Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characterization of Gel16 as a Cytochrome P450 in Geldanamycin Biosynthesis and in-silico Analysis for an Endogenous Electron Transport System

  • Rimal, Hemraj (Department of Life Science and Biochemical Engineering, Sun Moon University) ;
  • Yu, Sang-Cheol (Department of Life Science and Biochemical Engineering, Sun Moon University) ;
  • Lee, Byeongsan (Chemical Biology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Hong, Young-Soo (Chemical Biology Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Oh, Tae-Jin (Department of Life Science and Biochemical Engineering, Sun Moon University)
  • Received : 2018.09.07
  • Accepted : 2018.11.01
  • Published : 2019.01.28
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Abstract

Geldanamycin and its derivatives, inhibitors of heat shock protein 90, are considered potent anticancer drugs, although their biosynthetic pathways have not yet been fully elucidated. The key step of conversion of 4,5-dihydrogeldanamycin to geldanamycin was expected to catalyze by a P450 monooxygenase, Gel16. The adequate bioconversions by cytochrome P450 mostly rely upon its interaction with redox partners. Several ferredoxin and ferredoxin reductases are available in the genome of certain organisms, but only a few suitable partners can operate in full efficiency. In this study, we have expressed cytochrome P450 gel16 in Escherichia coli and performed an in vitro assay using 4,5-dihydrogeldanamycin as a substrate. We demonstrated that the in silico method can be applicable for the efficient mining of convenient endogenous redox partners (9 ferredoxins and 6 ferredoxin reductases) against CYP Gel16 from Streptomyces hygroscopicus. The distances for ligand FDX4-FDR6 were found to be $9.384{\AA}$. Similarly, the binding energy between Gel16-FDX4 and FDX4-FDR6 were -611.88 kcal/mol and -834.48 kcal/mol, respectively, suggesting the lowest distance and binding energy rather than other redox partners. These findings suggest that the best redox partners of Gel16 could be NADPH ${\rightarrow}$ FDR6 ${\rightarrow}$ FDX4 ${\rightarrow}$ Gel16.

Keywords

Acknowledgement

Supported by : National Research Foundation of Korea (NRF), Korea Polar Research Institute

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