通过在吲哚琼脂糖柱上进行亲和层析纯化吲哚-3-烷烃α-羟化酶。

Purification of indolyl-3-alkane alpha-hydroxylase by affinity chromatography on indolyl-agarose columns.

作者信息

Schmer G, Roberts J

出版信息

Biochim Biophys Acta. 1978 Nov 10;527(1):264-71. doi: 10.1016/0005-2744(78)90276-0.

Abstract

Indolyl-3-alkane alpha-hydroxylase was isolated from soil isolate organism, Pseudomonas XA, by affinity chromatography on indolyl-agarose, using different indole derivatives (L-tryptophan, N-acetyl-L-tryptophan, indole-3-carboxaldehyde and 3-indole-acrylic acid). With the exception of N-acetyl-L-tryptophan-agarose, excellent yields were obtained. The affinity chromatography step caused a 15-fold increase in the specific activity of the enzyme. The purity of indolyl-3-alkane alpha-hydroxylase was comparable to the preparations obtained by conventional isolation techniques; however, it showed a 7- to 10-times higher overall yield. Affinity purified indolyl-3-alkane alpha-hydroxylase exhibited essentially one band in polyacrylamide gel electrophoresis and on isoelectric focusing.

摘要

通过在吲哚琼脂糖上进行亲和层析，使用不同的吲哚衍生物（L-色氨酸、N-乙酰-L-色氨酸、吲哚-3-甲醛和3-吲哚丙烯酸），从土壤分离菌株假单胞菌XA中分离出吲哚-3-烷烃α-羟化酶。除了N-乙酰-L-色氨酸琼脂糖外，均获得了优异的产量。亲和层析步骤使该酶的比活性提高了15倍。吲哚-3-烷烃α-羟化酶的纯度与通过传统分离技术获得的制剂相当；然而，其总产率高出7至10倍。亲和纯化的吲哚-3-烷烃α-羟化酶在聚丙烯酰胺凝胶电泳和等电聚焦中基本上呈现一条带。