• Food Science and Biotechnology
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• v.14 no.6
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• pp.878-880
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• 2005

Antimutagenic effects of methanol extracts of various soybean pastes against aflatoxin B1 were examined using Salmonella typhimurium TA98 and TA100. Antimutagenic activities of boiled soybeans, Japanese Miso, traditional Korean soybean pastes, soybean pastes fermented by wild type strains, and soybean pastes fermented by mutants, transformants, and cell fusants were 53.6 to 54.6%, 73 to 79.7%, 78.3 to 95.7%, 85 to 97.1%, 71.9 to 78.3%, 65.5 to 77.7%, and 73.4 to 79.0%, respectively. Soybean pastes fermented by wild type strains showed higher activities than those fermented by mutant, transformant, and cell fusant strains.

• Journal of Microbiology and Biotechnology
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• v.3 no.1
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• pp.24-30
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• 1993

In order to develop a novel yeast which produces the charateristic aroma of soy sauce, a protoplast fusion between Zygosaccharomyces rouxii WFS4 and Torulopsis versatilis IAM 4993 was carried out. Auxotrophic mutants as selective markers were obtained from Zygosaccharomyces rouxii and Torulopsis versatilis by treatment of N-methyl-N -nitro-N-nitrosoguanidine. The conditions of the protoplast formation and the regeneration for fusion were examined. The protoplast fusion using polyethylene glycol 4000 led to the fusion frequency of $4~5{\times}10^{-7}\;cells/ml$. Among fusants, a fusant ST723-F31 presented the best results in terms of the aromaticity of fragrance, the growth pattern, the resistance against salt and the degree of growth according to pH. It makes easy to control the production and the balance of aroma components so that it gives a good flavor, shortens the fermentation period and, simplifies the preparation process when using a bioreactor into which fusant is immobilized.

• Microbiology and Biotechnology Letters
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• v.36 no.2
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• pp.158-164
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• 2008

The fusants which contain starch utilizing ability and alcohol fermenting ability were developed by protoplast fusion of Saccharomyces cerevisiae KOY-1 and Saccharomyces diastaticus KCTC 1804. Sacharomyces cerevisiae KH-12 was obtained by haploid induction from Saccharomyces cerevisiae KOY-1. The auxotropic mutants of yeast were obtained by using an ethylmethane sulfonate (EMS). The frequency of protoplast formation in Saccharomyces cerevisiae KOY-1 $(Met^-)$ and Saccharomyces diastaticus KCTC 1804 $(Trp^-)$ were 90.5% and 97.7%, respectively. The frequency of fusant formation was $1.79{\times}10^{-4 }$ for the regenerated protoplast and the 1,000 fusants were obtained. Fusant FA 776 was selected as a potential yeast which contain an alcohol fermenting ability in the starch medium. The genetic stability was 4.64% for 10 passages of generation. Fusant FA 776 produced 13mg/ml of alcohol in 24% starch medium and showed 1.86-fold higher alcohol fermenting ability than Saccharomyces diastaticus KCTC 1804.

• The Korean Journal of Mycology
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• v.17 no.3
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• pp.137-144
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• 1989

Effects of incompatibility existing between intra-and interspecies in Pleurotus spp. on protoplast fusion, clamp formation of their fusants and fruitbody production were investigated. Protoplast fusion between intra-and interspecies of the fungus was achieved by Poly ethylene glycol treatment. The fusion frequency between intraspecies was a little higher than that of interspecies. Fusion frequency between interspecies was not correlated with their similarities based on isozyme patterns. In case of protoplast fusion between intra-and interspecies, the fusants from the compatible isolates produced normal fruit bodies, while those from the incompatible isolates did not produce clamp connections and fruit bodies except those of a few isolates presumed mutants.

