Tuning a bi-enzymatic cascade reaction in Escherichia coli to facilitate NADPH regeneration for ε-caprolactone production

Bioresources and Bioprocessing

Hefeng Chen¹  Jinghui Xiong¹  Shuang Li¹  Min Zhuo¹  Ting Zhou¹  Ran Liu¹  Hao Yu² 

[1] School of Biology and Biological Engineering, South China University of Technology, Higher Education Mega Center, 510006, Guangzhou, China;School of Chemistry and Chemical Engineering, South China University of Technology, 510006, Guangzhou, China;

关键词: Whole-cell biocatalysis;    RBS design;    NADPH regeneration;    Cyclohexanol;    ε-caprolactone;   

DOI  :  10.1186/s40643-021-00370-w

来源: Springer

【 摘 要 】

ε-Caprolactone is a monomer of poly(ε-caprolactone) which has been widely used in tissue engineering due to its biodegradability and biocompatibility. To meet the massive demand for this monomer, an efficient whole-cell biocatalytic approach was constructed to boost the ε-caprolactone production using cyclohexanol as substrate. Combining an alcohol dehydrogenase (ADH) with a cyclohexanone monooxygenase (CHMO) in Escherichia coli, a self-sufficient NADPH-cofactor regeneration system was obtained. Furthermore, some improved variants with the better substrate tolerance and higher catalytic ability to ε-caprolactone production were designed by regulating the ribosome binding sites. The best mutant strain exhibited an ε-caprolactone yield of 0.80 mol/mol using 60 mM cyclohexanol as substrate, while the starting strain only got a conversion of 0.38 mol/mol when 20 mM cyclohexanol was supplemented. The engineered whole-cell biocatalyst was used in four sequential batches to achieve a production of 126 mM ε-caprolactone with a high molar yield of 0.78 mol/mol.

【 授权许可】

CC BY