Purification of 2-hydroxyisoflavanone dehydratase from the cell cultures of Pueraria lobata

被引:37
作者
Hakamatsuka, T
Mori, K
Ishida, S
Ebizuka, Y
Sankawa, U
机构
[1] Univ Tokyo, Grad Sch Pharmaceut Sci, Bunkyo Ku, Tokyo 1130033, Japan
[2] Toyama Med & Pharmaceut Univ, Fac Pharmaceut Sci, Toyama 9300194, Japan
关键词
Pueraria lobata; Leguminosae; isoflavone biosynthesis; 2-hydroxy-isoflavanone; daidzein; dehydratase; cell cultures;
D O I
10.1016/S0031-9422(98)00266-0
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
2-Hydroxyisoflavanone dehydratase, which catalyzes the final step of the formation of the isoflavonoid skeleton, was purified and characterized from yeast extract-elicited cell suspension cultures of Pueraria lobata. 2-Hydroxyisoflavanone, the substrate of the dehydratase, is the product of 2-hydroxyisoflavanone synthase, as cytochrome P-450 which catalyzes the hydroxylation step associated with aryl migration of flavanone. The dehydratase was purified to apparent homogeneity for the first time by a seven-step purification procedure. It is a single polypeptide with a molecular weight of 38 kDa, and has an isoelectric point at pH 5.1 and a pH optimum at 6.8. It required no co-factor, and the apparent Michaelis constant for 2,7,4'-trihydroxyisoflavanone was 7.0 mM. (C) 1998 Published by Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:497 / 505
页数:9
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