In situ UDP-glucose regeneration unravels diverse functions of plant secondary product glycosyltransferases

Kazuyoshi Terasaka; Yuki Mizutani; Akito Nagatsu; Hajime Mizukami

doi:10.1016/j.febslet.2012.10.045

In situ UDP-glucose regeneration unravels diverse functions of plant secondary product glycosyltransferases

FEBS Lett. 2012 Dec 14;586(24):4344-50. doi: 10.1016/j.febslet.2012.10.045. Epub 2012 Nov 15.

Authors

Kazuyoshi Terasaka¹, Yuki Mizutani, Akito Nagatsu, Hajime Mizukami

Affiliation

¹ Graduate School of Pharmaceutical Sciences, Nagoya City University, Japan.

PMID: 23159939
DOI: 10.1016/j.febslet.2012.10.045

Abstract

The catalytic function of plant secondary product glycosyltransferases (PSPGs) was investigated by coupling the activities of recombinant flavonoid glucosyltransferases having different regiospecificities with sucrose synthase from Arabidopsis thaliana. In the present system, UDP, a product inhibitor of PSPGs, was removed from the reaction mixture and used for regeneration of UDP-glucose by AtSUS1. The in situ UDP-glucose regeneration system not only enhanced the glucosylation efficiency but also unraveled the novel regioselectivity of PSPGs. The effect of the system was shown to be because of the removal of UDP from the reaction system and not because of the additional supply of UDP-glucose.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / enzymology*
Arabidopsis Proteins / metabolism*
Glucosyltransferases / metabolism*
Glycosyltransferases / metabolism*
Substrate Specificity
Uridine Diphosphate / metabolism
Uridine Diphosphate Glucose / metabolism*

Substances

Arabidopsis Proteins
Uridine Diphosphate
Glycosyltransferases
Glucosyltransferases
sucrose synthase
flavonol 3-O-glucosyltransferase
Uridine Diphosphate Glucose