Enzyme Activity Measurement for Cyclohexanone Monooxygenase

Unsurpassed by any others, Creative Enzymes provides innovative enzyme related services with a professional expert team, and offers leading technical support and personalized customer service found nowhere else in the industry. Creative Enzymes is the most experienced institution specialized in enzyme activity assay, and we are proud to offer the catalytic assay for cyclohexanone monooxygenase.

Cyclohexanone monooxygenase (EC 1.14.13.22; CHMO) is a bacterial flavoenzyme that carries out the stereoselective Baeyer-Villiger type oxidation of a wide range of cyclic ketones into lactones. It was first purified from Nocardia globerula CL1 and Acinetobacter sp. NCIMB 9871. Cyclohexanone monooxygenase is a monomeric FAD and NADPH dependent oxidoreductase and the molecular mass of this enzyme is about 59 kDa. Besides its physiological substrate, cyclohexanone monooxygenase can also oxygenate various cyclic ketones (four- to eight-membered rings), aromatic aldehydes, and heteroatom-containing compounds. Due to its broad substrate tolerance and its high enantioselectivity, this enzyme is a potentially useful industrial catalyst and the possibility of using cyclohexanone monooxygenase in synthetic chemistry to produce chiral synthons has aroused much interest.

For many years, the Baeyer–Villiger oxidation (BVO) reaction has been used for the synthesis of esters and lactones from ketones. There is a wide range of synthetic applications which have been developed, including the production of polymers, perfumes, drugs, and other bioactive molecules. Cyclohexanone monooxygenase is an alternative to conventional chemical catalysts as it avoids hazardous reagents like organic peracids, chlorinated solvents, and metals that are otherwise used in chemical Baeyer-Villiger reactions. Thus, cyclohexanone monooxygenase is attracting an ever-increasing interest for being used in biocatalytic processes. To meet this increasing demands, Creative Enzymes provides activity measurement service for both purified native or recombinant enzymes, and cell extracts containing this enzyme. Cyclohexanone monooxygenase activity is routinely assayed at 25°C by monitoring NADPH oxidation at 340nm. Through working closely with our customers, Creative Enzymes has always been able to deliver high-quality products with the shortest lead time to our customers. In the future, Creative Enzymes will continue to support your aim at notable achievements with our specialized services.

Figure: The closed crystal structure of cyclohexanone monooxygenase from Rhodococcus sp. hi-31.
PDB: 3GWD