• Title/Summary/Keyword: UV mutagenesis

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Isolation of Trp, Thr Overproducing Strain of Saccharomyces cerevisiae (Trp, Thr Analogue 복합 저항성 Saccharomyces cerevisiae 균주 개발)

  • 염형준;이승현;김선혜;선남규;안길환;이봉덕;원미선;송경빈
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.6
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    • pp.1017-1021
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    • 2004
  • To isolate a mutant which overproduces threonine and tryptophan, mutants of Saccharomyces cerevisiae were screened after UV and EMS mutagenesis. Hydroxynorvaline, a Thr analogue was used for selection of a Thr-overproducing mutant after UV mutagenesis. Among 31 mutants, TC 5-1 was selected as the strain candidate, based on amino acid analysis. TC 5-1 was then treated by EMS mutagenesis for Trp overproduction. Eight mutants were selected using fluorotryptophan for Thr and Trp overproducing strains. Amino acid analysis results showed that TC 6-1 was the best strain since it had the highest amount of Thr and Trp among mutants.

Adaptive Responses on Survival and Mutagenesis during MNNG Pretreatmeat and Lethality to UV MNNG at Different Cell Stages in Aspergillus nidulans (Aspergillus nidulans에서 MNNG 선 처리시의 생존도와 돌연변이 유발에 대한 Adaptive response 및 Cell stage 따른 UV와 MNNG에 대한 치사율 조사)

  • Chae, Suhn-Kee
    • The Journal of Natural Sciences
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    • v.9 no.1
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    • pp.45-52
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    • 1997
  • We have examined the effects of low concentrations of MNNG, alkylating agent, in survival and mutagenesis in Aspergillus nidulans. Pretreatments of cells with nontoxic and submutagenic doses of MNNG did not reduce the cytotoxic and mutagenic effects of exposure to a high concentration of drug. The results imply that adaptive responses on survival and mutagenesis for MNNG treatments do not occur in Aspergillus nidulans. In the first mitotic cell cycle during germination, the sensitivity to MNNG has been investigated at hourly time interval, and compared with that for UV irradiation. In both UV and MNNG treatments, the sensitivity increased till S cell stage, and decreased while DNA replication continued. Different from that show for UV irradiation, lethality to MNNG reached to the maximum at G2 cell stage.

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Gene-Specific Repair of 6-4 Photoproducts in Trichothiodystrophy Cells

  • Nathan, Sheila;Van Hoffen, Anneke;Mullenders, Leon H.F.;Mayne, Lynne V.
    • BMB Reports
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    • v.32 no.6
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    • pp.554-560
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    • 1999
  • TTD1BI cells are non-hypersensitive to UV irradiation and perform normal genome repair of pyrimidine dimers but fail to excise 6-4 photoproducts and, concomitantly, are unable to restore RNA synthesis levels following UV irradiation. This pointed to a detect in gene-specific repair and this study was undertaken to examine repair of 6-4 photoproducts at the gene-level. The results indicated a defect in gene-specific repair of 6-4 photoproducts in active genes, although strand-specificity of 6-4 photoproduct removal was essentially similar to that of normal cells. These findings indicate that the near normal UV resistance of TTD1BI cells may be due to the inability of these cells to remove DNA lesions preferentially, as well as to the cells opting out of the cell cycle to repair damage before resuming replication.

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Biosorption of uranium by Bacillus sp.FB12 isolated from the vicinity of a power plant

  • Xu, Xiaoping;He, Shengbin;Wang, Zhenshou;Zhou, Yang;Lan, Jing
    • Advances in environmental research
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    • v.2 no.3
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    • pp.245-260
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    • 2013
  • Biosorption represents a technological innovation as well as a cost effective excellent remediation technology for cleaning up radionuclides from aqueous environment. In the present study, a bacteria strain FB12 with high adsorption rate of uranium ion was isolated from the vicinity of the nuclear power plant. It was tentatively identified as Bacillus sp.FB12 according to the 16S rDNA sequencing. Efforts were made to further improve the adsorption rate and genetic stability by UV irradiation and UV-LiCl cooperative mutagenesis. The improved strain named Bacillus sp.UV32 obtains excellent genetic stability and a high adsorption rate of 95.9%. The adsorption of uranium U (VI) by Bacillus sp.UV32 from aqueous solution was examined as a function of metal ion concentration, cell concentration, adsorption time, pH, temperature, and the presence of some foreign ions. The adsorption process of U (VI) was found to follow the pseudo-second-order kinetic equation. The adsorption isotherm study indicated that it preferably followed the Langmuir adsorption isotherm. The thermodynamic parameters values calculated clearly indicated that the adsorption process was feasible, spontaneous and endothermic in nature. These properties show that Bacillus sp.UV32 has potential application in the removal of uranium (VI) from the radioactive wastewater.

