2017 ; Vol.27-5: 956~964
|Author||Qiuping Yao, Li Ma, Ling Liu, Haruo Ikeda, Shinya Fushinobu, Shengying Li, Lian-Hua Xu|
|Place of duty||Ocean College, Zhejiang University, Dinghai, Zhoushan, Zhejiang 316021, P.R. China|
|Title||Hydroxylation of Compactin (ML-236B) by CYP105D7 (SAV_7469)|
J. Microbiol. Biotechnol.2017 ;
|Abstract||Compactin and pravastatin are competitive cholesterol biosynthesis inhibitors of 3-hydroxy-3-
methylglutaryl-CoA reductase and belong to the statin drugs; however, the latter shows
superior pharmacokinetic characteristics. Previously, we reported that the bacterial P450,
CYP105D7, from Streptomyces avermitilis can catalyze the hydroxylation of 1-deoxypentalenic
acid, diclofenac, and naringenin. Here, we demonstrate that CYP105D7 could also catalyze
compactin hydroxylation in vitro. In the presence of both bacterial and cyanobacterial redox
partner systems with an NADPH regeneration system, the reaction produced two
hydroxylated products, including pravastatin (hydroxylated at the C6 position). The steadystate
kinetic parameters were measured using the redox partners of putidaredoxin and its
reductase. The Km and kcat values for compactin were 39.1 ± 8.8 μM and 1.12 ± 0.09 min-1,
respectively. The kcat/Km value for compactin (0.029 min-1·μM-1) was lower than that for
diclofenac (0.114 min-1·μM-1). Spectroscopic analysis showed that CYP105D7 binds to
compactin with a Kd value of 17.5 ± 3.6 μM. Molecular docking analysis was performed to
build a possible binding model of compactin. Comparisons of different substrates with
CYP105D7 were conclusively illustrated for the first time.|
|Key_word||Compactin, CYP105D7, pravastatin, Streptomyces avermitilis, hydroxylation|
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