Purification and characterization of a less randomly acting endo-1,4-β-D-glucanase from the culture filtrates of Fusarium oxysporum (EN)

Christakopoulos, P (EN)

Bhat, MK (EN)

Macris, BJ (EN)

Claeyssens, M (EN)

Kekos, D (EN)

An extracellular endo-1,4-beta-D-glucanase from Fusarium oxysporum was purified by affinity chromatography and gel filtration. The enzyme purified in this way was homogeneous when judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing-polyacrylamide gel electrophoresis. The protein corresponded to a molecular mass and pI value of 41.7 kDa and 6.4, respectively. It was optimally active at pH 4.5 and at 55 degrees C. The enzyme hydrolyzed carboxymethylcellulose (CMC) and unsubstituted and substituted cello-oligosaccharides but was inactive on Avicel, filter paper, xylan, cellobiose, p-nitrophenyl-beta-D-glucoside, and p-nitrophenyl-beta-D-xyloside. However, the enzyme effected only a small change in viscosity of CMC per unit increase of reducing sugar. When cellotriose, cellotetraose, and cellopentaose were used as substrates, the enzyme released mainly cellobiose. Use of 4-methylumbelliferyl cello-oligosaccharides and the determination of bond cleavage frequency revealed that the enzyme preferentially hydrolyzed the glycosidic bond adjacent to 4-methylumbelliferone. Thus, the purified enzyme appeared to be a less randomly acting endoglucanase. (C) 1995 Academic Press, Inc. (EN)

journalArticle (EN)

unclassified drug (EN)

affinity chromatography (EN)

Molecular Weight (EN)

mode of action (EN)

cellotriose (EN)

cellulase (EN)

culture medium (EN)

fusarium oxysporum (EN)

Cellulose (EN)

Glycosyltransferases (EN)

Substrate Specificity (EN)

hymecromone (EN)

cellopentaose (EN)

Biochemistry & Molecular Biology (EN)

oligosaccharide (EN)

gel filtration (EN)

enzyme purification (EN)

cellotetraose (EN)

enzyme activity (EN)

Isoelectric Point (EN)

4 nitrophenyl beta glucoside (EN)

Fusarium (EN)

polyacrylamide gel electrophoresis (EN)

priority journal (EN)

Carboxymethylcellulose (EN)

Support, Non-U.S. Gov't (EN)

4 nitrophenylxyloside (EN)

Fusarium oxysporum (EN)

endo-1,4-β-D-glucanase (EN)

Enzyme Stability (EN)

microcrystalline cellulose (EN)

molecular weight (EN)

affinity purification (EN)

Biophysics (EN)

cellobiose (EN)

xylan (EN)

Oligosaccharides (EN)

article (EN)

Heat (EN)

Hymecromone (EN)

controlled study (EN)

carboxymethylcellulose (EN)

Cellulase (EN)

nonhuman (EN)

enzyme substrate (EN)

Hydrogen-Ion Concentration (EN)

Εθνικό Μετσόβιο Πολυτεχνείο

Ψηφιακό Αποθετήριο της Κεντρικής Βιβλιοθήκης του ΕΜΠ | Dspace@NTUA

Κεντρική Βιβλιοθήκη Ε.Μ.Π.

Ιδρυματικό Αποθετήριο

Δημοσιεύσεις μελών Δ.Ε.Π. σε περιοδικά

Archives of Biochemistry and Biophysics (EN)

Αγγλική γλώσσα

1995 (EN)

http://hdl.handle.net/123456789/11640

0003-9861 (EN)

1 (EN)

433 (EN)

316 (EN)

428 (EN)

10.1006/abbi.1995.1057 (EN)

ISI:A1995QJ93300057 (EN)

ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS (EN)