• Mycobiology
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• v.31 no.4
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• pp.191-195
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• 2003

Ethanol treatment method was attempted for the selective isolation of ethanol-tolerant fungi from two sites of rice paddy fields around Seoul area. The vertical and seasonal fluctuation of the fungal population were also investigated. The ethanol-tolerant fungi were Talaromyces stipitatus, T. flavus var. flavus, T. helicus var. major, Eupenicillium javanicum, Emericellopsis terricolor, Pseudourotium zonatum, Aspergillus flavus, Cladosporium cladosporioides, Penicillium frequentans, P. janthinellum, and P. verruculosum. The most dominant species isolated by this method was T. stipitatus. It was found that the numbers of fungal species and colony forming units(CFUs) of ethanol-tolerant fungi were higher in Ascomycota than in Deuteromycota. A particular tendency appeared the highest CFUs in autumn, but lower in spring and winter. T. stipitatus was the dominant species of ethanol tolerant microfungi. This result would suggest that membrane lipid composition of ethanol-tolerant fungi isolated from the soils may play on important role in the ethanol tolerance.

• Korean Journal of Microbiology
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• v.29 no.2
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• pp.136-142
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• 1991

The selection of ethanol-tolerant strains was applied to enrichment culture of YPD broth medium containing various concentrations of ethanol. Isolates were identified to be Saccharomyces cerevisiae, the others as S. dairensis, S. exiguus, S. telluris, Saccharomycodes ludwigii, Schwanniomyces occidentalis var. occidentalis and Zygosaccharomyces florentinus. Among isolates S. cerevisiae YO-1 was screened as having the highest ethanol tolerance and produced 18% (v/v) ethanol after 4 days fermentation. The change of fatty-acyl residues represents that a progressive decrease in fatty-acyl unsaturation and a proportional increase in saturation in phospholipids of yeast cells during fermentation affected the yeast viability. Supplementation ethanol to the cultures led to an increase of unsaturated fatty-acyl residues, especially $C_{16}$ or $C_{18}$ residues, along with a decrease in the proportion of saturated residues in cellular phospholipids. Increasing the amount of soy flour led to an increase in the maximum number of viable yeast cells and ethanol production. It was possible in 4 days to reach 21% (v/v) ethanol by adding 4% soy flour as source of unsaturated fatty-acyl residues to the fermentation medium. Soy flour not only increased yeast population but also enhanced the physiological properties of yeast cells to be ethanol tolerant in the anaerobic culture.

• Journal of Microbiology and Biotechnology
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• v.2 no.3
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• pp.226-229
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• 1992

An ethanol tolerant mutant was selected by successive transfers of Clostridium thermohydrosulfuricum ATCC 33223 into the media with progressively higher ethanol concentrations. The growth kinetics of the mutant were characterized under various growth conditions. Physiological differences such as enhanced growth, tolerance to various solvents, alteration of the optimum temperature and the ratio of end products during fermentation were noticed in the mutant.

• Microbiology and Biotechnology Letters
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• v.30 no.3
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• pp.216-222
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• 2002

To Produce the modified Cheongiu that has high ethanol content, an ethanol-tolerant strain Saccharo-myces cerevislae SE2l1 was screened from Saccharomyces cerevisiae Kyokai No. 10 strain. The isolate showed faster growth than in the medium containing 10% ethanol compared with original strain. The isolate produced a higher concentration of ethanol and showed higher resistance to ethanol, high osmolarity and heat than the original strain. The analyses of yeast membrane components indicated that there were no significant changes in composition of sterols and phospholipids between the isolated and the original strain. However, during the fermentation, the iso-lated strain could change the fatty acid composition in the membrane more rapidly in the direction of decreasing membrane unsaturation and accumulate more trehalose in the cell than the original strain. These data suggest that the ability to change its membrane fatty acid composition and to accumulate trehalose may make the isolated strain easily adapt to changes in external condition.

• Microbiology and Biotechnology Letters
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• v.22 no.6
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• pp.584-592
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• 1994

Several mutation procedures have been compared to obtain an ethanol-tolerant Saccha- romyces diastaticus strain secreting glucoamylase. These procedures include spontaneous mutation, EMS treatment, UV irradiation, and combination of EMS treatment and UV irradiation. All these methods were followed by adaptation of the yeast cells to gradually higher ethanol concentration. Among these procedures, the combined method of EMS treatment and UV irradiation gave the promising result, i.e. the ethanol tolerance of the yeast increased from 11.5%(v/v) to 14.0%(v/v). Respiratory deficient petite mutants of industrial and ethanol-tolerant yeast strains have been isola- ted and hybridized with haploid S. diastaticus strains. The resulting hybrids showed increased ethanol tolerance and starch-fermentability.

