Mempro™ Cell-Based Cytochrome P450 Production

Creative Biostructure can offer high-quality custom Mempro™ cytochrome P450 production services based on cell-based expression systems. Our scientists can provide you multiple overexpression and purification strategies for high-yield production of stable and functional membrane proteins.

Mempro™ cell-based protein production systems are popularly used for the production of numerous integral membrane proteins, such as G-protein coupled proteins (GPCRs) and cytochromes P450 (CYPs, hemoproteins) superfamily proteins. CYPs are composed of a superfamily of those proteins containing a hemo cofactor. CYPs use numerous small and large molecules as substrates in enzymatic reactions. The most common reaction catalyzed by CYPs is a monooxygenase reaction. CYPs can be a major source of variability in drug pharmacokinetics and response. Most CYPs locate in the liver and require a protein partner to deliver one or more electrons to reduce the iron. Owing to the the nature of the electron transfer proteins, CYPs can be classified into several groups.

Figure 1. Illustration of the biocatalysis by CYP1A2 and CPR on the E. coli cell surface. (Microb Cell Fact., 2016)

Creative Biostructure can provide various strategies for cytochrome P450 production using Mempro™ cell-based proteins expression systems, including:

• Mempro™ Protein Production in Bacterial Cells System ;
• Mempro™ Protein Production in Yeast Cells System;
• Mempro™ Protein Production in Incest Cells System;
• Mempro™ Protein Production in Mammalian Cells System.

Among Mempro™ cell-based protein production platforms, Escherichia coli (E. coli) is used as bacterial host for the production of cytochromes P450. We can perform two principal strategies in E. coli cells, the expression as soluble membrane proteins inserted into the membrane or as inclusion bodies representing cytoplasmic aggregates. Yeast cells (such as saccharomyces cerevisiae and pichia pastoris) system, combines prokaryotic as well as eukaryotic characteristics. Yeast is an attractive eukaryotic host due to single cells, fast growth rates, inexpensive media, high cell densities. Insect cells (such as Sf9 and Sf21) are also a major system for cytochrome P450 expression, which is developed in the early stage that transfected insect cells with the baculovirus species AcMNPV. Mammalian cells present another eukaryotic expression system, which provide the most authentic cell environment. Mammalian cell lines derived from COS, CHO, BHK-21, HEK293, HeLa and GH3 are commonly used for cytochrome P450 production.

Creative Biostructure is capable of expressing, isolating, purifying and crystallizing cytochromes P450 to facilitate the study of their biological functions. Misfolding, aggregation, inactivity, poor stability and solubility are the common issues encountered in integral membrane protein production. However, these issues can be overcome by optimizing factors like expression vectors, host strains, growth conditions, co-expression of helper proteins and fusion tags, etc.

We provide other various Mempro^TM membrane protein production services. Please feel free to contact us for a detailed quote.

References:
Cytochrome P450. (https://en.wikipedia.org/wiki/Cytochrome_P450).
P. Quehl, et al. (2016). Co-expression of active human cytochrome P450 1A2 and cytochrome P450 reductase on the cell surface of Escherichia coli. Microb Cell Fact., 15(1): 26.
U. M. Zanger, and M. Schwab (2012). Cytochrome P450 enzymes in drug metabolism: Regulation of gene expression, enzyme activities, and impact of genetic variation. Pharmacol. Ther., 138(1): 103-141.