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Mol. Cells 2010; 30(2): 137-141
Published online July 23, 2010
https://doi.org/10.1007/s10059-010-0098-8
© The Korean Society for Molecular and Cellular Biology
Engineering of Flavonoid O-Methyltransferase for a Novel Regioselectivity
Correspondence to : *Correspondence: jhahn@konkuk.ac.kr
An O-methyltransferase isolated from poplar, POMT7, was identified as a flavone 7-O-methyltransferase. In order to generate a mutant of POMT-7 having a novel regioselectivity, we conducted an error-prone polymerase chain reaction. More than 100 mutants were screened and one of the mutants (POMT-M1) Asp257Gly, methylated the 3-hydroxyl group of flavonols in addition to 7-hydrdoxyl group. The mutation changed asparagine residue at the position of 257 into glycine. The kinetic parameters showed that the wild type POMT7 was better activity toward kaempferol and quercetin than the POMT7-M1. Using E. coli transformant expressing POMT7-M1, 58 μM of 3, 7-O-dimethylquercetin and 70 μM of 3, 7-O-dimethylkaempferol from 100 μM of cor-responding substrate were synthesized successfully.
Keywords flavonoid, O-methyltransferase, regioselectivity
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Research Article
Mol. Cells 2010; 30(2): 137-141
Published online August 31, 2010 https://doi.org/10.1007/s10059-010-0098-8
Copyright © The Korean Society for Molecular and Cellular Biology.
Engineering of Flavonoid O-Methyltransferase for a Novel Regioselectivity
Eun Ji Joe, Bong-Gyu Kim, Byoung-Chan An, Youhoon Chong, and Joong-Hoon Ahn*
Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul 143-701, Korea
Correspondence to:*Correspondence: jhahn@konkuk.ac.kr
Abstract
An O-methyltransferase isolated from poplar, POMT7, was identified as a flavone 7-O-methyltransferase. In order to generate a mutant of POMT-7 having a novel regioselectivity, we conducted an error-prone polymerase chain reaction. More than 100 mutants were screened and one of the mutants (POMT-M1) Asp257Gly, methylated the 3-hydroxyl group of flavonols in addition to 7-hydrdoxyl group. The mutation changed asparagine residue at the position of 257 into glycine. The kinetic parameters showed that the wild type POMT7 was better activity toward kaempferol and quercetin than the POMT7-M1. Using E. coli transformant expressing POMT7-M1, 58 μM of 3, 7-O-dimethylquercetin and 70 μM of 3, 7-O-dimethylkaempferol from 100 μM of cor-responding substrate were synthesized successfully.
Keywords: flavonoid, O-methyltransferase, regioselectivity
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