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

Autor: Meng S; Institute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074 Aachen, Germany., Ji Y; Institute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074 Aachen, Germany. Electronic address: yu.ji@biotec.rwth-aachen.de., Zhu L; National Center of Technology Innovation for Synthetic Biology, Tianjin 300308, China., Dhoke GV; Institute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074 Aachen, Germany., Davari MD; Department of Bioorganic Chemistry, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120 Halle, Germany. Electronic address: Mehdi.Davari@ipb-halle.de., Schwaneberg U; Institute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074 Aachen, Germany; DWI-Leibniz Institute for Interactive Materials, Forckenbeckstraße 50, 52074 Aachen, Germany. Electronic address: u.schwaneberg@biotec.rwth-aachen.de.
Jazyk: angličtina
Zdroj: Biotechnology advances [Biotechnol Adv] 2022 Dec; Vol. 61, pp. 108051. Date of Electronic Publication: 2022 Oct 18.
DOI: 10.1016/j.biotechadv.2022.108051
Abstrakt: Cytochrome P450s are heme-thiolate enzymes that have been broadly applied in pharmaceutical and biosynthesis because of their efficient oxidation at inert carbons. Extensive engineering campaigns are applied to P450s to explore new non-natural substrates and reactions; however, achieving high coupling efficiency is one of the main challenges. 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). Finally, we draw future directions for combinations of effective state-of-the-art technologies and available software/online tools for P450s engineering campaigns.
Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to declare. All co-authors have seen and agree with the contents of the manuscript and there is no financial interest to report. We certify that the submission is original work and is not under review at any other publication.
(Copyright © 2022 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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