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effect of Nif A Overproduction on the Improvement of Nitrogenase Activity of Klebsiella oxytoca Strains (Nif A의 Overproduction에 의한 Klebsiella oxytoca 균주의 질소 고정력 증진 효과)

  • 서현창;유익동
    • The Korean Journal of Food And Nutrition
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    • v.5 no.2
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    • pp.137-143
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    • 1992
  • As a research for developing biofertilizers, Klebsiella oxytoca, an associative nitrogen fixer in the rhizosphere of rice plant in the soil of paddy field, was subjected to molecular breeding. The results obtained were as followings. 1). By transforming pbIC71A, Nif A overproducing plasmid, into Klebsiella oxytoca NGl3, Klebsiell6f oxytoca SH3l, and Klebsiella oxytoca SH161, nitrogenase activities in the absence of nitrogen source in the medium were increased 6.4, 17.2, and 13.5 times, respectively, in comparison with the parent strains. 2). Nitrogenase activity of Klebgiella oxytoca NGl3, Klebsiella oxytoca SH3l, and Klebsiella oxytoca SH161 was completely repressed In the presence of 15mM NH4+. But, nitrogenase activities of Klebsiella oxytoca NGl3/PMC71A, Klebsiella oxytoca SH3l /PMC71A, and Klebsiella oxytoca SH 161/pMC714 harboring PMC71A, were 13.7%, 7.7%, and 6.2% of the nitrogenase activities in the absence of nitrogen source in the medium, respectively.

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Studies on the Citric Acid Fermentation by Fungi (Part II) Preservation of the Selected Strains and the Effect of UV-Irradiation (사상균에 의한 구연산효소에 관한 연구 (제II보) 선정균의 보존 및 자외선조사 효과)

  • 성낙계;김명찬;심기환;정덕화
    • Microbiology and Biotechnology Letters
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    • v.8 no.2
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    • pp.87-92
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    • 1980
  • These experiments were conducted to manage more safe the selected strains and improve their characteristics. The results obtained were as follows: Preservation on soil at the range of 0 to 5 C was suitable and there were no remarkable changes on their abilities to produce citric acid until 10 months preservation. The successive transfer of spore slightly stimulated the selected strain, M-315 to produce citric acid and the spore precultured on sporulation medium for 7-10 days was desirable as inoculum. Under UV-irradiation from the selected strains, 109 mutants whose morphological characteristics were changed were isolated. Among them, the mutant M-80-12 was shown 3.2% increase on acid producing ability than that of its parent.

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Protoplast fusion of Aspergillus oryzae (Aspergilluis oryzae의 원형질체 융합)

  • 이수연;이주실;이영록
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.216-220
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    • 1989
  • As the bsic study about protoplast fusion of amylolytic fungus Aspergillus oryze and nonamyloytic sugar fermenter, Saccaromyces cerevsisae, the intraspecific protoplast fusion of A. oryzae was carried out and the properties of the obtained fusants were investigated. For protoplast fomation from mycellia of auxotrophs, Novozyme 234 as lytic enzyme was the most effective and optimal pH was determined to be pH 5.5-6.0. When the two types of protoplasts were treated with a fusogen including 30% PEG4000, they fused effectively and most of fusants were heterokaryons. Protoplasts aggregated with 30% PEG4000 after fusion treatment were observed by the microscope. Protoplast regeneration frequency was 1.46 to 13.8% and complementation frequency of fusion was 0.12 to 0.16. Fusant strains had a 1.5-fold DNA content compared to that of parent strain. And amylase activity was intermediate between those of parent strains.

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Genotypic Identification in Commercial Strains of Pleurotus ostreatus based on AFLP and VCGs (AFLP 분석 및 체세포 불화합성에 의한 느타리 유사품종의 확인)

