Journal of Microbiology

  • 제43권6호
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  • Pages.499-502
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  • 2005
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  • 1225-8873(pISSN)
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  • 1976-3794(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Characterization of Methylophaga sp. strain SK1 Cytochrome $c_L$ Expressed in Escherichia coli

  • Kim, Hee-Gon (Department of Biomaterials Engineering, Chosun University) ;
  • Phan, Trongnhat (Department of Biomaterials Engineering, Chosun University) ;
  • Jang, Tae-Sa (Department of Chemistry, Chosun University) ;
  • Koh, Moon-Joo (Department of Environmental Engineering, Chosun University) ;
  • Kim, Si-Wouk (Department of Environmental Engineering, Gwangju University)
  • 발행 : 2005.12.01
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초록

Methylophaga sp. strain SK1 is a new restricted facultative methanol-oxidizing bacterium that was isolated from seawater. The aim of this study was to characterize the electron carriers involved in the methanol oxidation process in Methylophaga sp. strain SK1. The gene encoding cytochrome $c_L$ (mxaG) was cloned and the recombinant gene was expressed in Escherichia coli $DH5\alpha$ under strict anaerobic conditions. The recombinant cytochrome $c_L$ had the same molecular weight and absorption spectra as the wild-type cytochrome $c_L$ both in the reduced and oxidized forms. The electron flow rate from methanol dehydrogenase (MDH) to the recombinant cytochrome $c_L$ was similar to that from MDH to the wild-type cytochrome $c_L$. These results suggest that recombinant cytochrome $c_L$ acts as a physiological primary electron acceptor for MDH.

키워드

참고문헌

  1. Anthony, C. 1986. Bacterial oxidation of methane and methanol. Adv. Microbial. Physiol. 27, 113-210 https://doi.org/10.1016/S0065-2911(08)60305-7
  2. Dumont, M.E., J.F. Ernst, D.M. Hampsey, and F. Sherman. 1987. Identification and sequence of the gene encoding cytochrome cheme lyase in yeast Saccharomyces cerevisiae. EMBO J. 6, 235-241
  3. Even, M.T., R.J. Kassner, M. Dolata, T.E. Meyer, and M.A. Cusanovich. 1995. Molecular cloning and sequencing of cytochrome c' from the phototrophic purple sulfur bacterium Chromatium vinosum. Biochim. Biophys. Acta 1231, 220-223 https://doi.org/10.1016/0005-2728(95)00101-N
  4. Evers, T.H. and M. Merkx. 2005. Successful recombinant production of Allochromatium vinosum cytochrome c' requires coexpression of cmm genes in heme-rich Escherichia coli JCB712. Biochem. Biophy. Res. Commun. 327, 668-674 https://doi.org/10.1016/j.bbrc.2004.12.062
  5. Karan, E.F., B.S. Russell, and K.L. Bren. 2002. Characterization of Hydrogenobacter thermophilus cytochrome c (552) expressed in the cytoplasm and periplasm of Escherichia coli. J. Biol. Inorg. Chem. 7, 260-272 https://doi.org/10.1007/s007750100292
  6. Kellogg, J.A. and K.L. Bren. 2002. Characterization of recombinant horse cytochrome c synthesized with the assistance of Escherichia coli cytochrome c maturation factors. Biochim. Biophys. Acta 1601, 215-221 https://doi.org/10.1016/S1570-9639(02)00471-5
  7. Koh, M.J., C.S. Kim, Y.A. Kim, H.S. Choi, E.H. Cho, E. Kim, Y.M. Kim, and S.W. Kim. 2002. Properties of electron carriers in the process of methanol oxidation in a new restricted facultative marine methylotrophic bacterium, Methylophaga sp. MP. J. Microbiol. Biotechnol. 12, 476-482
  8. Londer, Y.Y., P.R. Pokkuluri, D.M. Tiede, and M. Schiffer. 2002. Production and preliminary characterization of a recombinant triheme cytochrome $c_{7}$ from Geobacter sulfurreducens in Escherichia coli. Biochim. Biophys. Acta 1554, 202-211 https://doi.org/10.1016/S0005-2728(02)00244-X
  9. Miller, G.T., B. Zhang, J.K. Hardman, and R. Timkovich. 2000. Converting a c-type to a b-type cytochrome: Met61 to His61 mutant of Pseudomonas cytochrome c-551. Biochemistry 39, 9010-9017 https://doi.org/10.1021/bi000232a
  10. Miller, G.T., D.Q. Mackay, M.S. Standley, S.L. Fields, W.M. Clary, and R. Timkovich. 2003. Expression of Pseudomonas stutzeri Zobell cytochrome c-551 and its H47A variant in Escherichia coli. Protein Expr. Purif. 29, 244-251 https://doi.org/10.1016/S1046-5928(03)00065-2
  11. Moore, G.R. and G.W. Pettigrew. 1990. Cytochrome c: Evolutionary, structural and physicochemical aspects. Springer-Verlag, New York
  12. Rumbley, J.N., L. Hoang, and S.W. Englander. 2002. Recombinant equine cytochrome c in Escherichia coli: High level expression characterization, and folding and assembly mutants. Biochemistry 41, 13894-13901 https://doi.org/10.1021/bi026543y
  13. Sambrook. J. and D.W. Russel. 2001. Molecular Cloning: a Laboratory Manual, 3rd ed. Cold Spring Harbor Laboratory Press, New York
  14. Thomas, P.E., D. Ryan, and W. Levin. 1976. An improved staining procedure for the detection of the peroxidase activity of cytochrome P450 on sodium dodecyl sulphate polyacrylamide gels. Anal. Biochem. 75, 168-176 https://doi.org/10.1016/0003-2697(76)90067-1
  15. Ubbink, M., J.V. Beeumen, and G.W. Canters. 1992. Cytochrome c-550 from Thiobacillus versutus: cloning, expression in Escherichia coli, and purification of the heterologous holoprotein, J. Bacteriol. 174, 3707-3714 https://doi.org/10.1128/jb.174.11.3707-3714.1992