Progress towards the easier use of P450 enzymes

Abstract

The cytochrome P450 enzymes (P450s or CYPs) form a large family of heme proteins involved in drug metabolism and in the biosynthesis of steroids, lipids, vitamins and natural products. Their remarkable ability to catalyze the insertion of oxygen into non-activated C-H bonds has attracted the interest of chemists for several decades. Very few chemical methods exist that directly hydroxylate aliphatic or aromatic C-H bonds, and most of them are not selective or of limited scope. Biocatalysts such as P450s represent a promising alternative: however, their applications have been limited by substrate specificity, low activity, poor stability and the need for cofactors. This review covers the attempts to overcome these limitations using approaches such as mutagenesis, chemical modifications, conditions engineering and immobilization.

Efficient Screening of Cytochrome P450 BM3 Mutants for Their Metabolic Activity and Diversity toward a Wide Set of Drug-Like Molecules in Chemical Space

“…Because it has been shown that drug metabolites produced by P450s may have improved pharmacological activities and are often responsible for the toxicity or unwanted pharmacological side effects, there is increasing interest in systems enabling the facile (bio)synthesis of sufficient quantities of metabolites for structural elucidation and pharmacological and toxicological evaluation (Chefson and Auclair, 2006). Biocatalytic metabolite production can be performed by large-scale incubations with recombinant human P450s.…”

Towards Preparative Scale Steroid Hydroxylation with Cytochrome P450 Monooxygenase CYP106A2

“…They are involved in the synthesis of secondary metabolites like vitamins and steroids as well as in detoxification of xenobiotics. [1] Because they enable the insertion of oxygen (hydroxylation, epoxidation) into aromatic and aliphatic, activated and nonactivated CÀH bonds, typically in a regio-and stereoselective manner, P450s are of tremendous interest for the synthesis of fine chemicals and pharmaceuticals. [2] Due to their oxygen and cofactor dependency, limited stability and activity and the low water solubility of typical substrates these enzymes are mainly restricted to lab-scale applications.…”

Detection of 25-Hydroxyvitamin D3 with an Enzyme modified Electrode

“…The human CYP27B1 enzyme has been expressed in E. coli, partially purified and its activity has been demonstrated using LC-MS/ MS. A glassy carbon electrode modified with this enzyme, a non-native redox mediator Co(sep) 3+ , and pH-adjusted Nafion ® has been shown to respond to the circulating form of vitamin D, 25(OH)D 3 . Using cyclic and square wave voltammetry, the GCE/Nafion ® /Co(sep) 3+ /CYP27B1 electrode has been shown to be successful in detecting 25(OH)D 3 in buffer within the physiological range (5-200 ng/ml).…”

“…The proof-of-concept developed in this study demonstrates that the detection of 25(OH)D 3 in buffer is possible, although behavior of the modified electrode across various 25(OH)D 3 concentrations has yet to be determined. The sensor is sensitive to 25(OH)D 3 in the physiological range (5-200 ng/ml) although optimization of the electrode assembly and mode of detection will be required in order to operate the sensor in the entire range.…”

“…These enzymes are capable of metabolizing a diverse range of chemicals using different biotransformation reactions and are associated with biosynthesis of steroids, vitamins and lipids, xenobiotic and drug metabolism [1][2][3][4][5][6][7][8]. Thus, there has been immense biotechnological interest in these enzymes, and they have been utilized for applications in bioelectronic devices, biochips, bioreactors, and biosensor technologies [9][10][11][12].…”