RESOLUTION OF CLOSTRIDIUM-STERCORARIUM CELLULASE BY FAST PROTEIN LIQUID-CHROMATOGRAPHY (FPLC)

被引：26

作者：

BRONNENMEIER, K ^{[1
]}

STAUDENBAUER, WL ^{[1
]}

机构：

[1] TECH UNIV MUNICH, INST MICROBIOL, ARCISSTR 21, D-8000 MUNICH 2, GERMANY

来源：

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | 1988年 / 27卷 / 5-6期

关键词：

D O I：

10.1007/BF00451608

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A fast and efficient separation procedure for the analysis of the cellulase components of the thermophilic anaerobe Clostridium stercorarium was developed. Culture supernatants were concentrated without loss of cellulase activity by tangential flow ultrafiltration. Resolution of the cellulase system was achieved by fast protein liquid chromatography (FPLC) on a Mono Q anion exchange column. Enzyme fractions were assayed for hydrolysis of Avicel, carboxymethylcellulose (CMC), .beta.-nitrophenyl-.beta.-D-cellobioside, and p-nitrophenyl-.beta.-D-glucoside. Two Avicelases, two .beta.-cellobiosidases, and one .beta.-glucosidase were identified and characterized by SDS-polyacrylamide electrophoresis and isoelectric focusing. On the basis of their activities towards CMC, Avicelase I was classified as endo-.beta.-glucanase and Avicelase II as exo-.beta.-glucanase. Efficient hydrolysis of microcrystalline cellulose was shown to result from the combined action of both Avicelases.

引用

页码：432 / 436

页数：5

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