固定化作为稳定黄孢原毛平革菌INA-12产生的木质素过氧化物酶的一种工具。

Immobilization as a tool for the stabilization of lignin peroxidase produced by Phanerochaete chrysosporium INA-12.

作者信息

Asther M, Meunier J C

机构信息

Laboratoire de Chimie Biologique, I.N.R.A.-C.B.A.I., Institut National Agronomique, Centre de Grignon, Thiverval-Grignon, France.

出版信息

Appl Biochem Biotechnol. 1993 Jan-Feb;38(1-2):57-67. doi: 10.1007/BF02916412.

DOI:10.1007/BF02916412

PMID:8346905

Abstract

Lignin peroxidase immobilization was achieved by covalent coupling on CNBr-Sepharose 4B. Protein immobilization yield was around 80%. For veratryl alcohol oxidation, in the presence of hydrogen peroxide, both soluble and bound enzymes exhibited the same pH profile with an optimum near 2.5. Catalytic parameters (kc and Km) were seriously affected by immobilization. On the other hand, immobilization provided a noticeable stabilization of the enzyme against acidic pH and high temperatures. A 15-20 increase in the half-inactivation times at pH 2.2 and 2.7, respectively, could be observed. Bound enzyme was also much more thermostable than soluble.

摘要

通过共价偶联将木质素过氧化物酶固定在溴化氰活化的琼脂糖4B上。蛋白质固定化产率约为80%。对于藜芦醇氧化，在过氧化氢存在下，可溶性酶和固定化酶在相同的pH曲线下表现出相似的活性，最适pH值接近2.5。固定化对催化参数（kc和Km）有严重影响。另一方面，固定化使酶在酸性pH和高温下具有显著的稳定性。在pH 2.2和2.7时，固定化酶的半失活时间分别增加了15%-20%。固定化酶的热稳定性也远高于可溶性酶。