggest that CBVs using concentrated blueberry juice, S. cerevisiae SRCM 100610, and A. pasteurianus SRCM 101342 may be useful as potentially functional foods for enhancing health.

• Korean Journal of Food Science and Technology
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• v.41 no.5
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• pp.541-546
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• 2009

The objectives of this study were to evaluate the physico-chemical and sensory characteristics of a Korean traditional alcoholic beverage (yakju) prepared using different nuruk (Korean-style koji) concentrations and yeasts such as the fusant FA776 and Saccharomyces cerevisiae KOY-1, respectively. The fusant FA776, which has alcohol-fermenting and starch-utilizing properties, was formed by Saccharomyces cerevisiae KOY-1 and Saccharomyces diastaticus KCTC1804. The fermentation trial was conducted in a 5 L lab-scale jar at $25^{\circ}C$. The maximum alcohol production of the K-100 and F-50 reached levels of 135.0 mg/mL and 119.4 mg/mL, respectively. The pH values were in a range of 4.3-4.5. Total acidity was in a range of 0.47-0.60%. Organic acids and amino acids were analyzed in order to evaluate variations in its composition and content via HPLC analysis. Organic acids including lactic acid, citric acid, malic acid, and pyruvic acid, and 16 kinds of amino acids, including aspartic acid, were detected in all treatments. K-100 showed the highest amino acid contents, whereas F-50 exhibited the lowest amino acid contents. Volatile flavor components such as phenylethyl alcohol, isoamyl alcohol, 2-methylthiophane, isobutyl alcohol, and ethyl succinate were detected as a major component in all treatments, as determined via gas chromatography. The results of our sensory evaluation demonstrated that Yakju fermented by the FA776 fusant yielded more favorable results than S. cerevisiae KOY-1.

• KSBB Journal
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• v.22 no.5
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• pp.297-304
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• 2007

For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.201-209
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• 2014

This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $KH_2PO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, and 0.15% yeast extract at $30^{\circ}C$. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.

• KSBB Journal
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• v.15 no.2
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• pp.125-133
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• 2000

Specific carotenoids and astaxanthin biosynthesis power of Phaffia rhodozyma mutant 876, which was obtained after NTG a and UV treatments, was higher than those of the wild type by 40% and 50%, respectively. The mutant strain did not show t the catabolite repression even at 22% (w/v) glucose concentration. The optimum C{N ratio was 2.0, and the optimum t temperature and initial pH were $22^{\circ}C$ and 6.0, respectively. 80th cell growth and astaxanthin formation decreased drastically a as the fermentation temperature was increased over $22^{\circ}C$, whereas they were comparable in the pH range between 5.0 and 7 7.0. Inoculum size did not affect the final cell density nor the carotenoids biosynthesis, and 3%(v/v) was selected as optimal. H Higher dissolved oxygen concentration facilitated astaxanthin biosynthesis, and aeration rate of 1.0 v/0/m and agitation speed of 400 rpm were selected as optimum. The final cell dens때 of 43.3 g/L and the volumetric astaxanthin and carotenoids concentrations of 110.6 mg/L and 149.4 mg/L, respectively, were obtained. The specific carotenoids concentration was 3.45 m mg{g-yeast(dry). Yx/s and Yp/s values of 0.37 and 1.08 were obtained. The result of this study will provide basic information u useful for mass production of astaxanthin from P. rhodozyma fermentation.

• Korean Journal of Microbiology
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• v.39 no.4
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• pp.267-271
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• 2003

Burkholderia sp. D5, a polyaromatic hydrocarbons(PAHs)-degrading bacterium, was isolated from oil-contaminated soil. The bacterium could utilize phenanthrene (Phe) as a sole carbon source but could not use pyrene (Pyr). However, the strain could degrade Pyr when a cosubstrate such as yeast extract (YE) was supplemented. The PAH degradation rate of the bacterium was enhanced by the addition of other organic materials such as YE, peptone and glucose. YE was a particularly effective additive in stimulating cell growth as well as PAH degradation. When 1 g-YE/L was supplemented into the basal salt medium (BSM) with 215 mg-Phe/L, the specific growth rate (0.28 h-1) and Phe-degrading rate (29.30 μmol/L/h) were enhanced approximately ten and two times more than those obtained in the BSM with 215 mg-Phe/L, respectively. Through kinetic analysis, the maximum specific growth rate (μmax) and PAH degrading rate (Vmax) for Phe were obtained as 0.34/h and 289 ${\mu}mol$/L/h, respectively. Also, μmax and Vmax for Pyr were 0.27 h-1 and 50 ${\mu}mol$/L/h, respectively. The degradation rates for each Phe (2.20 μmol/L/h) and Pyr (2.18 μmol/L/h) were lower in mixture substrates than in a single substrate (29.30 ${\mu}mol$/L/h and 9.58 ${\mu}mol$/L/h, respectively). Burkholderia sp. D5 can degrade Phe and Pyr contained in soil, and the PAH degradation rates in soil were 20.03 ${\mu}mol$/L/h for Phe and 1.09 ${\mu}mol$/L/h for Pyr.