  • Seo, Kyoung-In;Yoo, Young-Bok;Jang, Kab-Yeul;Shin, Pyung-Gyun;Oh, YounLee;Kim, Kwang-Ho;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.14-20
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    • 2013
  • We already reported four groups which contains some similar strains based on URP-PCR in the previous paper. The objective of this study was to confirm those strains by the amplified fragment length polymorphism (AFLP) and vegetative compatibility group (VCG). AFLP analysis showed no difference among these strains except ASI 2595 and 2183 in Weonhyeong group and ASI 2829 in Suhan group. They showed specific DNA bands only in the result of P + AG/M + AAG and P + GT/M + ATG primer combinations out of eight different combinations. The AFLP primers produced a total of 330 fragments between 80 and 1000 bp in length for 31 Pleurotus ostreatus strains. At a genetic similarity of 0.96, the UPGMA analysis separated the isolates into four distinct clusters. Each group was classified by similar strains. Confrontation test by vegetative compatibility groups (VCGs) also showed distinct line between strains from different groups, but no line between similar strains within the cluster. Our results indicate that most of similar strains was not distinctness. Thus, similar strains are considered to be very close on the genealogy of their parent or same strain with different name.

Enzyme Production by the Mutant of Aspergillus oryzae (국균변이주(麴菌變異株)에 의한 효소생산(酵素生産))

  • Park, Joong;Sohn, Cheon Bae
    • Korean Journal of Agricultural Science
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    • v.13 no.2
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    • pp.279-288
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    • 1986
  • One hundred and fifty one mutant strains were obtained from the parent strain Aspergillus oryzae MF by ultra-violet ray irradiation. Among those mutants a strain, Asp. oryzae UM-36 which hyperprodued protease, was selected and its morphological characteristics and the production of enzymes protease, ${\alpha}$-amylase, and glucoamylase on wheat bran koji and on soy-sauce koji were studied. The results obtained were as follows 1. The selected mutant showed slower growth and weak sporulation on malt agar and on Czapek agar than the parent strain. 2. The conidiophores of the mutant were generally shorter than those of the parent when grown on malt agar. 3. Sectoring in colonies was not found when grown on malt agar and on Czapek agar. 4. The level of protease production by the mutant was increased approximately 1,4-fold higher on wheat bran koji and 2-fold higher on soysauce koji than by the parent. 5. The production of ${\alpha}$-amylase and glucoamylase by the mutant were also increased as compared with the parent on wheat bran koji and on soy sauce koji. 6. In the case of parent strain and mutant strain, the highest activity of protease appeared after three days in wheat bran medium at $30^{\circ}C$ incubation, but the highest activities of ${\alpha}$-amylase and glucoamylase appeared after two days.

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A study on strain improvement by protoplast fusion between amylase secreting yeast and alcohol fermenting yeast I. Isolation and characterization of fusant between S. cecevisiae and S. diastaticus (Amylase 분비효모와 alcohol 발효효모의 세포융합에 의한 균주의 개발 제1보. S. cerevisiae와 S. diastaticus간의 세포융합 및 융합체의 성질)

  • 서정훈;김영호;전도연;이종태
    • Microbiology and Biotechnology Letters
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    • v.14 no.4
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    • pp.305-310
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    • 1986
  • To improve the starch fermentation ability of yeast, hybrids were introduced by protoplast fusion of S. cerevisiae and S. diastaticus. The protoplasts of parental auxotrophic cells were fused in the presence of 10 mM CaCl$_2$and 30% of polyethyleneglycol (M.W 4, 000). The frequencies of fusant formation varied depending upon the strains used and were 3.51$\times$10$^{-4}$ to 5.04$\times$10$^{-4}$ for the regenerated protoplasts. The strains capable of extensive starch hydrolysis produce only 10% to total fusants. The 4 strains were finally selected by the results of starch fermentation and genetic stability test. The DNA content and cell volume of the fusants were greater than those of the parental strains.

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$^{31}p$ Nuclear Magnetic Resonance Studies of Acetic Acid Inhibition of Ethanol Production by Strains of Zymomonas mobilis

