• Title/Summary/Keyword: Mutant strain

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Enhanced Lycopene Production in Recombinant Escherichia coli by Random Transposon and NTG Mutagenesis (Transposon 및 NTG 돌연변이를 이용한 재조합 대장균의 라이코펜 생산성 증진)

  • Yoon, Sang-Hwal;Ko, Min-Su;Park, Kyoung-Ae;Jung, Kyung-Hwa;Shin, Yong-Chul;Lee, Young-Mi;Lee, Sook-Hee;Kim, Seon-Won
    • KSBB Journal
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    • v.21 no.2
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    • pp.90-95
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    • 2006
  • Escherichia coli harboring pAC-LYCO4 and pDdxs was used for lycopene production. Three wild type strains of E. coli OW1, MG1655, and W3110 were compared with DH5${\alpha}$ used before for lycopene production. Lycopene productivity of E. coli MG1655 was similar to DH5${\alpha}$ and the highest among those wild type strain. Therefore, MG1655 strain was used for random transposon and NTG mutagenesis to increase lycopene productivity. Through transposon mutation, five transposon mutants with increased lycopene productivity were obtained. It was found that genes knocked out by transposon insertion were treB in Tn1 mutant, B2436 in Tn2 mutant, and rfaH in Tn3, 4, and 5 mutants. Lycopene productivity was the highest in Tn4 mutant among the Tn mutants, which was 6-fold and 8-fold higher in lycopene concentration and content, respectively, in comparison with those obtained with wild type strain. NTG4 mutant was acquired with NTG mutation. The highest lycopene productivity of 6 mg/L and 4 mg/g DCW was obtained from the NTG4 mutant when arabinose of 0.013 mM was added for induction of dxs, rate-limiting gene of MEP pathway. The lycopene productivity of NTG4 mutant was increased 18-fold and 12-fold in lycopene concentration and content, respectively when comparing with the wild type strain.

Production of Bluish Purple Pigment from Streptomyces californicus KS-89 (Streptomyces californicus KS-89 변이주에 의한 청자색소의 생산)

  • 이병호;이상훈
    • KSBB Journal
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    • v.9 no.2
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    • pp.147-156
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    • 1994
  • A study was carried out for production of a pigment : bluish purple, using a mutant Streptomyces californicus KS-89-7. The mutant was induced from Streptomyces californicus KS-89 with N-methyl-N-nitro-N-nitrosoquanidin(MNNG). It was immobilized on an inert substance made of colloidal sillica and 3.5% sodium alginate with 1 to 10 ratio. The diameter of inert bead was 2mm, and number of immobilized mutant spore was approximately $1.0{\times}10^7$/ml. It was packed in a column reactor and fermentation was conducted with a substrate made of soluble starch 1%, glycerol 1.0%, sodium glutamate 0.1%, sodium nitrate 0.05%, L-prolin 0.025% and with some trace elements. The aeration for production of the pigment was 2.5m1/min with semi-continuous fermentation. The pigment production reached at peak on 8 days of fermentation, and the mutant produced the pigment 1.8 times more than its parent strain with the maximum pigment production of $1.72g/\ell$. The pigment production continued for 24 hours of fermentation, and at the end of the fermentation the mutant produced the pigment $1.52g/\ell$.

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Role of HtrA in growth of Streptococcus mutans under acidic environment (산성환경에서 S. mutans의 생육에 미치는 HtrA gene의 영향)

  • Kang, Kyung-Hee
    • Proceedings of the KAIS Fall Conference
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    • 2009.12a
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    • pp.498-500
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    • 2009
  • 본 연구에서는 한국인 아동의 우식치아로부터 분리한 S. mutans K7으로부터 HtrA gene을 동정하고 HtrA expression이 산성환경하에서 S. mutans의 생육에 미치는 영향을 알아보았다. S. mutans K7의 HtrA mutant strain은 산성환경에서 parental strain과 비교하였을 때, 생육에 있어서 상당한 차이를 나타내었다. 또한 Biofilm formation에 관여하는 GtfB, 와 GtfC의 발현량도 현저히 줄어들었다. 그리고 HtrA mutant strain에 HtrA gene을 삽입하여 HtrA의 발현량을 회복하였을 경우에는 acid stress하에서 control과 같은 nomal growth phenotype을 회복하였다. 이러한 결과들은 S. mutans K7에서 HtrA가 acid stress동안에 중요한 역할을 담당함을 제시하고 있다.

