Einsatz der hochleistungsflüssigkeitschromatographie zur bestimmung der uridine-5′-diphosphoglucose:phenol-β-D-glucosyltransferase-aktivität in vitro mit partiell gereinigtem enzym aus pflanzen von beta vulgaris ssp. Rapacea var. altissima döll

Abstract Employment of high-performance liquid chromatography for the determination of uridine-5′-diphosphoglucose:phenol-β- D -glucosyltransferase activity in vitro by use of partially purified enzyme from plants of Beta vulgaris ssp. rapacea var. altissima Doll Activity of uridine-5′-diphosphoglucose (UDPG):phenol-β- D -glucosyltransferase (E.C. 2.4.1.35) in protein extracts from leaves of young plants of sugar beet ( Beta vulgaris ssp. rapacea var. altissima Doll) is detected by a cell-free system with UDPG as glucose donor and the methylester of p -hydroxybenzoic acid as substrate. The analysis of products formed is carried out by high-performance liquid chromatography combined with UV-detection. Integration of peak areas yields an relative measure of the rate of glucoside formation. This measure is used immediately for comparative studies and determination of Michaelis constant ( K M ) values. The method described here is suitable for analysing enzyme reactions in vitro without employment of radioactive substrates.