Strain Improvement of Streptomyces minoenisis DMCJ-144, An ${\alpha}$-Amylase Inhibitor Producing Actinomycetes (알파-아밀라제 저해제 생산 방선균 Streptomyces minoensis DMCJ-144의 균주개량)

  • 최응칠;김숙경;강동희;이재우;김병각
    • Biomolecules & Therapeutics
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    • v.1 no.1
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    • pp.26-30
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    • 1993
  • Strain of treptomyces minoensis DMCJ-144 was tried to be improved so that it produces much more the $\alpha$-amylase inhibitor. Streptomyces minoensis DMCJ-144 was treated with 1 mg/mι (pH 9.0) of N-methyl-N'-nitro-N-nitrosoguanidine at $30^{\circ}C$ for 60 min and irradiated with UV light distanced 30 cm for 20 min. After mutagenesis, surviving colonies were cultured on the CM contaning acriflavine ($10{\mu}g/ml$) three times in order to enhance the mutability. And then through multi-level screening, colonies that ${\alpha}$-amylase inhibitor productibility. was Improved were selected by modified-blue value method. After third acriflavine treatment, $\alpha$-amylase inhibitory activities of selected colonies were found to be much better as compared with that of parent strain. One mutant strain showed 5.4 time inhibitory activity than the parent strain.

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Strain Improvement of Candida tropicalis for the Production of Xylitol: Biochemical and Physiological Characterization of Wild-type and Mutant Strain CT-OMV5

  • Rao Ravella Sreenivas;Jyothi Cherukuri Pavanna;Prakasham Reddy Shetty;Rao Chaganti Subba;Sarma Ponnupalli Nageshwara;Rao Linga Venkateswar
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.113-120
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    • 2006
  • Candida tropicalis was treated with ultraviolet (UV) rays, and the mutants obtained were screened for xylitol production. One of the mutants, the UV1 produced 0.81g of xylitol per gram of xylose. This was further mutated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and the mutants obtained were screened for xylitol production. One of the mutants (CT-OMV5) produced 0.85g/g of xylitol from xylose. Xylitol production improved to 0.87 g/g of xylose with this strain when the production medium was supplemented with urea. The CT-OMV5 mutant strain differs by 12 tests when compared to the wild-type Candida tropicalis strain. The XR activity was higher in mutant CT-OMV5. The distinct difference between the mutant and wild-type strain is the presence of numerous chlamvdospores in the mutant. In this investigation, we have demonstrated that mutagenesis was successful in generating a superior xylitol-producing strain, CT-OMV5, and uncovered distinctive biochemical and physiological characteristics of the wild-type and mutant strain, CT-OMV5.

Combination Strategy to Increase Cyclosporin A Productivity by Tolypocladium niveum Using Random Mutagenesis and Protoplast Transformation

  • Lee, Mi-Jin;Duong, Cae Thi Phung;Han, Kyu-Boem;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.869-872
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    • 2009
  • The cyclic undecapeptide cyclosporin A (CyA), one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. To increase CyA productivity by wild-type T. niveum (ATCC 34921), random mutagenesis was first performed using an antifungal agar-plug colony assay (APCA) selection approach. This generated a mutant strain producing more than 9-fold greater CyA than the wild-type strain. Additionally, a foreign bacterial gene, Vitreoscilla hemoglobin gene (VHb), was transformed via protoplast regeneration and its transcription was confirmed by RT-PCR in the UV-irradiated mutant cell. This led to an additional 33.5% increase of CyA production. Although most protoplast-regenerated T. niveum transformants tend to lose CyA productivity, the optimized combination of random mutagenesis and protoplast transformation described here should be an efficient strategy to generate a commercially valuable, yet metabolite low-producing, fungal species, such as CyA-producing T. niveum.

