• Title/Summary/Keyword: Auxotroph

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Genotoxicity Test of Bojungikkeehapdaechilki-tang water extract (보중익기합대칠기탕(補中益氣合大七氣湯) 추출물의 유전독성 평가)

  • Hwang, Hui-Jeung;Byun, Joon-Seok;Heo, Jin-Il
    • Herbal Formula Science
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    • v.14 no.1
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    • pp.141-167
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    • 2006
  • The genotoxicity of water extract of Bojungikkeehapdaechilki-tang was tested by In Vitro Chromosome Aberration Test. Bacterial Reverse Mutation Assay and Micronucleus test according to OECD Guidelines and KFDA Guidelines. The obtained results were as follows : 1. Chromosome Aberration Test: In Vitro Chromosome Aberration Test of Bojungikkeehapdaechilki-tang extracts was carried out using cultured Chinese hamster lung cells in the presence and absence of metabolic activation system(S-9 mix). No significant changes in the number of aberrant metaphases having structural and number of aberrations were detected in Bojungikkeehapdaechilki-tang extracts treated groups. 2. Bacterial Reveres Mutation Assay: Bojungikkeehapdaechilki-tang extracts was evaluated for its potential to induce reverse mutation in the histidine auxotroph strains of Salmonella typhimurium such as TA100, TA1535, TA98 and TAl537 and the tryptophan auxotroph strain of Escherichia coli WP2 uvrA. No significant changes in the number of revertant colonies compared to its negative control were detected in Bojungikkeehapdaechilki-tang extracts treated groups against all 5 strains. 3. Micronucleus test: Micronucleus test of Bojungikkeehapdaechilki-tang extracts were performed using specific pathogen free 7-week old male ICR mouse. No significant changes in the number of micronucleated polychromatic erythrocytes among 2000 polychromatic erythrocytes compared to negative control were detected in all Bojungikkeehapdaechilki-tang extracts treated groups. In summarized above-mentioned results, it is concluded that Bojungikkeehapdaechilki-tang extracts have not genotoxicity against In Vitro Chromosome Aberration Test, Bacterial Reverse Mutation Assay and Micronucleus test.

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Production of L-Tyrosine by PFP Resistant Mutant Induced from Brevibnrcterium sp. (Brevibacterium sp. 로부터 유도된 PFP 내성 변이주에 의한 L-Tyrosine 생성)

  • Bae, Jun-Tae;Park, Gyeong-Suk;Lee, Byeol-Na
    • The Korean Journal of Food And Nutrition
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    • v.9 no.1
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    • pp.21-28
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    • 1996
  • This study was attempted to investigate the production of L-tyrosine by Brevibacterium flavum ATCC 14067. To select the strain which produce more L-tyrosine, mutants were induced by N-methyl-N'-nitro-nitrosoguanidine (NTG) treatment and phenylalanine auxotrophic mutants were induced by NTG and penicillin treatments. PFP resistant mutant was isolated from a phenylalanine auxotroph by retreatment with NTG and screened for increase of L-tyrosine production. PFP-326 mutant resistant to PPP (100ug/ml) was derived from phenylalanine auxotroph by mutagenesis with NTG and PFP-106 mutant resistant to PFP (1201g/ml) was derived from PFP-326 by mutagenesis with NTG. The composition of media for L-tyrosine production in strain PFP-106 was studied. PFP-106 mutant strain produced 50mg 11 of L-tyrosine while the parent strain produced 0.56mg 11 of L-tyrosine. The optimum composition of medium for L-tyrosine by strain PFP-106 was 10cA sucrose as carbon source, 3% ammonium sulfate as nitrogen source. The optimum cultural condition for producing L-tyrosine by strain PFP-106 was L-phenylalanine at a concentration of 1000g/mg.

