Peroxide-dependent oxidation reactions catalyzed by CYP191A1 from Mycobacterium smegmatis

• Published in: Biotechnology letters Vol. 39; no. 8; pp. 1245 - 1252
• Main Authors: Jo, Hye-Yeong, Park, Sun-Ha, Le, Thien-Kim, Ma, Sang Hoon, Kim, Donghak, Ahn, Taeho, Joung, Young Hee, Yun, Chul-Ho
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.08.2017; Springer Nature B.V
• Subjects: Animals; Applied Microbiology; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biochemistry; Biomedical and Life Sciences; Biotechnology; Butyl hydroperoxide; Candida - enzymology; Candida - genetics; Catalysis; Chemical reactions; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; Dealkylation; E coli; Electron transfer; Escherichia coli - genetics; Fatty acids; Fatty Acids - metabolism; Ferredoxin reductase; Fungal Proteins - genetics; Fungal Proteins - metabolism; Humans; Hydrogen peroxide; Hydroxylation; Life Sciences; Metabolites; Microbiology; Mycobacterium smegmatis; Mycobacterium smegmatis - enzymology; Mycobacterium smegmatis - genetics; NADP; Original Research Paper; Oxidation; Oxidation-Reduction; Peroxides; Peroxides - metabolism; Pharmaceutical Preparations - analysis; Pharmaceutical Preparations - metabolism; Putidaredoxin reductase; Rats; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Substrates; Human P450 substrate; Peroxide; Peroxygenase activity; CYP191A1; Cytochrome P450; Fatty acid
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-017-2358-6

Objectives To find the catalytic activities of CYP191A1 from Mycobacterium smegmatis , in which functions of most P450s are unknown, by using a set of reductase systems, peroxides, and various substrates including fatty acids and human drugs. Results CYP191A1 was functionally expressed in Escherichia coli and purified. Its catalytic activities were examined with fatty acids, chromogenic and fluorogenic substrates, and several human P450 substrates, in the presence of six different types of electron transfer systems, such as rat NADPH-P450 reductase, Candida NADPH-P450 reductase, ferredoxin/ferredoxin reductase, putidaredoxin/putidaredoxin reductase, and peroxides (H 2 O 2 and t -butyl hydroperoxide). The reactions catalyzed by CYP191A1 included the hydroxylation and O -dealkylation of several substrates. Conclusions CYP191A1 preferentially catalyzes the peroxide-dependent oxidation of various substrates over the reductase-dependent reaction. Its peroxygenase activity may be used an effective biocatalytic tool to synthesize the metabolites of drugs.