卟啉生物合成。固定化酶。IV。对附着于琼脂糖的氨基乙酰丙酸脱水酶的研究。

Porphyrin biosynthesis. Immobilized enzymes. IV. Studies on aminolaevulate dehydratase attached to Sepharose.

作者信息

Stella A M, Wider de Xifra E, del C Batlle A M

出版信息

Mol Cell Biochem. 1977 Jul 5;16(2):97-104. doi: 10.1007/BF01732049.

DOI:10.1007/BF01732049

Abstract

Bovine liver aminolaevulate dehydratase (ALA-D) has been chemically attached to Sepharose 4B and its properties have been studied. The optimal conditions for coupling have been determined. It was found that the immobilized enzyme retained a significant percentage of the activity of the free enzyme. The coupling yield was rather high. The insolubilized enzyme requires both anaerobiosis and a thiol activator for maximal activity. It can be stored at 4 degrees C for long periods with little loss of activity and it can be repeatedly used without alteration of its enzymic capacity. Attachment of ALA-D to the gel has led to an enhanced thermal stability. pH optima of free and bound enzyme was the same while a small decrease in the Km of the matrix bonded ALA-D as compared to that of the soluble enzyme was observed. The use of the fixed-ALA-D for the preparation of PBG is described.

摘要

牛肝氨基乙酰丙酸脱水酶（ALA-D）已被化学连接到琼脂糖4B上，并对其性质进行了研究。已确定了偶联的最佳条件。发现固定化酶保留了游离酶相当大比例的活性。偶联产率相当高。不溶性酶需要厌氧环境和硫醇激活剂才能达到最大活性。它可以在4℃下长期保存，活性损失很小，并且可以重复使用而不改变其酶活性。ALA-D与凝胶的连接导致热稳定性增强。游离酶和结合酶的最适pH相同，而与可溶性酶相比，基质结合的ALA-D的Km略有降低。描述了使用固定化ALA-D制备胆色素原（PBG）的方法。