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Whole-Cell P450 Biocatalysis Using Engineered Escherichia coli with Fine-Tuned Heme Biosynthesis.

Baodong Hu1,2,3,4, Haibo Yu1,2,3,4, Jingwen Zhou1,2,3,4

  • 1Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China.

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|December 16, 2022
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Summary

This study enhances whole-cell biocatalysis by optimizing intracellular heme biosynthesis to boost P450 enzyme activity. Fine-tuning heme levels improves the production of valuable compounds like drugs and natural products.

Keywords:
Escherichia colichemical intermediatescytochrome P450 enzymesdrugsfine-tuned heme biosynthesis systemnatural productswhole-cell biocatalysts

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Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Enzyme Engineering

Background:

  • Whole-cell biocatalysis using P450 enzymes is valuable for synthesizing compounds.
  • Intracellular heme supply is a critical bottleneck limiting P450 enzyme activity in Escherichia coli.

Purpose of the Study:

  • To develop a strategy for enhancing intracellular heme biosynthesis to improve P450 biocatalytic efficiency.
  • To fine-tune heme levels for increased catalytic activity of P450 enzymes.

Main Methods:

  • Compared strategies for improving heme transport versus biosynthesis.
  • Optimized intracellular heme biosynthesis via integrated gene expression and DNA-guided scaffolds.
  • Employed heme-sensitive biosensors and small regulatory RNA systems for fine-tuning heme levels.

Main Results:

  • Demonstrated enhanced catalytic efficiencies for three P450 enzymes (BM3, sca-2, CYP105D7).
  • Successfully synthesized example products: hydroquinone (chemical intermediate), pravastatin (drug), and 7,3',4'-trihydroxyisoflavone (natural product).

Conclusions:

  • Fine-tuning intracellular heme biosynthesis is a generalizable strategy for improving whole-cell biocatalysts.
  • This approach enhances the production of diverse valuable compounds using hemoproteins.