• Korean Journal of Microbiology
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• v.30 no.3
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• pp.160-163
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• 1992

Killer toxin from killer yeast fusant FWKS 260 developed by protoplast fusion between the wild killer yeast and alcohol-fermenting yeast was purified by ammonium sulfate fractionation. Amicon PM I0 concentration. Sephadex G-200 and Scphadcx G-75 column chromatography. The purified killer toxin showed a single band by SIX-polyacvlamide gel electrophoresis. The protein part of killer toxin was active site. which was found by treating the proteolytic enzyme such as pronase E and pepsin to killer toxin. The killer toxin was stable at pH 2.0-5.0 and 20$^{\circ}$C. but inactivated with increasing temperature. The molecular weight was determined to be approximately 13.000 according to the results obtained from the SDS-polyacrylamide gel electrophoresis. It was confirmed that the purified killer toxin is glycoprotein by showing a red single band after st'tining with Schiffs reagent.

• KSBB Journal
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• v.14 no.4
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• pp.434-439
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• 1999

A wild yeast S-13 which has excellent killer toxin activity to gas-producing yeast of traditional Doenjang and Kochujang was selected among forty seven strains of Meju yeasts and identified as Hansenular capsulata S-13 by investigation of the morphological, cultural and physiological properties. The optimal conditions for the production of killer toxin were investigated. H. capsulata s-13 showed the higest killer toxin activities when it was cultured up to the late-log phase of 36 hr in YEPD medium (pH4.5) at $25^{\circ}C$ H. capsultara S-13 showed killer toxin activities to seven strains of industrial yeasts such as S. cerevisiae, C. veratilis and P. membranaefacieus.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.8
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• pp.1077-1082
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• 2007

In this study, killer yeasts were isolated from traditional Nuruk to improve storage and suppress contaminant in food industry. Among killer yeasts, yeast K15 showed strong killer toxin activity and inhibited growth of Salmonella Typhimurium and Vibrio parahaemolyticus. Killer yeast K15 was identified with Pichia anomala by the Microlog TM 4.0 identification system and homology of the ITS sequence. Killer toxin generated from P. anomala K15 was inactivated by pronase E and suggested to be a protein. Therefore killer toxin of P. anomala K15 was thought to be safe in human such as bacteriocin. P. anomala K15 was sufficient for growth in 50% glucose and could be used to prevent contaminant in initial stages of alcohol beverage fermentation.

• The Korean Journal of Food And Nutrition
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• v.13 no.2
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• pp.158-163
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• 2000

Killer yeast, Hansenular capsulata S-13 were treated with heat, ethylmethane sulfonate and N-methyl-n'-nitro-n-nitrosoguanidine and a mutant(S13-E1), showing 2-fold higher killer toxin activity than that of parent strain to killer sensitive strain, Saccharomyces cerevisiae ATCC 38026 was obtained. Hansenular capsulata S13-E1 showed strong killer toxin activity to Saccharmyces mellis and Saccharomyces sal년 and four strains of gas-producing yeasts from traditional Doenjang and Kochujang. The culture condition for killer toxin production by Hansenular capsulata S13-E1 was optimized to be 1.0% potato extract, each 0.5% of peptone and glucose, and 0.025% MgSO4 with initial pH 4.5 at 3$0^{\circ}C$ and 36 hr of batch cultivation.

• Korean Journal of Microbiology
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• v.27 no.4
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• pp.415-421
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• 1989

Ten strains out of about 1,000 yeast strains isolated from byproducts of alcoholic industries, milk products, fruits, greens, food-related industries and soils of nature, revealed the killer activities. Two strains which have excellent killer activities among them were isolated and identified with Saccharomyces cerevisiae B 15-1 and Hansenula anomala Y 33 by investigation of the morphological, cultural and physiological properties. The optimal conditions on these strains for the production of killer toxin were investigated. The strain B 15-1 showed the highest killer toxin activities when it was cultured up to the log phase of 48 hr in YPD medium (pH 4.7) at $25^{\circ}C$. On the other hand, the strain Y33 revealed the highest activities when it was cultured up to the stationary phase of 60 hr in YPD medium (pH 4.0) at $20^{\circ}C$. The sensitive strain Kyokai 7 was found to be killed entirely by the killer toxin produced from the wild killer yeast B 15-1 when B 15-1 was cocultured with the same cell concentration ($10^{6}$ cells/ml) of Kyokai 7 after cultivation of 36 hr, and with large concentration ($9\times 10^{7}$ cells/ml) after 48 hr.

