title: CYP3A4 activity in the presence of organic cosolvents, ionic liquids, or water-immiscible organic solvents.
abstract: P450 enzymes have attracted the attention of chemists for decades because of their impressive ability to catalyze the hydroxylation of inactivated C--H bonds. However, their use for synthesis in aqueous systems is limited. We report here a survey of the activity of purified human CYP3A4 in the presence of organic solvents or ionic liquids. We show that CYP3A4 tolerates only small amounts (<15 %) of water-miscible organic cosolvents or ionic liquids before its activity toward testosterone drops below detection. [BMIM][PF(6)] in a biphasic system was less detrimental to enzyme activity, with 20 % of the activity remaining in the presence of 15 % of this ionic liquid. CYP3A4 activity in the absence of buffer was only >or=10 % in solvents of the alkane series, with a minimum of 0.85 % water, and with the addition of sucrose and testosterone before enzyme lyophilization. Biphasic solvent systems were more promising, with approximately 85 % of the activity retained.
authors.affiliation: Department of Chemistry, McGill University, 801 Sherbrooke Street West, Montréal, Québec, H3A 2K6, Canada.
authors.forename: Amandine
authors.initials: A
authors.lastname: Chefson
authors.forename: Karine
authors.initials: K
authors.lastname: Auclair
mesh_terms: D002021:Buffers; D002244:Carbon / Q000737:chemistry; D002384:Catalysis; D002625:Chemistry, Organic / Q000379:methods; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000502:physiology*; D006801:Humans; D006859:Hydrogen / Q000737:chemistry; D006860:Hydrogen Bonding; D052578:Ionic Liquids; D007477:Ions; D008956:Models, Chemical; D012997:Solvents / Q000737:chemistry; D013739:Testosterone / Q000737:chemistry; D013997:Time Factors; D014867:Water / Q000737:chemistry*
chemical_list: D002021:Buffers; D052578:Ionic Liquids; D007477:Ions; D012997:Solvents; D014867:Water; D013739:Testosterone; D002244:Carbon; D006859:Hydrogen; D003577:Cytochrome P-450 Enzyme System; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A