The molecular basis and enzyme engineering strategies for improvement of coupling efficiency in cytochrome P450s

  Yu Ji and Shuaiqi Meng in lack-white Copyright: © biotec

Meng, S., Ji, Y., Zhu, L., Dhoke, G. V., Davari, M. D., Schwaneberg, U., Biotechnology Advances,

Rational design of efficient cytochrome P450s with high catalytic efficiency and coupling efficiency

Cytochrome P450 (P450s) are the most versatile biocatalysts in nature. The catalytic ability of these hemoproteins has extraordinary potential for the synthesis of pharmaceuticals, plastics and hormones. However, the industrial application of P450s is limited, and one of the main bottlenecks is their low coupling efficiency to non-natural substrates. The undesirable uncoupling reactions result in the extra consumption of expensive cofactor NAD(P)H, and lead to the accumulation of reactive oxygen species and the inactivation of enzymes and organisms. Using protein engineering methods, these limitations can be overcome by engineering and fine-tuning P450s. A systemic perspective of the enzyme structure and the catalytic mechanism is essential for P450 engineering campaigns for higher coupling efficiency. This review provide an overview on factors contributing to uncoupling and protein engineering approaches to minimize uncoupling and thereby generating efficient and robust P450s for industrials use. Contributing uncoupling factors are classified into three main groups: i) substrate binding pocket; ii) ligand access tunnel(s); and iii) electron transfer pathway(s). Furthermore, we draw future directions for combinations of effective state-of-the-art technologies and available software/online tools for P450s engineering campaigns.

Shuaiqi Meng was supported by a Ph.D. scholarship from the China Scholarship Council (CSC No. 201906880011).

To learn more, please access the full paper on publications and patents and

Meng, S., Ji, Y., Zhu, L., Dhoke, G. V., Davari, M. D., Schwaneberg, U., Biotechnology Advances,

  P450 engineering campaign Copyright: © Biotechnology Advances

Uncoupling reactions restrict the biocatalysis process of cytochrome P450s in industrial applications. A comprehensive understanding used for rational designing P450 variants with high coupling efficiency is crucial to P450 engineering campaigns.