  • Kim, In-Seop;Barrow, Kevin D.;Rogers, Peter L.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.90-98
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    • 2003
  • In vivo $^31p$ Nuclear Magnetic Resonance ($^31p$NMR) and metabolic studies were carried out on an acetic acid tolerant mutant, Zymomonas mobilis $ZM4/Ac^R$, and compared to those of the parent strain, Z. mobilis ZM4, to evaluate possible mechanisms of acetic acid resistance. This investigation was initiated to determine whether or not the mutant strain might be used as a suitable recombinant host far ethanol production from lignocellulose hydrolysates containing various inhibitory compounds. $ZM4/Ac^R$ showed multiple resistance to other lignocellulosic toxic compounds such as syringaldehyde, furfural, hydroxymethyl furfural, vanillin, and vanillic acid. The mutant strain was resistant to higher concentrations of ethanol or lower pH in the presence of sodium acetate, compared to ZM4 which showed more additive inhibition. in vivo $^31p$ NMR studies revealed that intracellular acidification and de-energization were two mechanisms by which acetic acid exerted its inhibitory effect. For $ZM4/Ac^R$, the internal pH and the energy status were less affected by sodium acetate compared to the parent strain. This resistance to pH change and de-energization caused by acetic acid is a possible explanation for the development of resistance by this strain.

Selection of a L-Lysine-Overproducing Strain of the Red Seaweed Porphyra suborbiculata (Rhodophyta) through Mutation and Analog Enrichment

  • Luyen, Quoc-Hai;Chowdhury, Muhammad Tanvir Hossain;Choi, Jae-Suk;Kang, Ji-Young;Park, Nam-Gyu;Hong, Yong-Ki
    • Fisheries and Aquatic Sciences
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    • v.15 no.2
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    • pp.145-150
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    • 2012
  • An improved strain of the red seaweed Porphyra suborbiculata containing an increased amount of the essential amino acid L-lysine was obtained through mutation and analog enrichment. Mutagenesis using a 10% lethal dose of ultraviolet irradiation and an enrichment culture with the L-lysine analog aminoethyl-L-cysteine (AEC) was repeated to select the most productive strain using monospores of P. suborbiculata. The concentrations of AEC required to produce 50 and 100% inhibition of survival were 60 and 115 mM in the parent strain, and 72 and 135 mM in the selected AEC-resistant strain, respectively. The AEC-resistant strain, L130, produced 1.74-fold more lysine compared to its parent strain. Thus, mutagenesis with analog enrichment shows promise for selecting seaweed strains that can overproduce this essential amino acid.

Selection of High Tobramycin-Producing Mutants (Tobramycin 고생산성 변이주의 분리)

  • 나규흠;김학주;김기태;양중익;김계원
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.343-347
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    • 1991
  • An improved method for the selection of high tobramycin-producing mutants of Streptomyces tenebrarius ATCC 17920 was investigated. By the use of apramycin-containing media, low nebramycin-producing mutants were eliminated. Strain No. 23, resistant to apramycin and kanamycin B and sensitive to tobramycin, was isolated from soils, identified as Pseudomonas paucimobilis and used as a test organism for overlaying the mutants on agar plate. While inhibition zones were not shown when the parent strains were overlaid with soft agar containing the strain No. 23, clear zones were shown when high tobramycin-producing mutants were overlaid. Using this screening strategy, 58 mutants showing clear zones had been isolated. antibiotic activities of which were incresed to 3~8 fold compared to that of parent strain.

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Mutagenesis of Streptomyces kasugaensis for Kasugamycin Production

  • Cho, Hoon;Choi, Du-Bok;Lim, Chae-Kyu
    • Journal of environmental and Sanitary engineering
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    • v.23 no.4
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    • pp.23-29
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    • 2008
  • This study was performed to develop mutant strain using a combination of UV irradiation procedures with protoplast mutagenesis in order to achieve an effective kasugamycin production from Streptomyceskasugaensis. Whenlessthan 1.0g/lof Linoleic acid was used, the cell growth was not inhibited. On the other hand, the cell growth was greatly inhibited when more than 1.6 g/l of linoleic acid was used. Among the various mutant strains, SK-12 was obtained in medium containing 1.6g/l of linoleic acid, showing the highest rate of both cell growth and kasugamycin production. In order to compare kasugamycin production with the SK-12 and the parent strain using soybean oil, cultures were performed in a flask. The production of kasugamycin was increased with the increase time. The maximum kasugamycin concentration was 1.2g/l after 6 days of culture. The product yield from soybean oil was 0.05g/l/g consumed carbon source, which was roughly 5.0 fold higher than the parent strain. These results show that it was effective method for obtaining a mutant resistant to linoleic acid for the effective production of kasugamycin from soybean oil.