CypExpress

CypExpress™ provides for robust metabolite identification and production, while reducing the time and expense involved in this critical process.

Important features:

•    Coexpress a specific human Cyp AND human P450 oxidoreductase

•    Easy to handle dry powder

•    Stable at room temperature

•    CypExpress™ retains catalytic activity for many hours, even overnight

•    Indeed, CypExpress™ can be reused for multiple reaction cycles

•    Addition of Glucose-6-Phosphate Dehydrogenase is not required

•    Reactions can be performed in Tris or Phosphate buffers

•    Added NADP+ & Glucose-6-Phosphate are not essential (but will boost activity)

•    After the reaction, CypExpress™ can be pelleted at relatively low speeds

•    Clean HPLC profiles for easy metabolite ID and purification

•    Very high percent conversions can be obtained from high concentrations of substrate – making purification easier

•    Highly controlled production process for lot-to-lot reproducibility

Using proprietary processes, CypExpress™ are expressed in Pichia yeast. Following optimal expression, the cells are inactivated, permeabalized and dried to a fine powder. CypExpress™ retains the cellular mechanisms to provide the P450 enzymes with the energy and cofactors to continue to function for long experiments, and can generate much larger amounts of metabolite than mammalian microsomes or other genetically engineered expression systems.

To use, just add CypExpress™ to your reaction, mix and agitate (e.g. overnight), and then spin down the CypExpress™ and isolate the product(s). The catalytic activity of traditional systems (microsomal or recombinants) is typically depleted within 20-30 minutes, making it difficult to obtain large quantities of metabolites.

Protocols

Metabolite Production Protocol

Posters

Utilizing a Novel Biocatalytic Human P450 System for the Multi-cycle Production of Metabolites. E. Martinez, et al. Experimental Biology, Boston, MA, USA, 2015.

Multi-cycle Production of Metabolites With Human P450 Systems. E. Martinez, et al., Great Lakes Drug Discovery and Disposition. Ann Arbor, MI, USA, 2015.

A Novel, Robust Recombinant Human P450 Biocatalytic System. E. Martinez, et al., MichBio Drug Discovery and Development. Lansing, MI, USA, 2015

Facile Scale-up Biocatalytic Production of P450 Metabolites Using a Novel Recombinant Human Catalytic System.  E. Martinez, et al., International Society for the Study of Xenobiotics. Orlando, FL, USA, 2015.

Evaluation of Dry Powder Preparations of Permeabilized Pichia pastoris Cells Expressing Human Cytochrome P450 2D6 Enzyme (CYP2D6) for Drug Metabolite Generation. S Li, et al. Gordon Research Conference on Drug Metabolism, Holderness, NH, USA, 2013.

Rapid Identification and Production of Metabolites Using Stabilized Dried Powder (SDP) of Human Cytochrome P450s (hCYPS) 2D6, 3A4, and 2C9 Engineered in Yeast. S Das and M Subramanian. Great Lakes Drug Metabolism and Disposition Group, Indianapolis, IN, USA, 2014.

Rapid Identification and Production of Metabolites Using CypExpressTM 2D6, 3A4 and 2C9. M Subramanian, et al. International Society for the Study of Xenobiotics, San Francisco, CA, USA, 2014.

Cost Comparison