June 2017

New range of Pig P450s Bactosomes

Author:  Isabelle Nobiron, tebu-bio
Posted:  19 June 2017

If you are looking for enzymatic in vitro tools for drug metabolism, reaction phenotyping or metabolite generation using a pig model, you may find these newly released Pig Bactosomes from Cypex of interest.

Bactosomes are recombinant enzymes expressed in E. coli with a patented expression system that enables expression of drug metabolising enzymes without the need for large modifications to be made to the proteins. They demonstrate excellent batch-to-batch consistency and robust activity levels. Each batch is characterized for linearity with time and CYP concentration, Km and Vmax.

The 4 newly released pig Bacterial membranes contain the pig cytochrome P450s CYP1A2 (CYP452), CYP2C49 (CYP451), CYP2E1 (CYP454); CYP3A39 (CYP453), each co-expressed with pig NADPH-cytochrome P450 reductase.

The data shown below are taken from the CYP2E1 characterization as an example:

Typical Batch Characteristics
P450 Concentration: 7 nmol/ml
Protein Concentration: 14 mg/ml
Specific P450 Content: 500 pmol/mg protein
Cytochrome c Reductase Activity: 80 nmol/min/mg protein

View linearity with CYP concentration for pig CYP2E1BR
View linearity with time for pic CYP2E1BR

Typical P450 Activity Data
Activity: 7-ethoxyresorufin O-deethylase
Linear with time up to: 4 min
Linear with P450 up to: 12 pmol/ml
Vmax: 0.6 pmol/min/pmol CYP
Km: 0.03 µM

The CYP2E1 is high reductase supplemented with purified human cytochrome b5 to boost its activity activity. b5 was added to the prep to allow reduction of the Km for substrates (while with some other CYPs, b5 increases the Vmax without affecting the Km). A high reductase CYP3A39 supplemented with cytochrome b5 is to be released soon.

Originally published in tebu-bio's Being Bioreactive
Sekisui XenoTech is the exclusive North American distributor of CYPEX's recombinant enzymes. To purchase in the EU, please visit tebu-bio's website.

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