• Title/Summary/Keyword: Random mutation

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Development of a Flavor-Enriched Yeast Extract with a High Glutathione Content (글루타치온 고함유 정미성 효모추출물 개발)

  • Bae, In-Young;Koo, Seung-Hyun;Yoo, Hyun-Jae;Kim, Jong-Min;Bae, Hyun-Ah;Jeon, Eun-Jung;Oh, Eon;Lee, Dae-Hee;Hur, Byung-Serk;Lee, Hyeon-Gyu
    • Korean Journal of Food Science and Technology
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    • v.42 no.5
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    • pp.549-553
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    • 2010
  • Yeast strains with good sensory properties were selected, and those yeasts were subjected to mutation to develop high glutathione producing yeasts. In addition, the antiradical activity and flavoring effect of the yeast extract were evaluated. A total of 68 strains were screened, and three strains of Saccharomyces utilis, four strains of Candida utilis, and one strain of Zygosaccharomyces rouxii were selected based on the flavoring effect. Among them, a random mutation was elicited against SEM-Y8, resulting in a high flavoring effect and growth rate. The glutathione production by SEM-Y8 increased 2.0-fold following the mutation, and the DPPH radical quenching effect of the SY8-M2-1-derived extract increased 3.2-fold compared to that of the wild type. The sensory properties of the SY8-M2-1-derived extract were better than those of garlic or onion extract in umami and mouthfulness. Thus, the SY8-M2-1 extract could be used as a functional flavoring material with improved antiradical activity.

Functional roles of glutamic acid E143 and E705 residues in the N-terminus and transmembrane domain 7 of Anoctamin 1 in calcium and noxious heat sensing

  • Choi, Jonghyun;Jang, Yongwoo;Kim, Haedong;Wee, Jungwon;Cho, Sinyoung;Son, Woo Sung;Kim, Sung Min;Yang, Young Duk
    • BMB Reports
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    • v.51 no.5
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    • pp.236-241
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    • 2018
  • Anoctamin 1 (ANO1) is an anion channel that is activated by changes in cytosolic $Ca^{2+}$ concentration and noxious heat. Although the critical roles of ANO1 have been elucidated in various cell types, the control of its gating mechanisms by $Ca^{2+}$ and heat remain more elusive. To investigate critical amino acid residues for modulation of $Ca^{2+}$ and heat sensing, we constructed a randomized mutant library for ANO1. Among 695 random mutants, reduced $Ca^{2+}$ sensitivity was observed in two mutants (mutant 84 and 87). Consequently, the E143A mutant showed reduced sensitivity to $Ca^{2+}$ but not to high temperatures, whereas the E705V mutant exhibited reduced sensitivity to both $Ca^{2+}$ and noxious heat. These results suggest that the glutamic acids (E) at 143 and 705 residues in ANO1 are critical for modulation of $Ca^{2+}$ and/or heat responses. Furthermore, these findings help to provide a better understanding of the $Ca^{2+}$-mediated activation and heat-sensing mechanism of ANO1.

Site-specific Disruption of Glyoxylate Bypass and Its Effect in Lysine-producing Corynebacterium lactofermentum Strain

  • Kim, Youn-Hee;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.315-320
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    • 1996
  • The role of glyoxylate bypass in a lysine-producing Corynebacterium lactofermentum strain was analyzed. Unlike the wild type, the strain expressed enzymes of glyoxylate bypass during growth in the fermentation broth containing glucose as the carbon source. To evaluate the importance of glyoxylate bypass in the strain, we disrupted chromosomal aceA by using a cloned fragment of the gene. Site-specific disruption of aceA which codes for the isocitrate lyase, the first enzyme of the bypass, was confirmed by Southern blot analysis. The aceA mutant strain completely lost isocitrate lyase activity and ability to grow in a minimal medium containing acetate as the sole carbon source. The mutant strain was similar to its parental strain in growth characteristics and produced comparable amounts of lysine in shake flasks containing glucose as the carbon source. The amount of oxaloacetate accumulated in the fermentation medium was similar for both strains, suggesting that expression of glyoxylate bypass does not necessarily lead to the increase in intracellular oxaloacetate. These data clearly demonstrate that glyoxylate bypass does not function as one of the routes of carbon supply for lysine production in the strain. It appears that the leakiness of the glyoxylate bypass in the strain might be the result of a secondary mutation which arose during previous strain development by random mutagenesis.

