通过亲和层析法从兔脑中简单快速地纯化色氨酸5-单加氧酶。

Simple and rapid purification of tryptophan 5-monooxygenase from rabbit brain by affinity chromatography.

作者信息

Nakata H, Fujisawa H

出版信息

J Biochem. 1981 Aug;90(2):567-9. doi: 10.1093/oxfordjournals.jbchem.a133508.

DOI:10.1093/oxfordjournals.jbchem.a133508

Abstract

A simple and rapid method for isolating tryptophan 5-monooxygenase [L-tryptophan, tetrahydropteridine:oxygen oxidoreductase (5-hydroxylating), EC 1.14.16.4] was reported. The method involves adsorption on calcium phosphate gel and affinity chromatography on agarose coupled with dimethyltetrahydropteridine. Tryptophan 5-monooxygenase was purified 1,100-fold from a rabbit brain extract to a specific activity of 15.9 nmol/min.mg, which is far higher than that of the highly purified preparation reported by Tong and Kaufman ((1975) J. Biol. Chem. 250, 4152-4158). It was also demonstrated that this method was applicable to the purification of rat brain tryptophan 5-monooxygenase.

摘要

报道了一种简单快速的分离色氨酸5-单加氧酶[L-色氨酸，四氢生物蝶呤：氧氧化还原酶（5-羟化），EC 1.14.16.4]的方法。该方法包括吸附于磷酸钙凝胶以及在与二甲基四氢生物蝶呤偶联的琼脂糖上进行亲和层析。色氨酸5-单加氧酶从兔脑提取物中纯化了1100倍，比活性达到15.9 nmol/分钟·毫克，这远远高于Tong和Kaufman（(1975)《生物化学杂志》250, 4152 - 4158）报道的高度纯化制剂。还证明了该方法适用于大鼠脑色氨酸5-单加氧酶的纯化。