Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Surface Display of Heme- and Diflavin-Containing Cytochrome P450 BM3 in Escherichia coli: A Whole-Cell Biocatalyst for Oxidation

  • Yim, Sung-Kun (School of Biological Sciences and Technology, Chonnam National University) ;
  • Kim, Dong-Hyun (School of Biological Sciences and Technology, Chonnam National University) ;
  • Jung, Heung-Chae (Systems Microbiology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Pan, Jae-Gu (Systems Microbiology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kang, Hyung-Sik (School of Biological Sciences and Technology, Chonnam National University) ;
  • Ahn, Tae-Ho (Department of Biochemistry, College of Veterinary Medicine, Chonnam National University) ;
  • Yun, Chul-Ho (School of Biological Sciences and Technology, Chonnam National University)
  • Received : 2009.10.03
  • Accepted : 2009.11.19
  • Published : 2010.04.28
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Abstract

Cytochrome P450 enzymes (P450s) are involved in the synthesis of a wide variety of valuable products and in the degradation of numerous toxic compounds. The P450 BM3 (CYP102A1) from Bacillus megaterium was the first P450 discovered to be fused to its redox partner, a mammalian-like diflavin reductase. Here, we report the development of a whole-cell biocatalyst using ice-nucleation protein (Inp) from Pseudomonas syringae to display a hemeand diflavin-containing oxidoreductase, P450 BM3 (a single, 119-kDa polypeptide with domains of both an oxygenase and a reductase) on the surface of Escherichia coli. The surface localization and functionality of the fusion protein containing P450 BM3 were verified by flow cytometry and measurement of enzymatic activities. The results of this study comprise the first report of microbial cell-surface display of a heme- and diflavin-containing enzyme. This system should allow us to select and develop oxidoreductases containing heme and/or flavins into practically useful whole-cell biocatalysts for extensive biotechnological applications, including selective synthesis of new chemicals and pharmaceuticals, bioconversion, bioremediation, live vaccine development, and biochip development.

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References

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