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Microbial production of coenzyme Q10

  • Suh, Jung-Woo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2006.11a
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    • pp.127-130
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    • 2006
  • Coenzyme Q10(CoQ10) is a biological quinine compound that is widely found in living organisms including yeast, plants, and animals. CoQ10 has two major physiological activities:(a)mitochondrial electron-transport activity and (b )antioxidant activity. Various clinical applications are also available: Parkinson's disease, Heart disease, diabetes. Because of its various application filed, the market size of CoQ10 is continuously expanding all over the world. A Japanese company, Nisshin Pharma Inc. is the first industrial producer of CoQ10(1974). CoQ10 can be produced by fermentation and chemical synthesis. In several companies, these two methods are used for the production of CoQ10:chemical synthesis - Yungjin, Daewoong, Nishin Parma; fermentation - Kaneka, Kyowa, Yungjin, etc. Researchs in microbial production of CoQ10 have several steps: screening of producing microorganisms, strain development, fermentation process, purification process, scale-up process, plant production. Several strategies are available for the strain development : Random mutation and screening, directed metabolic engineering. For the optimization of fermentation process, various conditions (nutrient, aeration, temperature, culture type, etc.) are considered. Purification is one of the most important step because the quality of final products entirely depends on its purity. The production cost will be reduced and the quality of the CoQ10 will be impoved by continuous researches in strain development, fermentation process, purification process.

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P. aeruginosa EMS1의 mutagen 처리를 통한 고기능 유화재 균주의 개발

  • Lee, Geun-Hui;Lee, O-Mi;Kim, Gi-Han;Cha, Mi-Seon;Son, Hong-Ju;Lee, Sang-Jun
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.556-557
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    • 2001
  • This study was performed to improve the efficency of production of biosudactant which were produced by newly screened MNNGCN-Methyl-N-Nitro- Nitrosoguanidine) mutagenized P. aeruginosa EMS1. A culture grown exponentially for $30^{\circ}C$ in trypic soy brotb is adjusted to pH. MNNG is added and incubated in water bath shaker at about 250 ${\sim}$300rpm. After 20 min, is dilutecl into colded trypic soy broth and centrifugation. The cell pellet is resuspended in 50$m{\ell}$ of trypic soy broth. Cultures are grown at $30^{\circ}C$ overnight. cetyltrimethylammonium bromide-metbylene blue agar plate selected dark blue halo colony. Peanut oil, Castor oil, Olive oil, and so on were compared as carbon source of surface tension and emulsifying activity.

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Applications of Transposon-Based Gene Delivery System in Bacteria

  • Choi, Kyoung-Hee;Kim, Kang-Ju
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.217-228
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    • 2009
  • Mobile genetic segments, or transposons, are also referred to as jumping genes as they can shift from one position in the genome to another, thus inducing a chromosomal mutation. According to the target site-specificity of the transposon during a transposition event, the result is either the insertion of a gene of interest at a specific chromosomal site, or the creation of knockout mutants. The former situation includes the integration of conjugative transposons via site-specific recombination, several transposons preferring a target site of a conserved AT-rich sequence, and Tn7 being site-specifically inserted at attTn7, the downstream of the essential glmS gene. The latter situation is exploited for random mutagenesis in many prokaryotes, including IS (insertion sequence) elements, mariner, Mu, Tn3 derivatives (Tn4430 and Tn917), Tn5, modified Tn7, Tn10, Tn552, and Ty1, enabling a variety of genetic manipulations. Randomly inserted transposons have been previously employed for a variety of applications such as genetic footprinting, gene transcriptional and translational fusion, signature-tagged mutagenesis (STM), DNA or cDNA sequencing, transposon site hybridization (TraSH), and scanning linker mutagenesis (SLM). Therefore, transposon-mediated genetic engineering is a valuable discipline for the study of bacterial physiology and pathogenesis in living hosts.

