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Overexpression of chaperones GroEL/ES from Escherichia coli enhances indigo biotransformation production of cytochrome P450 BM3 mutant

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Abstract

The self-sufficient cytochrome P450 BM3 mutant (A74G/F87V/D168H/L188Q) can serve as a biocatalyst for whole-cell catalysis process of indigo. Nevertheless, the bioconversion yield of indigo is generally low under normal cultivation conditions (37 °C, 250 rpm). In this study, a recombinant E. coli BL21(DE3) strain was constructed to co-express the P450 BM3 mutant gene and GroEL/ES genes to investigate whether GroEL/ES can promote the indigo bioconversion yield in E. coli. The results revealed that the GroEL/ES system could significantly increase the indigo bioconversion yield, and the indigo bioconversion yield of the strain co-expressing P450 BM3 mutant and GroEL/ES was about 21-fold that of the strain only expressing the P450 BM3 mutant. In addition, the P450 BM3 enzyme content and in vitro indigo bioconversion yield were determined to explore the underlying mechanism for the improvement of indigo bioconversion yield. The results revealed that GroEL/ES did not increase indigo bioconversion yield by increasing the content of P450 BM3 enzyme and its enzymatic transformation efficiency. Moreover, GroEL/ES could improve the intracellular nicotinamide adenine dinucleotide phosphate (NADPH)/NADP+ ratio. Given that NADPH is an important coenzyme in the catalytic process of indigo, the underlying mechanism for the improvement of indigo bioconversion yield is probably related to an increase in the intracellular NADPH/NADP+ ratio.

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Funding

This study was funded by National Basic Research Program of China (973 Program) [Grant number 2012CB721103].

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Authors and Affiliations

  1. Department of Biological Sciences and Biotechnology, Jiangxi Agricultural University, NO.1101 Fangzhimin Avenue, Nanchang, 330045, China

    Shuaiying Peng

  2. Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, NO.500 Caobao Road, Shanghai, 200233, China

    Zhongmei Chu, Jianfeng Lu, Dongxiao Li, Shengli Yang & Yi Zhang

  3. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China

    Yonghong Wang

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  1. Shuaiying Peng
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  2. Zhongmei Chu
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  3. Jianfeng Lu
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  4. Dongxiao Li
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  5. Yonghong Wang
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  6. Shengli Yang
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  7. Yi Zhang
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Correspondence to Shuaiying Peng or Yi Zhang.

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Peng, S., Chu, Z., Lu, J. et al. Overexpression of chaperones GroEL/ES from Escherichia coli enhances indigo biotransformation production of cytochrome P450 BM3 mutant. Biotechnol Lett 45, 993–1000 (2023). https://doi.org/10.1007/s10529-023-03397-5

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  • DOI: https://doi.org/10.1007/s10529-023-03397-5

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