Biochem/physiol Actions

Cytochrome P450 enzymes are heme containing monooxygenases which are found primarily in the mammalian liver. They catalyze the oxidative metabolism of xenobiotics. This metabolism is the initial step in the biotransformation and elimination of a wide variety of drugs and environmental pollutants from the body. Sulfaphenazole, fluconazole, fluvoxamine and fluoxetine are inhibitors, whereas rifampin is an inducer of the enzyme. The isozyme CYP2C9 is a major pathway for the invivo conversion of Diclofenac to 4-Hydroxydiclofenac via hydroxylation.

Cytochrome P450 is a heterogeneous family of isozymes whose primary function is to oxidize small molecules, both as a function of intermediary metabolism (e.g., fatty acids) and to detoxify exogenous compounds (drugs or toxins). Some isoforms have narrow substrate specificity, while others are promiscuous.

Features and Benefits

CypExpress^™ 2C9 is a permeabilized and stabilized dried yeast powder preparation containing full length, unmodified, human CYP 2C9 and recombinant human NADPH oxidoreductase. Using proprietary processes, CypExpress^™ are expressed in Pichia yeast. Following optimal expression, the cells are inactivated, permeabilized 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 larger amounts of metabolite than mammalian microsomes or other genetically engineered expression systems. It is designed for rapid screening of drug metabolites and for scaling up metabolite generation.

Reconstitution

Per Protocol/Variable

Unit Definition

A reaction mixture containing 500 μM diclofenac and 100 mg/mL of CypExpress^™ 2C9 will produce 428 μM 4-hydroxydiclofenac in 2 hours at 30°C under the conditions described.

Physical form

Light yellow, very fine powder

Application

Poster: A Novel, Robust Recombinant Human P450 Biocatalytic System

Poster: Evaluation of Dry Powder Preparations of Permeabilized
Pichia pastoris Cells Expressing Human Cytochrome P450 2D6 Enzyme (CYP2D6) for Drug Metabolite Generation

Poster: Facile Scale-up Biocatalytic Production of P-450 Metabolites Using a Novel Recombinant Human Catalytic System

Poster: Multi-Cycle Production of Metabolites with human P450 systems

Poster: Rapid identifcation and production of metabolites using CypExpress^™ 2D6m 3A4 and 2C9

Poster: Rapid identifcation and production of metabolites using stabilized dried powder (SDP) of human cytochrome P450S (hCYPS) 2D6, 3A4, and 2C9 engineered in yeast

Poster: Utilizing a Novel Biocatalytic Human P450 System for the Multi - Cycle Production of Metabolites

Legal Information

CypExpress is a trademark of Oxford Biomedical Research