Effects of flaC Mutation on Stringent Response-Mediated Bacterial Growth, Toxin Production, and Motility in Vibrio cholerae

  • Kim, Hwa Young;Yu, Sang-Mi;Jeong, Sang Chul;Yoon, Sang Sun;Oh, Young Taek
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.816-820
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    • 2018
  • The stringent response (SR), which is activated by accumulation of (p)ppGpp under conditions of growth-inhibiting stresses, plays an important role on growth and virulence in Vibrio cholerae. Herein, we carried out a genome-wide screen using transposon random mutagenesis to identify genes controlled by SR in a (p)ppGpp-overproducing mutant strain. One of the identified SR target genes was flaC encoding flagellin. Genetic studies using flaC and SR mutants demonstrated that FlaC was involved in bacterial growth, toxin production, and normal flagellum function under conditions of high (p)ppGpp levels, suggesting FlaC plays an important role in SR-induced pathogenicity in V. cholerae.

Microbial production of coenzyme Q10

  • Suh, Jung-Woo
    • 한국약용작물학회:학술대회논문집
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    • 2006.11a
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    • pp.127-130
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    • 2006
  • Coenzyme Q10(CoQ10) is a biological quinine compound that is widely found in living organisms including yeast, plants, and animals. CoQ10 has two major physiological activities:(a)mitochondrial electron-transport activity and (b)antioxidant activity. Various clinical applications are also available : Parkinson's disease, Heart disease, diabetes. Because of its various application filed, the market size of CoQ 10 is continuously expanding all over the world. A Japanese company, Nisshin Pharma Inc. is the first industrial producer of CoQ10(1974). CoQ10 can be produced by fermentation and chemical synthesis. In several companies, these two methods are used for the production of CoQ10:chemical synthesis - Yungjin, Daewoong, Nishin Parma; fermentation - Kaneka, Kyowa, Yungjin, etc. Researchs in microbial production of CoQ10 have several steps: screening of producing microorganisms, strain development, fermentation process, purification process, scale-up process, plant production. Several strategies are available for the strain development : Random mutation and screening, directed metabolic engineering. For the optimization of fermentation process, various conditions (nutrient, aeration, temperature, culture type, etc.) are considered. Purification is one of the most important step because the quality of final products entirely depends on its purity. The production cost will be reduced and the quality of the CoQ10 will be impoved by continuous researches in strain development, fermentation process, purification process.

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Selection and Agronomics Characterization of Radiation-Induced Variants in Rice (방사선 처리에 의해 유도된 돌연변이 벼의 주요 특징)

  • Lee, In-Sok;Kim, Dong-Sup;Choi, Su-Ryun;Song, Hi-Sup;Lee, Sang-Jae;Lim, Yong-Pyo;Lee, Young-Il
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.227-232
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    • 2003
  • Radiation technique has been used to develope mutant rice. Suwon 345 rice seeds were irradiated with 250 Gy gamma ray. Morphological characteristics of the variants in M$_{8}$ generation were observed and random amplified polymorphic DNA(RAPD) analysis was carried out. Plant height, panicle length, 1,000 grain weight and lodging were very different in mutants compared with donor cultivar. RAPD analysis showed that polymorphic bands were presented in several primers of the mutants. In comparison with original variety, variants were classified into four group through UPGMA analysis. A group has mutation trait in panicle length, B group in plant height and C group in 1,000 grain weight. Among mutants, no. 46 and 147 was ranked as salt tolerance and the malonaldehyde content of these mutants was more increased than that of original variety. Valuable mutants obtained will be useful for developing new cultivars and for studing gene function in molecular level.l.

Prediction of Atomic Configuration in Binary Nanoparticles by Genetic Algorithm (유전알고리즘을 이용한 이원계 나노입자의 원자배열 예측)

  • Oh, Jung-Soo;Ryou, Won-Ryong;Lee, Seung-Cheol;Choi, Jung-Hae
    • Journal of the Korean Ceramic Society
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    • v.48 no.6
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    • pp.493-498
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    • 2011
  • Optimal atomic configurations in a nanoparticle were predicted by genetic algorithm. A truncated octahedron with a fixed composition of 1 : 1 was investigated as a model system. A Python code for genetic algorithm linked with a molecular dynamics method was developed. Various operators were implemented to accelerate the optimization of atomic configuration for a given composition and a given morphology of a nanoparticle. The combination of random mix as a crossover operator and total_inversion as a mutation operator showed the most stable structure within the shortest calculation time. Pt-Ag core-shell structure was predicted as the most stable structure for a nanoparticle of approximately 4 nm in diameter. The calculation results in this study led to successful prediction of the atomic configuration of nanoparticle, the size of which is comparable to that of practical nanoparticls for the application to the nanocatalyst.