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Biotechnology and bioprocess engineering v.15 no.3, 2010년, pp.476 - 480   SCIE 피인용횟수: 1
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Optimization of Lab Scale Methanol Production by Methylosinus trichosporium OB3b

Kim, Hee-Gon    (Department of Environmental Engineering, BK21 Team for Biohydrogen Production, Chosun University   ); Han, Gui-Hwan    (Department of Environmental Engineering, BK21 Team for Biohydrogen Production, Chosun University   ); Kim, Si-Wouk    (Department of Environmental Engineering, BK21 Team for Biohydrogen Production, Chosun University  );
  • 초록

    Methylosinus trichosporium OB3b is a methanotrophic bacterium containing particulate methane monooxygenase (MMO), which catalyzes the hydroxylation of methane to methanol. The methanol is further oxidized to formaldehyde by methanol dehydrogenase (MDH). We developed a novel compulsory circulation diffusion system for cell cultivation. A methane/air mixture (1:1, v/v) was prepared in a tightly sealed gas reservoir and pumped into a nitrate mineral salt culture medium under optimal conditions (5 ${\mu}M$ $CuSO_4$ , pH 7.0, $30^{\circ}C$ ). Cells were harvested, washed, and resuspended (0.6 mg dry cells/mL) in a 500 mL flask in 100 mL of 10 mM phosphate buffer (pH 7.0) containing 100 mM NaCl and 1 mM EDTA as MDH inhibitors, and 20 mM sodium formate. A single 12 h batch reaction at $25^{\circ}C$ yielded a final concentration of 13.2 mM methanol. The use of a repeated batch mode, in which the accumulated methanol was removed after each of three 8 h cycles over a 24 h period, showed a productivity of 2.17 ${\mu}mol$ methanol/h/mg dry cell wt. Finally, a lab-scale reaction performed using a 3 L cylindrical reactor with a working volume of 1 L produced 13.7 mM methanol after 16 h. Our results identify a simple process for improving the productivity of biologically derived methanol and, therefore the utility of methane as an energy source.


  • 주제어

    methane monooxygenase .   methanol dehydrogenase .   Methylosinus trichosporium OB3b .   NaCl .   EDTA.  

  • 참고문헌 (13)

    1. Anthony, C. (1986) Bacterial oxidation of methane and methanol. Adv. Microb. Physiol. 27: 113-210. 
    2. Takeguchi, M., T. Furuto, D. Sugimori, and I. Okura (1997) Optimization of methanol biosynthesis by Methylosinus trichosporium OB3b: An approach to improve methanol accumulation. Appl. Biochem. Biotechnol. 68: 143-152. 
    3. Furuto, T., M. Takeguchi, and I. Okura (1999) Semicontinuous methanol biosynthesis by Methylosinus trichosporium OB3b. J. Mol. Catal. A: Chem. 144: 257-261. 
    4. Lee, S. G., J. H. Goo, H. G. Kim, J. -I. Oh, Y. M. Kim, and S. W. Kim (2004) Optimization of methanol biosynthesis from methane using Methylosinus trichosporium OB3b. Biotechnol. Lett. 26: 947-950. 
    5. Mark, A. C., M. H. Kevin, A. P. Roy, and W. J. Colin (1984) The effect of EDTA and related chelating agents on the oxidation of methanol by the methylotrophic bacterium, Methylophilus methylotrophus. Eur. J. Biochem. 138: 611-615. 
    6. Johannes, F. J., H. K. Simon, P. E. J. Verwiel, A. J. Jacob, C. M. Antonia, and A. D. Johannis (1989) On the mechanism of inhibition of methanol dehydrogenase by cyclopropane-derived inhibitors. Eur. J. Biochem. 184: 187-195. 
    7. Shimoda, M. and I. Okura (1991) Selective inhibition of methanol dehydrogenase from Methylosinus trichosporium (OB3b) by cyclopropanol. J. Mol. Catal. 64: L23-L25. 
    8. Kim, H. G. and S. W. Kim (2006) Purification and characterization of a methanol dehydrogenase derived from Methylomicrobium sp. HG-1 cultivated using a compulsory circulation diffusion system. Biotechnol. Bioproc. Eng. 11: 134-139.     
    9. Burrows, K. J., A. Cornish, D. Scott, and I. J. Higgins (1984) Substrate specificities of the soluble and particulate methane mono-oxygenases of Methylosinus trichosporium OB3b. J. Gen. Microbiol. 130: 3327-3333. 
    10. Cox, J. M., D. J. Day, and C. Anthony (1992) The interaction of methanol dehydrogenase and its electron acceptor, cytochrome cL in methylotrophic bacteria. Biochim. Biophys. Acta 1119: 97-106. 
    11. Kim, H. G., G. H. Han, C. Y. Eom, and S. W. Kim (2008) Isolation and taxonomic characterization of a novel type I methanotrophic bacterium. J. Microbiol. 46: 45-50.     
    12. Dales, S. L. and C. Anthony (1995) The interaction of methanol dehydrogenase and its cytochrome electron acceptor. Biochem. J. 312: 261-265. 
    13. Jensen, S., A. Prieme, and L. Bakken (1998) Methanol improves methane uptake in starved methanotrophic microorganisms. Appl. Environ. Microbiol. 64: 1143-1146. 
  • 이 논문을 인용한 문헌 (1)

    1. 2016. "" Journal of microbiology and biotechnology, 26(7): 1234~1241     

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