• The Korean Journal of Mycology
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• v.17 no.3
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• pp.119-123
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• 1989

Interorder heterokaryons were obtained by polyethylene glycol induced fusion of protoplasts from auxotrophic mutants of Pleurotus ostreatus in agaricales and Ganoderma applanaturm in aphyllophorales. When transferred to MMM plates, all fusion colonies exhibited an extremely growth rate. During three times subcultivation on MCM the growth rate of fusants showed faster little by little. Seventy-five % fusion products of potoplasts showed mixed morphologies between those of P. ostreatus and G. applannatum in the first subcultivation on MCM and MGM. The phenotype of these fusants changed similar those of P. osteatus type after three times subcultivation on MCM. However, phenotype of 25% stable strains did not change on subcultivation. Hyphae of all fusion products did not form true clamp connection. All these types did not produce primordia. A comparrison of interorder somatic hybrids was made using isozyme analysis of esterase, malate dehydrogenase and peroxidase. In most cases the enzyme patterns of G. applanatum were not distinct, however, fusant showed non-parental bands.

• The Korean Journal of Mycology
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• v.22 no.4
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• pp.378-385
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• 1994

Interspecific somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus and Pleurotus sajor-caju. The fusion products between incompatible strains did not form clamp connections. Fruiting body of the clampless fusants was induced by light-dark cycle on saw-dust-rice bran substrate in glass bottles. Out of them, seven somatic hybrids produced fruiting bodies of intermediate morphology of the two species. Light and low temperature were the initiating factors for the development of clamped hyphae from the clampless mycelial colonies. All of these basidiocarps had clamp connections. Eight fusants from the six crosses were analysed with the segregation of genetic characters by random spore isolates. In the three combinations, unexpected alleles were shown. Somatic hybrid between P188 (P. ostreatus 2-1 + P. sajor-caju 2-53) and P. florida 2-3 by triple cross produced fruiting bodies similar to those of fusant between P. ostreatus and P. florida. All the genetic charaters from the three strains were shown to segregate and recombine.

• The Korean Journal of Mycology
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• v.16 no.2
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• pp.79-86
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• 1988

Interspecific fusion products were obtained from fusion of Ganoderma applanatum and Ganoderma luridum. Frequency of fusion was 0.77-1.38%. Fusion products were selected by the comparision of morphology and color of colony. Fusion Products had chracteristics of two parental strains and generally grew faster than the parents. Some fusants were segregated on GCM. Fusion products were confirmed by mycelial morphology and electrophoretics pattern of esterase isozyme from mycelium. Most of fusion products were lack in clamp connection but those fusion products, that were segregated produced mycelium with and without clamp connection. Some of the fusion products produced fruit body on sawdust medium.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.611-617
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• 1990

This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.228-232
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• 2003

Lovastatin produced by Aspergillus terreus via polyketide pathway is a secondary metabolite with high anti-hypercholesterolemic activity. In this paper we are going to present effective strain development strategies for lovastatin production by comparing the productivity of the mutants obtained through traditional rational screening process and protoplast fusion method. Mutants resistant against various antibiotics and/or antimetabolites showed significantly higher lovastatin productivity than the corresponding mother strains, demonstrating that rational screening method was very efficient in selecting high yielding producers. Recombinant fusants obtained using protoplast fusion between high producers were observed to have very different morphology and physiology as represented by the production and secretion of lovastatin, as well as cell growth pattern. In parallel with the strain development, optimization process for the production medium was carried out in order to find optimal concentrations of the medium components using such a powerful statistical method as response surface method (RSM). It was concluded that not only the optimum production medium but also good morphological characteristics of the high-yielding producers led to higher lovastatin production.

• Microbiology and Biotechnology Letters
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• v.13 no.4
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• pp.383-389
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• 1985

Optimum conditions of PEG treatment for the intergeneric fusion of yeast protoplasts were investigated. Fusants were selected by nutritional complementation on minimal medium. The intergeneric fusion frequency between pro-toplasts of S. cerevisiae and C. tropicalis was distributed 10$^{-4}$ to 10$^{-6}$, depending on the combination of parental strains. PEG 4000 or 6000 are equally effective. 30%(w/v) PEG 4000 was found to be optimum and below 20% its stabilizing effect was lost, resulting in protoplast lysis, and optimum pH was 8.0. The efficiency of PEG was enhanced by higher temperature of the PEG solution, and by the addition of Ca ions. The stimulating effect of Ca ions in the range of 1 mM to 100 mM proved similar.