Screening of Phytase Overproducing Strains in Aspergillus spp. by UV Mutagenesis

  • Lee, Eung-Suek;Paik, In-Kee;Hahm, Young-Tae
    • Mycobiology
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    • v.28 no.3
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    • pp.119-122
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    • 2000
  • Phytases (myo-inositol hexakisphosphate phosphohydrolase; EC 3.1.3.8) are enzymes which catalyze the hydrolisys of phytate into myo-inositol and inorganic phosphates. Phytases are found in plants and a variety of microorganisms. Aspergillus species were treated with 254 nm of UV irradiation for the screening of phytase overproducing mutant strains. At 15 minute irradiation, the survivals of population were less than 5%, and UV irradiation time was decided at 20 minute for the isolation of mutant strains. Four UV mutant strains in A. oryzae (YUV-47, -169, -341, -511) and six in A. ficuum (FUV-17, -36, -69, -193, -317, -419) were isolated on PSM media containing ammonium phosphate. The specific enzyme activities of A. ficuum mutants are 110 to 140% higher than that of wild type.

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Function of muc Gene on Mutagenesis and DNA Repair (돌연변이와 DNA 손상회복에 미치는 muc 유전자의 기능)

  • 전홍기;이상률;백형석
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.192-198
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    • 1990
  • To determine whether the muc region of pKM101 and its mutant pSL4 is sufficient for the expression of these phenotypes, muc regions of pKM101 and pSL4 were subcloned onto the highcopy number vector pKB354 and were selected the cloned pJB200 and pJB210. The recombinant plasmids pJB200 and pJB210 were introduced into umu $C36^{-}$ uvr $A6^{-}$ (TK610) strain and determined the protection effect and mutagenecity for UV and MMS. The protection effect and mutagenecity of umu $C36^{-}$ uvr $A6^{-}$ (TK610) were supressed by muc gene of the recombinant plasmids. The muc gene of pSL4 has higher effect than that of pKM101. The recombiant plasmid pJB210(inclued muc gene of pKM101) did not affect uv-mutagenesis in the $recA^{-}$ (JC2926) mutant.

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Strain Improvement of Penicillium verruculosum for High Cellulase Production by Induced Mutation (섬유소분해효소 생산증진을 위한 Penicillium verruculosum의 균주개량)

  • 정기철
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.388-395
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    • 1987
  • In order to obtain a regulatory mutant strain with high cellulase activity, a newly isolated Penicillium verrculosum, strain F-3 was used as parental strain since it was proved to be an efficient cellulase producer. A number of experiments were conducted to determine the optimum conditions to in-duce mutagenesis and isolate the desirable mutant strains. Out of several restriction compounds tested, 1.5% oxgall was found to be most effective to restrict the colony size by suppressing overgrowth. Derepression of catabolites was employed as a criterion in selecting mutant strains with high cellulase productivity. Production of cellulase by Penicillium venculosum F-3 was suppressed when cultured on the media with more than 1% of glucose or glycerol. It was found that either irradiation with UV light for 19 mins or treatment with nitrosoguanidine at 200$\mu\textrm{g}$/m1 for 60 mins, induced mutagenesis at desired level, when the survival rate of the spore was 0.2% and 48%, respectively. Three mutant strains of F-3, UV-9, UV-10, and NTG-3 that had the highest cellulase productivity were finally selected, based on filter paper degradation rate, size of clearing zone on the screening plate and cellulase activity in the medium containing cellulose powder. When the mutant strains were compared with parental strain F-3, on the KC-M-W medium containing cellulose powder, the filter paper activities of UV-9, UV-10, and NTG-3 were increased by 34%, 55%, and 41%, respectively. However, the assimilation of cellobiose octaacetate by UV-9 or NTG-3 was markedly reduced. When the mutant UV-10 was grown on cellobiose octaacetate medium (CCA-4) in shaking flasks, the cellulase activities of the mutant increased by 20 to 50% compared to the parental strain. Excreation of soluble protein from the mutant also elevated up to 30%. The mutant also constitutively produced both CMCase and $\beta$-glucosidase, though at relatively low level, in the presence of glucose or cellobiose as carbon sources.

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