• Title/Summary/Keyword: Protoplast fusant

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The Interspecific Protoplast Fusion between S. peucetius subsp. caesius and S. platensis (S. peucetius subsp. caesius와 S. platensis의 원형질체 융합 및 융합균주의 분리)

  • Im, Mi-Song;Lee, Kang-Man
    • YAKHAK HOEJI
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    • v.38 no.6
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    • pp.696-702
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    • 1994
  • An interspecific fusant strain, MS1, was obtained by protoplast fusion between S. peucetius subsp. caesius and S. platensis. We studied on the microbiological and cultural characteristics of the fusant MS1. In liquid culture, the viscosity of culture broth of S. peucetius increased during incubation However the fusant MS1 formed pellet like S. platensis without viscosity change. On agar medium the colony morphology of MSI resembled S. platensis but the color was similar to S. peucetius. The fermentation products of the fusant MS1 was identical with S. peucetius.

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A study on strain improvement by protoplast fusion between amylase secreting yeast and alcohol fermenting yeast - III. Isolation and characterization of fusant between S. diastaticus and C. tropicalis (Amylase분비효모와 alcohol발효효모의 세포융합에 의한 균주의 개발 - 제3보. S. diastaticus와 C. tropicalis 간의 세포융합 및 융합체의 성질-)

  • 서정훈;권택규;홍순덕
    • Microbiology and Biotechnology Letters
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    • v.14 no.5
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    • pp.359-363
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    • 1986
  • S. diastaticus hydrolysised $\alpha$-1.4 linkage of the starch and was known fermenting yeast strain, but poorly hydrolized $\alpha$-1.6 linkage of the starch. To improve the starch fermentation ability of yeast, we tried that protoplast fusion between S. diastaticus and C. tropicalis and finally two starins of fusant (FPDC42, FPDC43) were obtained. C. tropicalis well hydrolysis both $\alpha$-1.4 and $\alpha$-1.6 linkanges in the starch. The protoplast of parental auxotrophic cells were fused in the presence of 10mM CaCl$_2$ and 35% of polyethylene glycol (M. W. 4,000). The fusion frequency was 10$^{-5}$ to 10$^{-6}$. Properties of the fusants(genetic stability, assimilation of carbon sources, random spore formation, copper resistance, NaCl tolerance, DNA content, cell size and growth rate) were investigated.

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Interspecific Protoplast Fusion between Fusarium poae and Fusarium sporotrichioides (Fusarium poae와 Fusarium sporotrichioides간의 원형질체 융합)

  • 하경란;장성렬;민병례
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.123-129
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    • 1991
  • In order to develop the protoplast fusion method of the strains of Fusarium, the interspecific protoplast fusion was attempted between Fusarium poae and F. sporotrichioides. Various auxotrophic mutants were isolated by the treatment of N-Methyl-N'-Nitro-N-Nitrosoguanidine. The optimal conditions for the formation and regeneration of protoplasts were examined and the characteristics of a fusant were studied. As a results, protoplasts were readily obtained from 18 hours cultured mycelia by the treatment of driselase for 3 hours and 0.6 M KCl as a best osmotic stabilizer at pH 6.0 for the formation of protoplast. Sucrose was the most suitable for the regeneration. Polyetylene glycol (M.W. 8,000) in $CaCl_{2}$-glycine solution was used to induce the protoplast fusion. The interspecific fusion frequency between protoplasts among the auxotrophic mutants of the two strains ranged from $2.7*10^{-2}$ to $5.7*10^{-3}$ . DNA content and cellulase activity were rather increased in the interspecific fusant. The lag phase of growth curve was slightly elongated in the fusant.

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Development of a Novel Yeast Strain Which Ferments Soy Sauce by Protoplast Fusion

  • Lee, Eun-Ju;Kim, Jong-Kyu
    • 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.

