Purification and mode of action of an alkali-resistant endo-1,4-β- glucanase from Bacillus pumilus

Christakopoulos, P; Hatzinikolaou, DG; Fountoukidis, G; Kekos, D; Claeyssens, M; Macris, BJ

dc.contributor.author	Christakopoulos, P	en
dc.contributor.author	Hatzinikolaou, DG	en
dc.contributor.author	Fountoukidis, G	en
dc.contributor.author	Kekos, D	en
dc.contributor.author	Claeyssens, M	en
dc.contributor.author	Macris, BJ	en
dc.date.accessioned	2014-03-01T01:15:05Z
dc.date.available	2014-03-01T01:15:05Z
dc.date.issued	1999	en
dc.identifier.issn	0003-9861	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/13336
dc.subject	Alkali resistant endoglucanase	en
dc.subject	Bacillus pumilus	en
dc.subject	Mode of action	en
dc.subject	Purification	en
dc.subject.classification	Biochemistry & Molecular Biology	en
dc.subject.classification	Biophysics	en
dc.subject.other	bacterial enzyme	en
dc.subject.other	cellulase	en
dc.subject.other	article	en
dc.subject.other	Bacillus pumilus	en
dc.subject.other	column chromatography	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme analysis	en
dc.subject.other	enzyme mechanism	en
dc.subject.other	enzyme purification	en
dc.subject.other	enzyme stability	en
dc.subject.other	nonhuman	en
dc.subject.other	priority journal	en
dc.subject.other	Alkalies	en
dc.subject.other	Bacillus	en
dc.subject.other	Catalysis	en
dc.subject.other	Cellobiose	en
dc.subject.other	Cellulase	en
dc.subject.other	Chelating Agents	en
dc.subject.other	Chromatography, Gel	en
dc.subject.other	Chromatography, Ion Exchange	en
dc.subject.other	Electrophoresis, Polyacrylamide Gel	en
dc.subject.other	Enzyme Activation	en
dc.subject.other	Enzyme Stability	en
dc.subject.other	Glucose	en
dc.subject.other	Hydrogen-Ion Concentration	en
dc.subject.other	Metals	en
dc.subject.other	Molecular Weight	en
dc.subject.other	Polysaccharides	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Surface-Active Agents	en
dc.subject.other	Bacillus pumilus	en
dc.subject.other	Bacteria (microorganisms)	en
dc.subject.other	Pumilus	en
dc.subject.other	Triticum aestivum subsp. spelta	en
dc.subject.other	Zea mays	en
dc.title	Purification and mode of action of an alkali-resistant endo-1,4-β- glucanase from Bacillus pumilus	en
heal.type	journalArticle	en
heal.identifier.primary	10.1006/abbi.1999.1102	en
heal.identifier.secondary	http://dx.doi.org/10.1006/abbi.1999.1102	en
heal.language	English	en
heal.publicationDate	1999	en
heal.abstract	Alkaline endo-1,4-beta-D-glucanase was secreted by Bacillus pumilus grown in submerged culture on a combination of oat spelt xylan and corn starch as carbon sources. The enzyme was purified to homogeneity by Sephacryl S-200 and Q-Sepharose column chromatography. The protein corresponded to molecular mass and pi values of 67 kDa and 3.7, respectively. The enzyme was optimally active at pH 7.0-8.0 and 60 degrees C and retained 50% of its optimum activity at pH 12. The most notable characteristic of the endoglucanase was its high stability up to pH 12 for 20 h at 30 degrees C. The enzyme hydrolyzed carboxymethylcellulose (CMC) and cello-oligosaccharides but was inactive on cellobiose, cellotriose, Avicel, xylan, 4-nitrophenyl-beta-D-glucoside, 4-nitrophenyl-beta-D-cellobioside, and 4-nitrophenyl-beta-D-xyloside. Analysis of reaction mixtures by HPLC revealed that the enzyme produced almost exclusively cellotriose when acted on CMC and appeared to hydrolyze cello-oligosaccharides by successively releasing cellotriose. The use of 4-methylumbelliferyl cello-oligosaccharides and the determination of bond cleavage frequency revealed that the enzyme preferentially hydrolyzed the third glycosidic bond adjacent to the glycon. The enzyme mediated a decrease in the viscosity of CMC associated with a release of only small amounts of reducing sugar. The enzyme activity was not inhibited by metal ions, surfactants, and chelating agents used as components of laundry detergents. (C) 1999 Academic Press.	en
heal.publisher	ACADEMIC PRESS INC	en
heal.journalName	Archives of Biochemistry and Biophysics	en
dc.identifier.doi	10.1006/abbi.1999.1102	en
dc.identifier.isi	ISI:000079617500007	en
dc.identifier.volume	364	en
dc.identifier.issue	1	en
dc.identifier.spage	61	en
dc.identifier.epage	66	en