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국내기탁기관의 현황 2

  • 오두환
    • The Microorganisms and Industry
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    • v.15 no.1
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    • pp.38-42
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    • 1989
  • Industrial strain Improvement is concerned with developing or modifying microorga-nisms used In production of commercially important fermentation products. The aim is to reduce the production cost by improving productivity of a strain and manipulating specific cilarafteristic such as the ability to utilize cheaper raw materials or resist bacteriophages. The traditional empiri-cal approach to strain improvement is mutation combined with selection and breeding techniques. It is still used by us to improve the productivity of organisms in amino acids. organic acids andenzymes production. The breeding of high L-lysine-producing strain Au112 is one of the outstanding examples of this approach. It is it homoserine auxotroph with AEC, TA double metabolicanalogue resistant markers. The yield reaches 100g/1. Resides, the citric acid-producing organism Aspergillus nuger, Co827, its productivity reches the advanced level in the world, is also the result of a series mutations expecially with Co Y-radiation. The thermostable a-amylaseroducing strain A 4041 is the third example. By combining physical and chemical multations. the strain ,A 4041becomes an asporogenous, catabolite derepressed mutant with rifamycin resistant and methionine, arginine auxotroph markers. The a-amylase activity reaches 200 units/ml. The fourth successful example of mutation in strain improvement is the glucoamylase-producing strain Aspergillus nigerSP56 its enzyme activity is 20,000 units/ml, 4 times of that of the parental strain UV_11. Recently recombinant DNA approach Provides a worth while alternative strategy to Industrial strain improve-ment. This technique had been used by us to increase the thermostable a-amylase production and on some genetic researches.

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Modigication of host cells and Expression of Recombinant E. coli trp plasmids for the increased Production of Tryptophan in Klebsiella pneumoniae (Klebsiella pneumoniae에서 트립토판 생산증대를 위한 숙주개발 및 재조합 trp plasmid의 발현)

  • 지연태;홍광원;박장현;이세영
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.46-51
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    • 1987
  • In order to increase the production of tryptophan by maximizing expression of recombinant trp plasmid, Klebsiella pneumoniae KC 105(pheA tyrA trpE trpR tyrR) was genetically modified. KC 107, inosine monophospate(IMP) auxotroph from KC 105 and KC 108, histidine(His) auxotroph from KC 107 were also derived respectively to increase phosphoribosylpyrophosphate(PRPP) production which is required for tryptophan biosynthesis. From KC 107 phosphoribosylpyrophosphate consumption which is required for tryptophan biosynthesis. From KC 107 and KC 108, KC 109 and KC 110, both arginine auxotrophs were derived respectively. To investigate the expression of recombinant trp plasmid in the selected K. pneumoniae mutants, the auxotrophic mutants were transformed with recombinant trp plasmids pSC 101-$trpE^{FBR}$, pSC 101-trpL(.DELTA.att) $trpE^{FBR}$ (pSC 101-trp-AF). Amount of tryptophan produced and activities of tryptophan synthase of $trpR^{-}$ mutant (KC 100) and $tyrR^{-}$ mutnat(KC 105) containing recombinant plasmid pSC 101-trp operon were increased by 30-40% as compared with KC 99(pheA tyrA trpE) containing recombinant plasmid pSC 101-trp operon. Activities of tryptophan synthase and production of tryptophan of KC 108 ($His^{-}$) and KC 109($Arg^{-}$) containing recombinant plasmid pSC 101-trp operon were increase by two-fold as compared with KC 107 containing pSC 101-trp operon.

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Isolation of a High-Yield Mutant Strain for L-Proline Production and Its Fermentation Conditions

  • Ryu, Wuk-Sang;Jang, Hyung-Wook;Cho, Kyoung-Hee;Chang, Soon-Jae;Ryu, Yeon-Woo;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.613-618
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    • 1999
  • L-Proline-producing mutant strains were developed by exposing L-glutamic acid-producing bacteria to N-metyl-N-nitro-nitrosoguanidine and UV irradiation. A L-histidine auxotroph of Corynebacterium acetoacidophilum RYU3161(KCTC 0616BP), which was resistant to sulfaguanidine and proline analogs (DHP, AZC, TAC), was isolated. The activity of the mutant strain's $\gamma$-glutamyl kinase was 45% higher than that of the parent strain. The optimum level of L-histidine for production of L-proline was 0.16 g/l. In a 5-1 jar fermenter, the mutant strain produced L-proline at a high concentration (35 g/l) level within 48 h of cultivation.