• Korean Journal of Weed Science
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• v.15 no.3
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• pp.214-223
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• 1995

Tolerant corn cultivars to bentazon were selected and tolerance mechanism of corn cultivars to bentazon was studied by determining bentazon 6-hydroxylase(B6H) activity which was known to detoxify bentazon to 6-hydroxy bentazon at induced enzyme conditions with treatments of 1,8-naphthalic anhydride, ethanol and phenobarbital. Tolerant cultivars to bentazon were selected by growth response of corn by foliar application of bentazon to corn cultivars. Kwanganok, GA 209, IK 2, DB 544, and Suwon 19 were tolerant to bentazon, but KSS 3, KSS 4, KS 5, and Danok 2 were susceptible. Pretreating corn seeds with 1,8-naphthalic anhydride increased B6H activity at all cultivars, but the tendencies were more remarkable at Suwon 19 and GA 209, tolerant cultivars, than at Danok 2 and KS 5, susceptible cultivars. Treating corn shoots with ethanol increased B6H activity at Suwon 19 and GA 209. B6H activity was enhanced by treatments of ethanol at 1.0 or 2.5%, but decreased at ethanol 2.5 or 5.0% at Danok 2 and KS 5. Treating corn shoots with phenobarbital increased B6H activity at Suwon 19, GA 209, Danok 2, and KS 5 by treatments of phenobarbital at 2.0mM, but decreased at 4.0 or 8.0mM at all cultivars. Therefore, the tolerant mechanism of corn cultivars to bentazon may be explained partially by the activity of bentazon 6-hydroxylase which detoxifies bentazon to 6-hydroxy bentazon.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.184-189
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• 2012

We sought to breed an industrially useful yeast strain, specifically an ethanol-tolerant yeast strain that would be optimal for ethanol production, using a novel breeding method, called genome reconstruction, based on chromosome splitting technology. To induce genome reconstruction, Saccharomyces cerevisiae strain SH6310, which contains 31 chromosomes including 12 artificial mini-chromosomes, was continuously cultivated in YPD medium containing 6% to 10% ethanol for 33 days. The 12 mini-chromosomes can be randomly or specifically lost because they do not contain any genes that are essential under high-level ethanol conditions. The strains selected by inducing genome reconstruction grew about ten times more than SH6310 in 8% ethanol. To determine the effect of mini-chromosome loss on the ethanol tolerance phenotype, PCR and Southern hybridization were performed to detect the remaining mini-chromosomes. These analyses revealed the loss of mini-chromosomes no. 11 and no. 12. Mini-chromosome no. 11 contains ten genes (YKL225W, PAU16, YKL223W, YKL222C, MCH2, FRE2, COS9, SRY1, JEN1, URA1) and no. 12 contains fifteen genes (YHL050C, YKL050W-A, YHL049C, YHL048C-A, COS8, YHLComega1, ARN2, YHL046W-A, PAU13, YHL045W, YHL044W, ECM34, YHL042W, YHL041W, ARN1). We assumed that the loss of these genes resulted in the ethanol-tolerant phenotype and expect that this genome reconstruction method will be a feasible new alternative for strain improvement.

• Applied Biological Chemistry
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• v.34 no.1
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• pp.67-74
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• 1991

In order to improve the productivity of ethanol by sugar-alcohol-tolerant Saccharomyces cerevisiae D1, the effect of addition of $Ca^{2+}$ on the alcohol fermentation was investigated. The addition of $Ca^{2+}$led to both the improvement of ethanol productivity and the increase of viable cell concentration. The optimal concentration of $Ca^{2+}$ was 0.8 mM. The higher was initial concentration of glucose, the greater effect of $Ca^{2+}$ was observed. Ethanol inhibition of growth, specific death rate in lethal concentration of ethanol, and extracellular final pH decreased by the addition of $Ca^{2+}$. The effect of $Ca^{2+}$ supplementation was explained by the increase of its ethanol tolerance.

• Microbiology and Biotechnology Letters
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• v.23 no.6
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• pp.723-729
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• 1995

A flocculating sugar tolerant yeast strain was isolated from fermenting Takju. This strain was identified as Saccharomyces cerevisiae CA-1 according to the Lodder's yeast taxonomic studies. The isolated yeast could grow in 50% glucose and in 7% ethanol in the YPD medium. It's optimal growth temperature, initial pH, shaking rate and initial glucose concentration for ethanol fermentation showed 35$\circ$C, 4.5, 150 rpm, 15%, respectively. Ethanol concentration was 63 g/l in 20% glucose after 24 hours, fermentation yield was 0.49 g-ethanol/g-glucose in 10% glucose after 24 hours and ethanol productivity was 3.09 g/l$\cdot $h in 10% glucose after 12 hours in batch fermentation. Repeated batch fermentation was possible for over 50 days and ethanol yield, ethanol productivity and substrate conversion rate were 0.39-0.50 g/g, 1.63-2.08 g/l$\cdot $h and more than 99%, respectively during these periods.

• Applied Biological Chemistry
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• v.34 no.1
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• pp.61-66
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• 1991

In order to improve the productivity of ethanol by sugar-alcohol-tolerant Saccharomyces cerevisiae D1, the effect of addition of soybean meal on the alcohol fermentation was investigated. The addition of soybean meal led tn the increase of the ethanol productivity and viable cell concentration. Increasing the mont of soybean meal increased the number of viable cells and the consumption percentage of glucose. The water-soluble fraction of soybean meal was nearly as effective as whole-soybean meal, whereas the lipidic fraction had no positive effect. The addition of 4% soybean meal increased the rate of ethanol production regardless of the initial concentrations of glucose. The rate of glucose consumption fermenting a soybean meal supplemented medium was higher than possible in a non-supplemented medium, either in the absence or in the presence of ethanol. But the percentage of ethanol inhibition of the glucose consumption rate was identical for supplemented md unsupplemented media. The increase of final ethanol concentration could not be attributed In an increase of ethanol tolerance of yeast cells but to the satisfaction of nutritional deficiencies.