Co-expression of Prefoldin From Hyperthermophilic Archaea Pyrococcus furiosus in Escherichia coli Enhances The Catalytic Efficiency of Modified Cytochrome P450 BM3

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Co-expression of Prefoldin From Hyperthermophilic Archaea Pyrococcus furiosus in Escherichia coli Enhances The Catalytic Efficiency of Modified Cytochrome P450 BM3
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Affiliation:State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; Key Laboratory of Synthetic Biology, Chinese Academy of Sciences,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; Key Laboratory of Synthetic Biology, Chinese Academy of Sciences,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; Key Laboratory of Synthetic Biology, Chinese Academy of Sciences,State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; Key Laboratory of Synthetic Biology, Chinese Academy of Sciences,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; Key Laboratory of Synthetic Biology, Chinese Academy of Sciences
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Fund Project:This work was supported by a grant from National Basic Research Program of China (2012CB721103)

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Abstract:

P450 BM3 is derived from Bacillus megaterium, and its mutant (A74G, F87V, L188Q, D168H) could hydroxylate indole to indigo in Escherichia coli whereas the bioconversion yield is low under normal culture conditions (37℃, 250 r/min). Chaperones from hyperthermophilic archaea Pyrococcus furiosus were co-expressed with P450 BM3 mutant in E. coli to investigate whether they could improve the production of indigo, and results indicated that prefoldin could significantly increase the indigo production. Our research also revealed that enzyme activity was not the restrictive factor in the indigo bioconversion process. We also demonstrated that prefoldin could increase the intracellular nicotinamide adenine dinucleotide phosphate (NADPH)/NADP⁺ ratio, and this ratio was probably related to the increase of the indigo bioconversion yield.

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PENG Shuai-Ying, CHU Zhong-Mei, LU Jian-Feng, LI Dong-Xiao, WANG Yong-Hong, YANG Sheng-Li, ZHANG Yi. Co-expression of Prefoldin From Hyperthermophilic Archaea Pyrococcus furiosus in Escherichia coli Enhances The Catalytic Efficiency of Modified Cytochrome P450 BM3[J]. Progress in Biochemistry and Biophysics,2017,44(12):1125-1131

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Received:April 10,2017
Revised:August 23,2017
Accepted:September 04,2017
Online: December 19,2017
Published: December 20,2017

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