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Enhanced Alcohol Production from Synthesis Gas Using a CO-resistant Mutant of Clostridium sp. AWRP (CO 내성을 갖는 Clostridium sp. AWRP 돌연변이주의 분리 및 이를 이용한 합성가스로부터의 에탄올 생산 연구)

  • Kwon, Soo Jae;Lee, Joungmin;Lee, Hyun Sook
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.581-584
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    • 2019
  • In this study, the carbon monoxide (CO)-fermenting acetogen, Clostridium sp. AWRP was subjected to chemical mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine (NTG) to generate a CO-resistant mutant. Among the 26 colonies obtained, the highest alcohol production was observed in one isolate, named C1. Compared to the wild-type strain, the C1 strain exhibited 1.5- and 3.4-fold higher CO consumption rate and alcohol selectivity, respectively. The total CO consumption of strain C1 could be further enhanced by increasing the content of metal ions, such as nickel and iron. The highest ethanol titer (5.7 g/l) was achieved by 5-fold increase in the iron concentration.

High Production of L-Crnithine by L-Citrulline Auxotroph of Brevibacterium ketoglutamicum : PART I : Selection of Stable Mutant Strains with Less Revertant Formation (Brevibacterium ketoglutamicum을 이용한 L-Ornithine 생산 연구 PART I : L-Ornithine 생산 고역가 안정화 균주 선발)

  • 류욱상;장형욱;이홍원;정준기;장순재;유연우;박영훈
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.292-297
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    • 1999
  • Overproduction of L-ornithine by mutant strains isolated from Brevibacterium detoglutamicum BK1046 was investigated. The strain was a L-ornithine auxotroph and exhibited culture instability during fermentation. Through a sequential screening effort, a highly stable strain with lless revertant formation was finally selected and designated B. ketoglutamicum BK52 (KCTC0141BP). It prouduced L-ornithine at a high concentration (above 9 g/L) independent of subculture or cultivation time, and also had a very low tendency of revertant formation. In a long-term storage, this strain maintained its cell stability and productivity of L-ornithine to a reasonable range.

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Isolation and Its Optimal Culture Condition for High Agarase-Producing Mutant (한천분해효소의 고생산성 변이주의 분리 및 최적배양조건)

  • 황선희;하순득;김봉조;김학주;공재열
    • KSBB Journal
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    • v.14 no.3
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    • pp.351-357
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    • 1999
  • A marine bacterium Bacillus cereus ASK202, agarase producing strain, was treated with some mutagenic agents, ultraviolte(UV), 1-methyl-3-nitro-1-nitrosoguanidine(NTG), and ethyl methane sulfonate(EMS), several times for the increasing of the agarase production After mutagen treatment, we isolated one mutant strain treated with NTG showed the highest stability and agarase productivity and named as Bacillus cereus ASK202-N3. This Bacillus cereus ASK202-N3 strain was well grown in the modified marine medium containing 0.5%(w/v) agar, 0.3%(w/v) yeast extract, and 5.0%(w/v) NaCl, and the optimal initial pH, temperature and culture time were 7.8, $25^{\circ}C$ and 32h, respectively. In the optimal culture conditions, the agarase production was increased to 5.3 fold(850units/L) compared to that of the wild type.

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Isolation of a Tryptophan-Overproducing Strain Generated by EMS Mutagenesis of Candida rugosa

  • Kim, Sun-Hye;Ha, Yu-Mi;Youm, Hyoung-Joon;An, Gil-Hwan;Lee, Bong-Duk;Won, Mi-Sun;Song, Kyung-Bin
    • Preventive Nutrition and Food Science
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    • v.9 no.2
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    • pp.187-189
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    • 2004
  • To isolate a mutant strain which overproduces tryptophan, mutants of Candida rugosa were screened after EMS (ethyl methane sulfonate) mutagenesis. Fluorotryptophan, a tryptophan analogue, was used for selection of a tryptophan-overproducing mutant after mutagenesis. Among 50 mutants, several candidates were selected based on intracellular tryptophan content. Amino acid analysis results showed that C3 was the best strain because it had the highest amount of tryptophan among the mutants.

