Resolution ofClostridium stercorariumcellulase by fast protein liquid chromatography (FPLC)

✍ Scribed by Karin Bronnenmeier; Walter L. Staudenbauer

• Book ID: 104778978
• Publisher: Springer
• Year: 1988
• Tongue: English
• Weight: 458 KB
• Volume: 27
• Category: Article
• ISSN: 1432-0614
• DOI: 10.1007/bf00451608

✦ Synopsis

A fast and efficient separation procedure for the analysis of the cellulase components of the thermophilic anaerobe Clostridium sterco- rariurn 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), fl-nitrophenyl-fl-D-cellobioside, and p-nitrophenyl-fl-D-glucoside. Two Avicelases, two flcellobiosidases, and one fl-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-fl-glucanase and Avicelase II as exo-fl-glucanase. Efficient hydrolysis of microcrystalline cellulose was shown to result from the combined action of both Avicelases.