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KMID : 0578320100300020137
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Molecules and Cells
2010 Volume.30 No. 2 p.137 ~ p.141
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Engineering of Flavonoid O-Methyltransferase for a Novel Regioselectivity
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Joe Eun-Ji
Kim Bong-Gyu
An Byoung-Chan
Chong You-Hoon
Ahn Joong-Hoon
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Abstract
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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.
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KEYWORD
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flavonoid, O-methyltransferase, regioselectivity
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