Journal of Microbiology and Biotechnology

  • 제22권1호
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  • Pages.141-146
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  • 2012
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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Functional Expression and Characterization of Recombinant NADPH-P450 Reductase from Malassezia globosa

  • Lee, Hwa-Youn (Department of Biological Sciences and Center for Biotechnology Research in UBITA (CBRU), Konkuk University) ;
  • Park, Hyoung-Goo (Department of Biological Sciences and Center for Biotechnology Research in UBITA (CBRU), Konkuk University) ;
  • Lim, Young-Ran (Department of Biological Sciences and Center for Biotechnology Research in UBITA (CBRU), Konkuk University) ;
  • Lee, Im-Soon (Department of Biological Sciences and Center for Biotechnology Research in UBITA (CBRU), Konkuk University) ;
  • Kim, Beom-Joon (Department of Dermatology, College of Medicine, Chung-Ang University) ;
  • Seong, Cheul-Hun (College of Pharmacy, Chung-Ang University) ;
  • Chun, Young-Jin (College of Pharmacy, Chung-Ang University) ;
  • Kim, Dong-Hak (Department of Biological Sciences and Center for Biotechnology Research in UBITA (CBRU), Konkuk University)
  • 투고 : 2011.07.26
  • 심사 : 2011.09.17
  • 발행 : 2012.01.28
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초록

Malassezia globosa is a common pathogenic fungus that causes skin diseases including dandruff and seborrheic dermatitis in humans. Analysis of its genome identified a gene (MGL_1677) coding for a putative NADPH-P450 reductase (NPR) to support the fungal cytochrome P450 enzymes. The heterologously expressed recombinant M. globosa NPR protein was purified, and its functional features were characterized. The purified protein generated a single band on SDS-PAGE at 80.74 kDa and had an absorption maximum at 452 nm, indicating its possible function as an oxidized flavin cofactor. It evidenced NADPH-dependent reducing activity for cytochrome c or nitroblue tetrazolium. Human P450 1A2 and 2A6 were able to successfully catalyze the O-deethylation of 7-ethoxyresorufin and the 7-hydroxylation of coumarin, respectively, with the support of the purified NPR. These results demonstrate that purified NPR is an orthologous reductase protein that supports cytochrome P450 enzymes in M. globosa.

키워드

참고문헌

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피인용 문헌

  1. Evaluation of Luminescent P450 Analysis for Directed Evolution of Human CYP4A11 vol.21, pp.6, 2012, https://doi.org/10.4062/biomolther.2013.086
  2. Enhanced Purification of Recombinant Rat NADPH-P450 Reductase by Using a Hexahistidine-Tag vol.27, pp.5, 2012, https://doi.org/10.4014/jmb.1701.01028