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Purification of an extracellular cellulose‐binding endoglucanase of Cellulomonas sp. ATCC 21399 by affinity chromatography on H 3 PO 4 ‐swollen cellulose
Author(s) -
Poulsen Otto M.,
Petersen Lars W.
Publication year - 1987
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260290702
Subject(s) - cellulase , cellulose , affinity chromatography , chemistry , enzyme , hydrolysis , chromatography , biochemistry , microcrystalline cellulose , affinity electrophoresis
A cellulose‐binding endoglucanase (endoglucanase A) of Cellulomonas sp. ATCC 21399 was purified to immunological homogeneity by affinity chromatography ob H 3 PO 4 ‐swollen cellulose. This method of purification turned out to be an easy and very gentle method for obtaining a high yield of cellulose‐binding endoglucanase. The purified enzyme was immunologically homogeneous but appeared heterogeneous when analyzed by denaturing polyacrylamide gel electrophoresis. In addition to the cellulose‐binding of endoglucanase A, the enzyme also had a strong affinity for Concanavaline A, indicating that the enzyme was glycosylated. Purified endoglucanase A showed an endo mode of action on carboxymethylcellulose. The enzyme could hydrolyze microcrystalline cellulose when acting alone, and the enzyme had a high specific activity on H 3 PO 4 ‐swollen cellulose.