Isolation and characteristics of yellow-pigment producing mutants of Monascus anka. (Monascus anka로부터 황색소 생성 변이주의 분리 및 특성)

  • 이호재;이형주
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.111-115
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    • 2002
  • To produce yellow pigment selectively, mutants were induced from Monascus anka Nakazawa et Sato IFO 4478 (KCCM 11832 strain), and their characteristics were evaluated. Five kinds of auxotrophic mutants which required amino acids for growth and pigmentation, were isolated through a series of mutagenic treatments. Especially, asparagine auxotroph Y7 produced high ratio of yellow pigment. This mutant showed all the morphological characteristics of Monascuceae but the shape of colony and the diameter of conidia. Mutant Y7 was propagated by sexual reproduction more often than asexual reproduction, which could be effective in production of pigments. Yellow pigment produced extracellularly by the mutant Y7 was more soluble in polar solvents such as ethanol and water than in nonpolar solvents. Its productivity of yellow pigment was 2.2 times higher in the mutant Y7 than in parents. In addition, its yellow pigment showed characteristics of maximum absorption at 373 nm. Moreover, the hue of pigment produced by the mutant Y7 was bright yellow, and it was stable through the subculture over 10 generations.

Relationship of the Redox State of Pyridine Nucleotides and Quinone Pool with Spectral Complex Formation in Rhodobacter sphaeroides 2.4.1 (Rhodobacter sphaeroides 2.4.1 내의 pyridine nucleotide와 quinone pool의 redox 상태와 광합성기구의 합성과의 상관관계)

  • Ko, In-Jeong;Oh, Jeong-Il
    • Journal of Life Science
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    • v.19 no.7
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    • pp.852-858
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    • 2009
  • The homeostasis of the pyridine nucleotide pool [NAD(P)H and NAD(P)$^+$] is maintained in Rhodobacter sphaeroides mutant strains defective in the cytochrome bci complex or the cytochrome c oxidases in terms of its concentration and redox state. Aerobic derepression of the puf operon, which is under the control of the PrrBA two-component system, in the CBB3 mutant strain of R. sphaeroides was shown to be not the result of changes in the redox state of the pyridine nucleotides and the ubiquinone/ubiquinol pool. Using the bc$_1$ complex knock-out mutant strain of R. sphaeroides, we clearly demonstrated that the inhibitory effect of cbb$_3$, oxidase on spectral complex formation is not caused indirectly by the redox change of the ubiquinone/ubiquinol pool.

Characterization of Mutant Strain, Lactobacillus paracasei ML-7 Isolated from Kimchi, and Its Effect on the Growth of Broiler (김치 유래의 유산균 변이주 Lactobacillus paracasei ML-7의 특성 및 육계 성장에 대한 영향)

  • Park, Keun-Tae;Oh, Mihyang;Nam, Jungok;Ji, Kibbeum;Han, Jong-Kwon
    • Korean Journal of Food Science and Technology
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    • v.46 no.2
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    • pp.148-152
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    • 2014
  • A mutant strain of Lactobacillus paracasei (ML-7) was derived by ultraviolet irradiation of the parent L. paracasei LS-2 strain, followed by colony selection on pH 3.5 MRS agar plates, on which L. paracasei LS-2 cannot grow. The L. paracasei ML-7 mutant strain produced an increased amount of organic acids compared to the L. paracasei LS-2 parent strain. Broiler chickens were randomly fed with one of three experimental diets: a basal diet as a control, and basal diets supplemented with 0.3% of the L. paracasei LS-2 and L. paracasei ML-7 strains. When concluding the study, the final body weights of the broilers fed with the supplemented basal diets were found to be significantly higher (1.9% for LS-2 and 6.2% for ML-7) than the control broiler group. Broilers fed with basal diet supplemented with the mutant L. paracasei ML-7 strain showed increased body weight and food intake, compared to broilers fed with the basal diet with or without the parent L. paracasei LS-2 strain.