草酸青霉自溶物中β-葡萄糖苷酶的纯化及性质

Purification and properties of a beta-glucosidase from Penicillium oxalicum autolysates.

作者信息

Copa-Patiño J L, Rodriguez J, Pérez-Leblic M I

机构信息

Departamento de Microbiología y Parasitología, Universidad Alcalá de Henares, Madrid, Spain.

出版信息

FEMS Microbiol Lett. 1990 Jan 15;55(1-2):191-6. doi: 10.1016/0378-1097(90)90193-t.

DOI:10.1016/0378-1097(90)90193-t

PMID:2109721

Abstract

A beta-glucosidase from the medium of an autolyzed culture of Penicillium oxalicum has been purified by tannic acid precipitation, sephacryl S-200, DEAE-Biogel, CM-Biogel and Mono Q successively. The purification process produced a homogeneous band in the SDS-PAGE that correspond to a Mr of 133,500. The enzyme had a pl of 4, and the active optima were found at pH 5.5 and 55 degrees C. The enzyme hydrolyzed different substrates showing maximum affinity against p-nitrophenyl-beta-D-glucoside with a Km value of 0.37 mM. The beta-glucosidase was inhibited by Glucono-D-lactone but not by glucose in the concentration range of 1 to 10 mM. The enzyme was adsorbed by Concanavalin-A-Sepharose.

摘要

来自草酸青霉自溶培养物培养基的一种β-葡萄糖苷酶先后通过单宁酸沉淀、Sephacryl S-200、DEAE-琼脂糖凝胶、CM-琼脂糖凝胶和Mono Q进行了纯化。纯化过程在SDS-PAGE中产生了一条对应于133,500 Mr的均一谱带。该酶的pI为4，最适活性在pH 5.5和55℃时被发现。该酶能水解不同底物，对对硝基苯基-β-D-葡萄糖苷表现出最大亲和力，Km值为0.37 mM。在1至10 mM的浓度范围内，β-葡萄糖苷酶被葡萄糖酸-D-内酯抑制，但不被葡萄糖抑制。该酶被伴刀豆球蛋白A-Sepharose吸附。