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

Christakopoulos, P; Kekos, D; Macris, BJ; Claeyssens, M; Bhat, MK

dc.contributor.author	Christakopoulos, P	en
dc.contributor.author	Kekos, D	en
dc.contributor.author	Macris, BJ	en
dc.contributor.author	Claeyssens, M	en
dc.contributor.author	Bhat, MK	en
dc.date.accessioned	2014-03-01T01:11:26Z
dc.date.available	2014-03-01T01:11:26Z
dc.date.issued	1995	en
dc.identifier.issn	0003-9861	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/11640
dc.subject	affinity purification	en
dc.subject	endo-1,4-β-D-glucanase	en
dc.subject	Fusarium oxysporum	en
dc.subject	mode of action	en
dc.subject.classification	Biochemistry & Molecular Biology	en
dc.subject.classification	Biophysics	en
dc.subject.other	4 nitrophenyl beta glucoside	en
dc.subject.other	4 nitrophenylxyloside	en
dc.subject.other	carboxymethylcellulose	en
dc.subject.other	cellobiose	en
dc.subject.other	cellopentaose	en
dc.subject.other	cellotetraose	en
dc.subject.other	cellotriose	en
dc.subject.other	cellulase	en
dc.subject.other	hymecromone	en
dc.subject.other	microcrystalline cellulose	en
dc.subject.other	oligosaccharide	en
dc.subject.other	unclassified drug	en
dc.subject.other	xylan	en
dc.subject.other	affinity chromatography	en
dc.subject.other	article	en
dc.subject.other	controlled study	en
dc.subject.other	culture medium	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme purification	en
dc.subject.other	enzyme substrate	en
dc.subject.other	fusarium oxysporum	en
dc.subject.other	gel filtration	en
dc.subject.other	molecular weight	en
dc.subject.other	nonhuman	en
dc.subject.other	polyacrylamide gel electrophoresis	en
dc.subject.other	priority journal	en
dc.subject.other	Carboxymethylcellulose	en
dc.subject.other	Cellulase	en
dc.subject.other	Cellulose	en
dc.subject.other	Enzyme Stability	en
dc.subject.other	Fusarium	en
dc.subject.other	Glycosyltransferases	en
dc.subject.other	Heat	en
dc.subject.other	Hydrogen-Ion Concentration	en
dc.subject.other	Hymecromone	en
dc.subject.other	Isoelectric Point	en
dc.subject.other	Molecular Weight	en
dc.subject.other	Oligosaccharides	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Support, Non-U.S. Gov't	en
dc.subject.other	Fusarium	en
dc.subject.other	Fusarium oxysporum	en
dc.title	Purification and characterization of a less randomly acting endo-1,4-β-D-glucanase from the culture filtrates of Fusarium oxysporum	en
heal.type	journalArticle	en
heal.identifier.primary	10.1006/abbi.1995.1057	en
heal.identifier.secondary	http://dx.doi.org/10.1006/abbi.1995.1057	en
heal.language	English	en
heal.publicationDate	1995	en
heal.abstract	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
heal.publisher	ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS	en
heal.journalName	Archives of Biochemistry and Biophysics	en
dc.identifier.doi	10.1006/abbi.1995.1057	en
dc.identifier.isi	ISI:A1995QJ93300057	en
dc.identifier.volume	316	en
dc.identifier.issue	1	en
dc.identifier.spage	428	en
dc.identifier.epage	433	en