• Title/Summary/Keyword: NTG (N-methyl-N'-nitro-N-nitrosoguanidine)

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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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Rhizobium meliloti 102F51 Mutants Defective in Heme Synthesis (Heme 합성특성이 다른 Rhizobium meliloti 102F51 Mutant의 선별)

  • 최영주;정원화;김경수;신평균;조무제
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.98-105
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    • 1986
  • Rhizobium meliloti 102 F 51, the symbiotic partner of alfalfa, was mutagenized with N-methyl-N'-nitro-N-nitrosoguanidine (NTG) and UV-irradiation. Three group of mutants which form white, white-pink and red nodules were selected. The adetylene reduction activity, nodulation activity, amount of heme synthesis during the nodulation, and ${\delta}-aminolevulinic$ acid synthetase (ALAS) and ${\delta}-aminolevulinic$ acid dehydratase (ALAD) activities in free living rhizobia and bacteroid states of the each group of mutants were compared. The mutants forming white nodules showed lower acetylene reduction activity compared to those of red nodule forming mutants. The two key enzymes for the heme synthetic pathway, ALAS and ALAD activities of the mutants forming red nodules was much higher than those of the mutants forming white nodules in bacteroid state, however no significant difference was observed in free living state. In the nodules the ALAS was detected only in bacteroid fraction, while ALAD was detected both in bacteroid and plant fraction. ALAS was dramatically increased with the heme synthesis during the nodulation, while ALAD was decreased in plant fraction but slight increase was observed in bacteroid fraction.

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Development of Phage-resistant Mutants from Lactobacillus casei (Lactobacillus casei의 Bacteriophage내성돌연맥리균분리)

  • 강국희;이경화;박기문;유익제;김영창
    • Microbiology and Biotechnology Letters
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    • v.10 no.3
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    • pp.217-222
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    • 1982
  • A lactic starter organism, Lactobaciilus casei YIT 9018 was treated with N-methyl-N'-nitro-N-nitrosoguanidine (NTG) to obtain phage-resistant mutants. Freshly grown cells suspended in citrate buffer were exposed to NTG of 50 g/$m\ell$ for 40 min. Among 88 colonies isolated eight colonies showed distinct resistance to phages isolated previously from milk plants. The eight new colonies showed character similar to the original L. casei except that they responded differently to phage of different sources and thus were designated as eight different mutants of L casei. From the phage resisting toaether with the fermentative ability equivalent to the mother organism the mutants may be considered to be used as starter cultures for fermented milk.

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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.

Production of L-Ornithine by Citrulline Auxotrophic Mutants of Glutamate-Producing Bacteria

  • CHOI, DAE KEON;WUK SANG RYU;BONG HYUN CHUNG;SOO WAN NAM;YOUNG HOON PARK
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.102-107
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    • 1992
  • For the purpose of producing L-ornithine by microbial fermentation, mutant strains were developed from glutamate-producing bacteria by mutagenesis using N-methyl-N'-nitro-N-nitrosoguanidine (NTG) and UV irradiation. Brevibacterium ketoglutamicum BK1046, a L-citrulline auxotroph which is also resistant to arginine hydroxamate (Arghx), was isolated and selected as the best producer of L-ornithine. This strain was capable of producing L-ornithine at a concentration of 24 g/l after 69 hours of cultivation in the 21 jar fermentor. The optimum supplementary level of L-arginine, a substitute for L-citrulline, was found to be about 0.2 g/l in the shake-flask fermentation.

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Studies on Differentiation of Aspergillus nidulans (I) : Characterization of temperature-sensitive mutants defective in differentiation of aspergillus nidulans (Aspergillus nidulans의 분화에 있어 온도 감수성 돌연변이주의 특성)

  • 조남정;강현삼
    • Korean Journal of Microbiology
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    • v.20 no.4
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    • pp.173-182
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    • 1982
  • From FGSC 159 strain of Aspergillus nidulans, temperature sensitive mutants that are defective in growth and differentiation have been isolated by N-methyl-N'-nitroN-nitrosoguanidine (NTG) treatment. The optimum concentration of NTG and incubation time to get the highest mutation frequency was $100{\mu}g$ per ml and 1 hour, respectively. The survival frequency was 1%. Among the isolated mutants, five strains that were affected in early steps of differentiation were selected for further studies and named smK, smY, smB, smF, and smZ. The execution point of each mutant was determined and the growing pattern of each mutant at the restrictive temperature was observed under the microscope. Growth of mutant was arrested near at the execution point. From genetic analysis, each temperature-sensitive mutants was thought to have a single recessive gene. The genes of smK, smY, smB, smF, and smZ are linked to the chromosome VII, IV, VIII, I, and VI, respectively. It can be concluded that the genes controlling the differentiation are widely dispersed in the genome. From the results of mutant, smK, it is considered that a single gene can affect a function (functions) which act(s) at two different steps during differentiation.

