Functional analysis and its application of multifunctional cytochrome P450 enzymes forming carboxyl group

Research Project

Project/Area Number	26460132
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Natural medicines
Research Institution	Toho University
Principal Investigator	ANZAI Yojiro 東邦大学, 薬学部, 教授 (20318299)
Project Period (FY)	2014-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000) Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywords	抗生物質 / P450酵素 / 酸化反応 / 応用微生物
Outline of Final Research Achievements	The multifunctional cytochrome P450 enzyme RosC catalyzes a two-step, hydroxylation and alcohol oxidation, oxidation reaction to form the C-20 formyl group in the biosynthesis of a 16-membered macrolide antibiotic rosamicin produced by Micromonospora rosaria IFO13697. In this study, we confirmed that RosC has another oxidation activity, which a formyl group at C-20 of rosamicin is converted into a carboxyl group. Moreover, random mutation analysis was also performed for rosC gene using error-prone PCR, and Leucine-176 of RosC is one of amino acid residue controlling of oxidation reaction.