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Characterization of Alcohol Fermentation and Segregation of Protoplast Fusant of Saccharomyces cerevisiae and Pichia stipitis

  • YOON, GEE-SUN;TAE-SIK LEE;CHUL KIM;JIN-HO SEO;YEON-WOO RYU
    • Journal of Microbiology and Biotechnology
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    • v.6 no.4
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    • pp.286-291
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    • 1996
  • A study was conducted to investigate the characteristics of segregation and alcohol fermentation of intergeneric fusants. The protoplast fusion of both Pichia stipitis CBS 5776 and Saccharomycess cerevisiae STV 89 was carried out. The fusion frequency was $5\times10^{-8}$ and among fusants selected, a fusant F5 showed the best results in ethanol production by sucrose and xylose fermentations. The performance of xylose fermentation by this fusant was better than that of P. stipitis CBS 5776 and fusant F5 exhibited sucrose fermentation patterns intermediate to the two parent strains. The fusant F5 was segregated into a pair of parental strains during the several culture passages. In the average, 91$%$ of colonies had a similar characteristics of P. stipitis while 7$%$ of colonies resembled S. cerevisiae. Only 2$%$ of colonies had the characteristics of the original fusants. At the sixth passage, all segregants resembled P. stipitis. From these results it is suggested that intergeneric protoplast fusion led to an integration of S. cerevisiae genes, rather than whole chromosomes, within the entire genome of P. stipitis.

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The Fermentation Characteristics of the Interspecific Protoplast Fusant of S. peucetius and S. platensis (Streptomyces peucetius subsp. caesius와 S. platensis사이의 원형질체 융합균주의 발효특성)

  • Im, Mi-Song;Lee, Kang-Man
    • YAKHAK HOEJI
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    • v.38 no.6
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    • pp.749-755
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    • 1994
  • An interspecific fusant strain, Streptomyces MS1 was obtained by protoplast fusion between S. peucetius subsp. caesius and S. platensis. We studied on the fermentation characteristics of the fusant strain. The fermentation products of the fusant MS1 was identical with S. peucetius, but its production of anthracycline was more stable than S. peucetius under various fermentation conditions in regard to acidogenesis of fermentation broth. The optimal medium composition for anthracycline production by fusant MS1 as follows: sucrose 2.0%, glucose 1.0%, soytone 0.7%, $CaCO_3$ 0.2%, $KH_2PO_4$ 0.013%, casamino acids 0.01%, $K_2SO_4$ 0.025%, $MaCl_2\;6H_2O$ 1.024%, 5M $CaCl_2\;5H_2O$ 0.4%, 1N NaOH 0.7%, 20% L-proline 1.5%. In this condition, the productivity of anthracycline was $80{\sim}100\;{\mu}g/ml$.

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Enhancement of Ethanol Tolerance of Lactose Assimilating Yeast Strain by Protoplast Fusion

  • Ryu, Yeon-Woo;Jang, Heang-Wook;Lee, Haing-Sook
    • Journal of Microbiology and Biotechnology
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    • v.1 no.3
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    • pp.151-156
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    • 1991
  • In order to construct a yeast strain having high ethanol tolerance together with good lactose fermentation ability, the protoplast fusion using Saccharomyces cerevisiae STV 89 and Kluyveromyces fragilis CBS 397 was carried out. Auxotrophic mutants of K. fragilis were obtained as a selection marker by treatment of ethylmethane sulfonate. The best mutant for protoplast fusion was selected based on the capabilities of ${\beta}-galactosidase$ production and lactose fermentation. The protoplast fusion using polyethylene glycol and calcium chloride solution led to the fusion frequence of $3{\times}10^{-6}$ and a number of fusants were obtained. Among these fusants, a fusant F-3-19 showed the best results in terms of ethanol tolerance, ${\beta}-galactosidase$ activity and lactose fermentation. The performance of lactose fermentation and ethanol tolerance by this fusant were better than those of K. fragilis. Study on the ethanol tolerance having relation to fatty acid composition and intracellular ethanol concentration revealed that the fusant F-3-19 had a higher unsaturated fatty acids content and accumulated less amount of intracellular ethanol compared with a parent of K. fragilis.

