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Review
. 2018 Mar 5;38(2):BSR20171290.
doi: 10.1042/BSR20171290. Print 2018 Apr 27.

Recombinant production of eukaryotic cytochrome P450s in microbial cell factories

Johanna Hausjell  1 , Heidi Halbwirth  1 , Oliver Spadiut  2
Affiliations
Review

Recombinant production of eukaryotic cytochrome P450s in microbial cell factories

Johanna Hausjell et al. Biosci Rep. .

Abstract

Cytochrome P450s (P450s) comprise one of the largest known protein families. They occur in every kingdom of life and catalyze essential reactions, such as carbon source assimilation, synthesis of hormones and secondary metabolites, or degradation of xenobiotics. Due to their outstanding ability of specifically hydroxylating complex hydrocarbons, there is a great demand to use these enzymes for biocatalysis, including applications at an industrial scale. Thus, the recombinant production of these enzymes is intensively investigated. However, especially eukaryotic P450s are difficult to produce. Challenges are faced due to complex cofactor requirements and the availability of a redox-partner (cytochrome P450 reductase, CPR) can be a key element to get active P450s. Additionally, most eukaryotic P450s are membrane bound which complicates the recombinant production. This review describes current strategies for expression of P450s in the microbial cell factories Escherichia coli, Saccharomyces cerevisiae, and Pichia pastoris.

Keywords: Cytochrome P450; Escherichia coli; Pichia pastoris; Saccharomyces cerevisiae; recombinant protein.

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Conflict of interest statement

The authors declare that there are no competing interests associated with the manuscript.

Figures

Figure 1
Figure 1. Exemplary reaction catalyzed by a P450
11-desoxycorticosterone is hydroxylated at position 11 by CYP11B1 leading to corticosterone.
Figure 2
Figure 2. Exemplary crystal structure of a P450
Structure of lanosterol 14α-demethylase (PDB 4LXJ), a P450 crystallized including its membrane anchor (circled in red).
Figure 3
Figure 3. P450 location in a eukaryotic cell
Schematic picture of a eukaryotic cell (left) and the anchoring of an exemplary membrane protein (right) with catalytic domain (red) and its membrane anchor (green).
Figure 4
Figure 4. Strategies for recombinant expression of P450s in E. coli
Overview of strategies applied for the expression of P450s in E. coli.
Figure 5
Figure 5. Interaction of P450 and CPR
Schematic representation of the interaction between P450s (red, PDB 4K9T) and CPRs (green, PDB 3QE2). The CPR reduces NAD(P)H and thereby provides the P450 with electrons for the oxidation of substrates.
Figure 6
Figure 6. Strategies for recombinant expression of P450s in yeasts
Overview of strategies applied for the expression of P450s in yeasts.
See this image and copyright information in PMC

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