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Studies on the Induction of Available Mutant of Acetic Acid Bacteria by UV light Irradiation and NTG Treatmeat. -On the Organic Acids Composition of Apple Wine Vinegar- (Acetobacter sp.와 그 변이주를 이용한 식초산 발효에 관한 연구 -사과식초의 유기산 조성에 대하여-)

  • 김찬조;박윤중;이석건;오만진
    • Microbiology and Biotechnology Letters
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    • v.9 no.3
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    • pp.139-143
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    • 1981
  • In order to investigate the changes of organic acid contents during the process of apple vinegar, this experiment was conducted by innoculating apple juice with Sarcharomyces cerevisae, and then the apple vinegar were prepared with Acetobacter. aceti and its mutants obtained by the treatment of ultraviolet light and N-methyl-N-nitro-N'-nitrosoguanidine. The organic acids were analyzed by gas chromatography. The contents of malic acid, citric acid and acetic acid in apple juice were 0.73 %, 0.038 % and 0.067%, malic acid, lactic acid and acetic acid in the apple wine 0.114%, 0.10%, and 0.03%, while acetic acid and malic acid in apple vinegar, 4.3 %, and about 0.05 %, respectively.

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Isolation of Mutants Overproducing Amylase from Nuruk Fungi by NTG (NTG에 의한 Amylase활성이 높은 누룩사상균의 변이주의 분리)

  • 정혁준;김영숙;유대식
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.6
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    • pp.987-994
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    • 2000
  • Aspergillus coreanus NR 15-1, Asp. oryzae NR 15-3 and Asp. oryzae NR 2-5 isolated from traditional Korean nuruk were screened as parental strains producing starch hydrolyzing enzymes. They were mutagenized by N-methyl -N'-nitro-N-nitrosoguanidine (NTG) and mutants were isolated for analysis of various amylase activities and the ability of acid production. Among them, the mutants harboring high saccharogenic activity, dextrinogenic activity, and the ability of acid production were selected. Fifteen, six, and five strains of mutants were isolated from Asp. coreanus NR 15-1, Asp. oryzae NR 2-5, and Asp. oryzae NR 15-3, respectively followed by NTG mutagenesis. Among these mutants, thirteen strains were identified as auxotrophic mutants. \ulcorner (Arg. ̄) mutant from Asp. coreanus NR 15-1 showed high glucoamylase activity and total acid productivity. Z6 (Ade. ̄) mutant from Asp. oryzae NR 2-5 showed the highest $\alpha$-amylase activity, therefore \ulcorner and Z6 mutant were selected.

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Characterization of the Mutant of Streptomyces sp. SL-387(KCTC 0102BP) Producing Aminopeptidase M Inhibitors (Aminopeptidase M 저해제를 생산하는 Streptomyces sp. SL-387 (KCTC 0102BP) 변이주의 특성)

  • Chung, Myung-Chul;Chun, Hyo-Kon;Lee, Ho-Jae;Lee, Choong-Hwan;Kho, Yung-Hee
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.47-52
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    • 1995
  • Since the original productivity of new aminopeptidase M inhibitors MR-387A and B by Streptomyces sp. SL-387 (KCTC 0102BP) was not enough for further chemical and biological evaluation, mutation of parent strain by the treatment of N-methyl-N'-nitro-N-nitrosoguanidine was performed in order to obtain a clone with greater inhibitory activity. Mutant N-3 was selected due to a 6-fold greater productivity (40 $\mu$g/ml) than that of the wild type(6.7 $\mu$g/ml). This mutant was resistant to 3,4-dehydro-DL-proline, an antimetabolite of proline, with 25 $\mu$g/ml of minimum inhibitory concentration. Furthermore, the characteristic morphological change from spiral spore chain in wild type to straight in mutant was observed. An aminopeptidase M nhibitor different from MR-387A and B was isolated from the culture broth of the mutant. This inhibitor was composed of 2 proline, 1 valine, and an unknown amino acid which is presumably 3-amino-4-phenylbutanoic acid. IC$_{50}$ value (89.1 $\MU$g/ml) of the purified inhibitor was lower than that of other inhibitors, which may be due to the absence of 2(S)-hydroxyl group within the structure of 3-amino-4-phenyl- butanoic acid.

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Strain Improvement by Interspecific Protoplast Fusion of Streptomyces griseus and Streptomyces hygroscopicus producing Acetaminophen (이종간 원형질체 융합을 이용한 acetaminophen 생산균주 개량)

  • Sohn, Yeo-Won;Jung, Dae-Young;Lee, Sang-Sup;Min, Hong-Ki
    • YAKHAK HOEJI
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    • v.38 no.5
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    • pp.595-601
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    • 1994
  • Acetaminophen, a widely used analgesic, can be formed by N-acetylation and p-hydroxylation of aniline. Interspecific protoplast fusion technique was used to get acetaminophen directly from aniline and to increase the productivity of acetaminophen. Three auxotrophic mutants were obtained from S. griseus(ATCC 13273) and S. hygroscopicus(KCTC 1089) by N-methyl-N'-nitro-N-nitrosoguanidine(NTG) treatment. Regeneration frequencies of S. griseus$(his^-)$, S. griseus$(lys^-)$, S. hygroscopicus$(arg^-)$ were 42%, 45%, and 31%, respectively. Fusion of protoplasts carrying different auxotrophic markers was achieved by treatment with polyethylene glycol. When protoplasts were treated with 50% polyethylene glycol for 3 minutes, the fusion frequency between S. griseus$(his^-)$ and S. hygroscopicus$(arg^-)$ was $3.8{\times}10^{-5}$. The fusion frequency between S. griseus$(lys^-)$ and S. hygroscopicus$(arg^-)$ was $5.6{\times}10^{-4}$. When we checked the production of acetaminophen, thirty-four out of the fifty-six fusants produced larger amounts of acetaminophen than the parent strains did. Nine fusants produced twice more and twenty-five fusants produced one to two times more of acetaminophen than their parents.

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