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Antitumor Activity of the Intergeneric Protoplast Fusant between Lentinus edodes and Coriolus versicolor (표고와 운치의 원형질체 융합균주의 항암작용)

  • 곽은경;김하원;심미자;현진원;김병각
    • Biomolecules & Therapeutics
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    • v.8 no.3
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    • pp.235-240
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    • 2000
  • Antitumor effect of LC43, a protein-bound ploysaccharide (M.W. 43 kDa) that was purified from intergeneric protoplast fusant of Lentinus edodes and Coriolus versicolor, was elucidated against mouse sarcoma 180 cell in vitro and in vivo. By injecting LC43 into ICR mice bearing solid or ascitic sarcoma 180, tumor regression and survival rates were investigated. To examine the effects of LC43 on immunopotentiation activity. immunoorgan weight, B cell differentiation, T cell activity and macrophage activation were determined. LC43 showed antitumor effects against both solid tumor and ascitic tumor of sarcoma 180. It did not change significantly the immunoorgan weight but potentiated immune responses such as B cell differentiation and the release of superoxide anion from macrophages. These results suggest that the protein-bound polysaccharide of LC43 exhibited antitumor activities through the activation of immune-related cells and acted as an immunmodulator.

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Characterization of fusants between thermophilic clostridium thermocellum and clostridium thermohydrosulfuricum (고온성 clostridium thermocellum과 clostridium thermohydrosulfuricum의 융합체의 특성)

  • 김욱한;정기택;이용현
    • Korean Journal of Microbiology
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    • v.28 no.4
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    • pp.311-317
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    • 1990
  • Intraspecific and interspecific protoplast fusions in/between C. thermocellum and C. thermohydrosulfuricum were studied. Protoplast fusions were well induced in 30-40% PEG solution, however, their fusion frequencies were low level of 1.2*10$^{-7}$ for intraspecific fusion of C. thermocellum, $6.7*10^{-7}$ for C. thermohydrosulfuricum, and 4.2*10$^{-7}$ for interspecific fusion between above two Clostridia, respectively. Most fusants were unstable and segregated after 3 subcultures. Relatively stable intraspecific C. thermocellum fusant FTT17, intraspecific C. thermohydrosulfurecum fusant FSS22 and interspecific fusant FTS3, which were stable after several subcultures, were selected and properties of fusants were further investigated, Phenotypes of the fusants were similar with wild types mostly in cellular morphology, carbon source assimilation and enzyme activities. However they were differed in assimilation of pyruvic acid and sorbitol as carbon source. The DNA contents of fusants were slightly increased compared with wild types. Ethanol production by intraspecific and/or interspecific fusants was not increased, however, acetic acid production as byproduct was decreased or not detected, which indicates that industrial thermophilic anaerobes can be improved by means of protoplast fusion of two strains.

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Functional Properties of a Interspecific Protoplast Fusant from Lactobacillus bulgaricus and Lactobacillus helveticus (유산균의 이종간(異種間) 세포융합균주의 기능특성)

  • Jun, Hong-Ki;Park, Hyun-Jeong;Song, Jae-Chul;Baik, Hung-Suk
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.310-315
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    • 1993
  • An interspecific fusant was made from the protoplasts of two strains of Lactobacillus genus (L. bulgaricus and L.helveticus). And in order to test the applicability of the fusant in manufacturing the cheddar cheese, functional properties of the strain was examined by determining acid-producing activity, three important enzyme activities and volatile free fatty acid-producing activity. The recombinant strain did not exhibit greatly increased acid-porducing activity. Lipase and volatile free fatty acid-porducing abilities of the fusant, however, were remarkably higher than those of the two parental strains. The fusant actually porduced the cheese porduct of the highest ammount of total volatile free fatty acid after 7 days ripening at 10$^{\circ}C$. Finally, the cheddar cheese ripened with this strain was also evaluated to be high preference and flavor intensity by organoleptic panel tests.

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