• Korean Journal of Microbiology
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• v.30 no.3
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• pp.177-180
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• 1992

Among 120 U. maydis strains isolated in Korea 14 different strains containing specific viral dsRNA segments were analyzed for the distribution of dsRNA and the production of toxin protein. Several distinctive dsRNA patterns were identified, 9 cases of P type with typical H, M and L ds RNA and one case of non-P-type, the frequency of a specific isolate was decreased with increasing number of dsRNA segments. The presence of dsRNA had no effect on the cultural or morphological phenotype of the host. Two isolates containing P type dsRNA segments appeared to produce toxin protein (killer strains) which inhibited the growth of 4 isolates (sensitive strain) with different susceptibility. Two killer strains contain unique M dsRNA segment which may code for toxin protein. However, the presence of toxin-sensitive strains among dsRNA-free isolates was similar to that of ds RNA containing strains.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.26-30
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• 1990

Candida dattila K109 and K112 isolated from grapes in Korea showed killer activity toward Saccharomyces cereuisiae 5 x 47, S. cereuisiae 1368, Hamenula, Torulopsis, Kluyueromyces, Debaryomyces, and Brettanomyces, and showed the most effective killer activity at 22-26$^{\circ}C$ and at pH 3.9-4.1. The killer actvity of both toxins were remarkably decreased at higher temperature than $25^{\circ}C$ and higher pH than pH 4.0. And the toxins were suggested to be glycoproteins inactivated by pronase E and pepsin. The killer activity was not cured by incubation at elevated temperature of 30-37"C, but cured by treatment with 0.0105-0.3 ppm cycloheximie.imie.

• Journal of Microbiology and Biotechnology
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• v.10 no.4
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• pp.547-550
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• 2000

The yeast Williopsis californica was shown to secrete a unique broad-spectrum killer toxin (Wicaltin) with antifungal activity against 14 yeast genera, including yeast-like and mycelial forms of the human pathogens Candida albicans and Sporothrix schenkii. Agar diffusion bioassays indicated that its activity was more pronounced than the antifungal potential of frequently used antimycotics; 0.07 pmol Wicaltin showed the same toxicity as 0.2 pmol miconazole and 29 pmol clotrimazole. Since the toxin's primary target would appear to be the yeast cell wall, Wicaltin may be attractive in combatting clinically relevant yeast and fungal infections.

• BMB Reports
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• v.31 no.2
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• pp.123-129
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• 1998

Foreign proteins, $endo-{\beta}-1,4-glucanase$ of Bacillus subtilis, preS1+S2 region of hepatitis B virus large surface antigen, human ${\beta}_2-adrenergic$ receptor ($h{\beta}_{2}AR$), and bovine growth hormone (bGH) were expressed in Saccharomyces cerevisiae and secreted into the medium. These proteins were expressed using the alcohol dehydrogenase I (ADH1) promoter of Saccharomyces cerevisiae and secreted by signal sequence of the 97 K killer toxin gene of doublestranded linear DNA plasmid (pGKL1) of S. cerevisiae. All these proteins underwent severe modifications; in particular, N-glycosylation in the case of $endo-{\beta}-1,4-glucanase$, $h{\beta}_2AR$, and preS1+S2. Seventy four percent of the expressed $endo-{\beta}-1,4-glucanase$ was secreted into the culture medium. Highly modified proteins were detected in the culture medium and in the cell. Expressed $h{\beta}_2AR$, which has seven transmembrane domains, remained in the cell. The degrees of secretion and modification and the states of proteins in the culture medium and in the cell were quite different. These results indicated that the nature of the protein has a critical role in its secretion and modifications.

• Journal of Microbiology
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• v.35 no.4
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• pp.323-326
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• 1997

Toxin protein from Ustilago maydis virus SH14 isolated in Korea was purified using ethanol precipitation, cation exchange, gel filtration and anion exchange chromatography. The molecular weight of the purified protein was estimated to be 8.3 kDa by SDS-PAGE analysis. The Nterminal sequence of the protein is L-G-I-N-C(K)-R-G-S-S-Q--C(K)-G-L-S-G which is highly homologous with that of P4 toxin, but the amino acid composition and electrophoretic mobility in a native PAGE of the toxin protein were totally different from those of P4 toxin respectively. The SH14 toxin was shown to have immunological cross-reactivity about 50% with P4 toxin when examined by Western hybridization.