2019 ; Vol.29-1: 44~54
|Author||Hemraj Rimal, Sang-Cheol Yu, Byeongsan Lee, Young-Soo Hong, Tae-Jin Oh|
|Place of duty||Department of Life Science and Biochemical Engineering, SunMoon University, 70 Sunmoon-ro 221, Tangjeong-myeon, Asan-si, Chungnam 31460, Republic of Korea |
|Title||Characterization of Gel16 as a Cytochrome P450 in Geldanamycin Biosynthesis and in-silico Analysis for an Endogenous Electron Transport System|
J. Microbiol. Biotechnol.2019 ;
|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 Å.
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 → FDR6 → FDX4 → Gel16.|
|Key_word||Biosynthetic pathway, cytochrome P450, electron transport system, geldanamycin, in-silico analysis, redox partner|
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