• Title/Summary/Keyword: UV mutagenesis

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Isolation, Identification and Mutant Development of Butanol Tolerance Bacterium (부탄올 내성 미생물의 분리, 동정 및 변이주의 개발)

  • Jung, Hyesook;Lee, Jinho
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.26-32
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    • 2013
  • Butanol-resistant bacteria were isolated from butanol solvent. The cell growth of isolated strains declined with increasing concentrations of butanol, and isolated strain BRS02 displayed more resistance to 12.5 g/L of butanol than other isolated strains. In addition, strain BRS251, which was resistant to even higher concentrations of butanol, was developed by the mutation of BRS02 using UV. BRS251 could grow in LB medium containing up to 17.5 g/L of butanol, 32.5 g/L of propanol, or 6 g/L of pentanol, whereas the control strain Escherichia coli was found to be tolerant to 7.5 g/L of butanol, 20 g/L of propanol, or 2 g/L of pentanol. The isolated BRS02, a Gram(+) bacterium seen to have a cocci form under the microscope, grew in 6.5% NaCl. According to biochemical tests, BRS02 can metabolize and produce acid with D-galactose, D-maltose, D-mannitol, D-mannose, methyl-${\beta}$-Dglucopyranoside, D-ribose, sucrose, or D-trehalose, as carbon sources. Also, this strain showed resistance to bacitracin, vibriostatic agent O/129, and optochin, alongside positive activities for arginine dihydrolase, ${\alpha}$-glucosidase, and urease. The BRS02 strain was identified as Staphylococcus sp. by analyses of the 16S rRNA gene, phylogenetic tree, and biochemical tests.

The Evaluation of UV-induced Mutation of the Microalgae, Chlorella vulgaris in Mass Production Systems (자외선에 의해 유도된 Chlorella vulgaris 돌연변이 균주의 대량 생산 시스템에서의 평가)

  • Choi, Tae-O;Kim, Kyong-Ho;Kim, Gun-Do;Choi, Tae-Jin;Jeon, Young Jae
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1137-1144
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    • 2017
  • The microalgae Chlorella vulgaris has been considered an important alternative resource for biodiesel production. However, its industrial-scale production has been constrained by the low productivity of the biomass and lipid. To overcome this problem, we isolated and characterized a potentially economical oleaginous strain of C. vulgaris via the random mutagenesis technique using UV irradiation. Two types of mass production systems were compared for their yield of biomass and lipid content. Among the several putatively oleaginous strains that were isolated, the particular mutant strain designated as UBM1-10 in the laboratory showed an approximately 1.5-fold higher cell yield and lipid content than those from the wild type. Based on these results, UBM1-10 was selected and cultivated under outdoor conditions using two different types of reactors, a tubular-type photobioreactor (TBPR) and an open pond-type reactor (OPR). The results indicated that the mutant strain cultivated in the TBPR showed more than 5 times higher cell concentrations ($2.6g\;l^{-1}$) as compared to that from the strain cultured in the OPR ($0.5g\;l^{-1}$). After the mass cultivation, the cells of UBM1-10 and the parental strain were further investigated for crude lipid content and composition. The results indicate a 3-fold higher crude lipid content from UBM1-10 (0.3%, w/w) as compared to that from the parent strain (0.1% w/w). Therefore, this study demonstrated that the economic potential of C. vulgaris as a biodiesel production resource can be increased with the use of a photoreactor type as well as the strategic mutant isolation technique.