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Disruption of the metC Gene Affects Methionine Biosynthesis in Pectobacterium carotovorum subsp. carotovorum Pcc21 and Reduces Soft-Rot Disease

  • Seonmi, Yu;Jihee, Kang;Eui-Hwan, Chung;Yunho, Lee
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.62-74
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    • 2023
  • Plant pathogenic Pectobacterium species cause severe soft rot/blackleg diseases in many economically important crops worldwide. Pectobacterium utilizes plant cell wall degrading enzymes (PCWDEs) as the main virulence determinants for its pathogenicity. In this study, we screened a random mutant, M29 is a transposon insertion mutation in the metC gene encoding cystathionine β-lyase that catalyzes cystathionine to homocysteine at the penultimate step in methionine biosynthesis. M29 became a methionine auxotroph and resulted in growth defects in methionine-limited conditions. Impaired growth was restored with exogenous methionine or homocysteine rather than cystathionine. The mutant exhibited reduced soft rot symptoms in Chinese cabbages and potato tubers, maintaining activities of PCWDEs and swimming motility. The mutant was unable to proliferate in both Chinese cabbages and potato tubers. The reduced virulence was partially restored by a complemented strain or 100 µM of methionine, whereas it was fully restored by the extremely high concentration (1 mM). Our transcriptomic analysis showed that genes involved in methionine biosynthesis or transporter were downregulated in the mutant. Our results demonstrate that MetC is important for methionine biosynthesis and transporter and influences its virulence through Pcc21 multiplication in plant hosts.

Transposon Tn5 Mutagenesis of Bradyrhizobium japonicum: A Histidine Auxotrophic Mutant of B. japonicum Shows Defective Nodulation Phenotype on Soybean

  • So, Jae-Seong
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.110-113
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    • 1995
  • Transposon Tn5 was used to induce random insertional mutations in Bradyrhizobium japonicum, a soybean endosymbiont. By genomic Southern blot analysis, transposition events were found to have occurred randomly throughout the B. japonicum genome. After screening 3, 626 mutants by auxotrophy test, a histidine auxotroph was isolated. Upon plant infection test, the His mutant showed a 3~4 day delay in nodule formation.

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Production of 5-IMP by Auxotroph of Brevibacterium ammoniagenes (Brevibacterium ammoniagenes의 영양구성 변리주에 의한 5 -IMP 생성)

  • 이별나
    • The Korean Journal of Food And Nutrition
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    • v.1 no.2
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    • pp.37-42
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    • 1988
  • In attempts to obtain IMP Producting strains, Brevibacterium ammoniagenes ATCC 6872 was treated wilts N.1.6. Adenine-guanine requiring mutants were obtained from Brevibacterium ammoniagenes ATCC 6872, and then a strain of them was selected for production of IMP and named Brevibacterium ammoniagenes No.9(ade', gu'). The production of IMP by Brevibacterium ammoniagenes nuts No.9 was about 3mg/ml for 4 day of culture. The optimal concentration of adenine and guanine was 150mg/mg.

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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.

Genotoxicity Study of HM10411, Recombinant Human Granulocyte Colony Stimulating Factor (재조합 인과립구 콜로니 자극인자 HM10411의 유전독성 연구)

  • 권정;이미가엘;홍미영;조지희;정문구;권세창;이관순
    • Biomolecules & Therapeutics
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    • v.10 no.4
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    • pp.268-273
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    • 2002
  • Mutagenic potential of HM10411 (recombinant human granulocyte colony stimulating factor) was evaluated by bacterial reverse mutation test, in vitro chromosome aberration test and in vivo micronucleus test. The bacterial reverse mutation test was performed using the histidine auxotroph strains of Salmonella typhimurium TA100, TA1535, TA98, TA1537 and tryptophan auxotroph strain of Escherichia coli WP2 uvrA. The negative results of the bacterial reverse mutation test suggest that HM10411 does not induce mutation, in the genome of Salmonella typhimurium and E. coli under the conditions used. In addition, it has little clastogenicity either in vitro chromosome aberration test or in vivo micronucleus test. For in vitro chromosomal aberration test, Chinese hamster lung(CHL) cells were exposed to HM10411 of 23, 46 or 92 $\mu\textrm{g}$/ml for 6 or 24 hours in the absence and for 6 hours in the presence of metabolic activation system. There was no significant increase in the number of aberrant metaphase in HM 10411-treated groups at any dose levels both in the presence and absence of metabolic activation system. The micronucleus test was carried out using specific pathogen free(SPF) 7-week old male ICR mice, The test item, HM10411 was intraperitoneally administered at 1150, 2300 or 4600 $\mu\textrm{g}$/kg once a day for 2 consecutive days. There was no significant increase in the frequencies of micronucleated polychromatic erythrocytes(PCEs) at any treated groups compared with negative control group. Therefore, these results demonstrate that the test item, HM10411, was not mutagenic under the condition of these studies.