Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2023 Jan 17;62(2):253-261.
doi: 10.1021/acs.biochem.2c00336. Epub 2022 Aug 31.

Engineering Catalytically Self-Sufficient P450s

Hans Renata  1
Affiliations
Review

Engineering Catalytically Self-Sufficient P450s

Hans Renata. Biochemistry. .

Abstract

The P450 superfamily comprises some of the most powerful and versatile enzymes for the site-selective oxidation of small molecules. One of the main drawbacks for the applications of the P450s in biotechnology is that the majority of these enzymes is multicomponent in nature and requires the presence of suitable redox partners to support their functions. Nevertheless, the discovery of several self-sufficient P450s, namely those from Classes VII and VIII, has served as an inspiration for fusion approaches to generate chimeric P450 systems that are self-sufficient. In this Perspective, we highlight the domain organizations of the Class VII and Class VIII P450 systems, summarize recent case studies in the engineering of catalytically self-sufficient P450s based on these systems, and outline outstanding challenges in the field, along with several emerging technologies as potential solutions.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
P450 catalytic cycle featuring the generation of compound I as the active oxidizing species.
Figure 2.
Figure 2.
Domain organizations of Class VII–X P450s and their comparison to a prototypical multi-component P450 system.
Scheme 1.
Scheme 1.
Oxidative decarboxylation of stearic acid with OleT-BM3R chimera.
Scheme 2.
Scheme 2.
Comparison of several self-sufficient or partially sufficient P450cam fusions for the production of 5-exo hydroxycamphor.
Scheme 3.
Scheme 3.
Comparison of PikC reconstitution with spinach Fdr/Fdx and fusion with RhFRed in the hydroxylation of YC-17 and narbomcyin.
Scheme 4.
Scheme 4.
Screening of several PtmO5 chimeras for the C11 hydroxylation of ent-kaurenoic acid and the synthetic application of PtmO5-RhFRed in the chemoenzymatic synthesis of fischericin B.
Scheme 5.
Scheme 5.
Oxidative biaryl couplings catalyzed by P450 chimeras from RhFRed fusion.
Scheme 6.
Scheme 6.
A. Investigation of CYP725A4-TcCPR fusion for the production of taxadien-5α-ol. B. In vitro conversion of (–)-matairesinol to (–)-pluviatolide with CYP719A23-ATR2 fusion and its comparison with the non-fused counterpart. C. Production of genistein and daidzein with IFS1-CPR chimera construct “E3”, generated by fusion of IFS1 with Catharanthus CPR, followed by N-terminal engineering.
See this image and copyright information in PMC

References

    1. Poulos TL; Johnson EF Structures of Cytochrome P450 Enzymes, in Cytochrome P450: Structure, Mechanism, and Biochemistry, ed. Ortiz de Montellano PR, (Springer, Switzerland, 2015) p. 3–32.
    1. Klingenberg M Pigments of rat liver microsomes. Arch. Biochem. Biophys 1958, 75, 376–386. - PubMed
    1. Guengerich FP Human Cytochrome P450 Enzymes, in Cytochrome P450: Structure, Mechanism, and Biochemistry, ed. Ortiz de Montellano PR, (Springer, Switzerland, 2015) p. 523–786.
    2. Auchus RJ; Miller WL P450 Enzymes in Steroid Processing, in Cytochrome P450: Structure, Mechanism, and Biochemistry, ed. Ortiz de Montellano PR, (Springer, Switzerland, 2015) p. 851–881.
    1. Podust LM; Sherman DH Diversity of P450 enzymes in the biosynthesis of natural products. Nat. Prod. Rep 2012, 29, 1251–1266. - PMC - PubMed
    2. Rudolf JD; Chang C-Y; Ma M; Shen B Cytochromes P450 for natural product biosynthesis in Streptomyces: sequence, structure, and function. Nat. Prod. Rep 2017, 34, 1141–1172. - PMC - PubMed
    3. Zhang X; Guo J; Cheng F; Li S Cytochrome P450 enzymes in fungal natural product biosynthesis. Nat. Prod. Rep 2021, 38, 1072–1099. - PubMed
    1. Fasan R Tuning P450 enzymes as oxidation catalysts. ACS Catal 2012, 2, 647–666.
    2. Urlacher VB; Girhard M Cytochrome P450 monooxygenases in biotechnology and synthetic biology. Trends Biotechnol 2019, 37, 882–897. - PubMed
    3. Girvan HM; Munro AW Applications of microbial cytochrome P450 enzymes in biotechnology and synthetic biology. Curr. Opin. Chem. Biol 2016, 31, 136–145. - PubMed
    4. Jung ST; Lauchli R; Arnold FH Cytochrome P450: taming a wild type enzyme. Curr. Opin. Biotechnol 2011, 22, 809–817. - PMC - PubMed

Publication types

MeSH terms

Substances