Selection of a Mutant Strain with High Yield of Cellulose Production Derived from $Acetobacter$ sp. A9 ($Acetobacter$ sp. A9에서 셀룰로오스 생산량이 높은 변이주 선별)

  • Lee, O-Mi;Son, Hong-Joo;Lee, Sang-Joon
    • Korean Journal of Environmental Biology
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    • v.29 no.4
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    • pp.321-325
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    • 2011
  • The mutant strain M6 derived from Acetobacter sp. A9, which produces high levels of the bacterial cellulose derived by random mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine or UV treatment, was selected by a Hestrin and Schramm medium (HSB) plate assay. The characterization of the cellulose production was studied in flask culture to improve the productivity of bacterial cellulose by $Acetobacter$ sp. A9 and mutant strain M6. The yield of cellulose production was superior to mutant M6 than $Acetobacter$ sp. A9. Cellulose was produced 0.12 g $L^{-1}$ by $Acetobacter$ sp. A9 at HS medium and the mutant M6 produced the cellulose 6.95 g $L^{-1}$at HS medium. Strain M6 produced less amount of gluconic acid than A9, thus showing that cellulose production is negatively relted with the gluconic acid production.

Construction and Characterization of Vitreoscilla Hemoglobin (VHb) with Enhanced Peroxidase Activity for Efficient Degradation of Textile Dye

  • Zhang, Zidong;Li, Wei;Li, Haichao;Zhang, Jing;Zhang, Yuebin;Cao, Yufeng;Ma, Jianzhang;Li, Zhengqiang
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1433-1441
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    • 2015
  • Pollution resulting from the discharge of textile dyes into water systems has become a major global concern. Because peroxidases are known for their ability to decolorize and detoxify textile dyes, the peroxidase activity of Vitreoscilla hemoglobin (VHb) has recently been studied. It is found that VHb and variants of this enzyme show great promise for enzymatic decolorization of dyes and may play a role in achieving their successful removal from industrial wastewater. The level of VHb peroxidase activity correlates with two amino acid residues present within the conserved distal pocket, at positions 53 and 54. In this work, sitedirected mutagenesis of these residues was performed and resulted in improved VHb peroxidase activity. The double mutant, Q53H/P54C, shows the highest dye decolorization and removal efficiency, with 70% removal efficiency within 5 min. UV spectral studies of Q53H/P54C reveals a more compact structure and an altered porphyrin environment (λSoret = 413 nm) relative to that of wild-type VHb (λSoret = 406), and differential scanning calorimetry data indicate that the VHb variant protein structure is more stable. In addition, circular dichroism spectroscopic studies indicate that this variant's increased protein structural stability is due to an increase in helical structure, as deduced from the melting temperature, which is higher than 90℃. Therefore, the VHb variant Q53H/P54C shows promise as an excellent peroxidase, with excellent dye decolorization activity and a more stable structure than wild-type VHb under high-temperature conditions.

Cell Fusion of Cellulolytic Fungi, Aspergillus sp. HB1 (섬유소 분해균 Aspergillus sp. HB 1의 세포융합에 관한 연구)

  • Kim, Joo-Ho;Chang, Sung-Yeoul;Choi, Yong-Keel
    • The Korean Journal of Mycology
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    • v.15 no.2
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    • pp.80-86
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    • 1987
  • The author isolated high cellulolytic fungi from natural sources and determined optimal condition of protoplast formation and fusion as fundamental step for improvement of the isolated it's cellulolytic ability. Three different cellulolytic fungi, such as Aspergillus sp., Penicillium sp. and Trichoderma sp., were isolated from soil. Their cellulolytic activities were compared with that of Aspergillus niger which was useful industrially and had cellulase activity. It was Aspergillus sp. that showed the highest activity of all these four fungi. And then it was followed by Penicillium sp., Trichoderma sp., and Aspergillus niger in order. An auxotrophic mutant of Aspergillus sp. was obtained by UV mutagenesis method. Having try to produce protoplast from mycelia, the author found that ${\beta}-glucuronidase$, at pH 6.0, was effective cell-wall lytic enzyme. And the optimal concentration of this enzyme was 5,000 unit/ml. Regeneration rates of wild type, met. auxotroph and arg. auxotroph, in presence of osmotic stabilizer, were 7. 0%, 7. 5% and 5.2%, respectively. PEG with M.W. 6,000 was effective stimulator for protoplast fusion in the concentration of 30% (W IV). In such a condition, we obtained 1.2% cell fusion rate.

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