Varin L, Barron D, Ibrahim R K
Anal Biochem. 1987 Feb 15;161(1):176-80. doi: 10.1016/0003-2697(87)90669-5.
A novel enzyme assay for flavonoid sulfotransferase is described. It makes use of tetrabutylammonium dihydrogen phosphate which forms a pair of ions with the flavonoid sulfate esters formed. This renders the sulfate ester soluble in organic solvents such as ethyl acetate, whereas the sulfate donor, 3'-phosphoadenosine-5'-phosphosulfate, remains in the aqueous reaction mixture. The procedure is simple, rapid, and reproducible. It eliminates the need for chromatographic separation of the reaction products, except when their identification is required, and is suitable for use in the purification and kinetic studies of sulfotransferases.
描述了一种用于黄酮类硫酸转移酶的新型酶测定法。它利用磷酸二氢四丁铵与形成的黄酮类硫酸酯形成一对离子。这使得硫酸酯可溶于有机溶剂如乙酸乙酯,而硫酸供体3'-磷酸腺苷-5'-磷酸硫酸则保留在水性反应混合物中。该方法简单、快速且可重复。除了需要鉴定反应产物时,它无需对反应产物进行色谱分离,适用于硫酸转移酶的纯化和动力学研究。