Biochemical and catalytic properties of an endoxylanase purified from the culture filtrate of Thermomyces lanuginosus ATCC 46882

Bennett, NA; Ryan, J; Biely, P; Vrsanska, M; Kremnicky, L; Macris, BJ; Kekos, D; Christakopoulos, P; Katapodis, P; Claeyssens, M; Nerinckx, W; Ntauma, P; Bhat, MK

dc.contributor.author	Bennett, NA	en
dc.contributor.author	Ryan, J	en
dc.contributor.author	Biely, P	en
dc.contributor.author	Vrsanska, M	en
dc.contributor.author	Kremnicky, L	en
dc.contributor.author	Macris, BJ	en
dc.contributor.author	Kekos, D	en
dc.contributor.author	Christakopoulos, P	en
dc.contributor.author	Katapodis, P	en
dc.contributor.author	Claeyssens, M	en
dc.contributor.author	Nerinckx, W	en
dc.contributor.author	Ntauma, P	en
dc.contributor.author	Bhat, MK	en
dc.date.accessioned	2014-03-01T01:13:36Z
dc.date.available	2014-03-01T01:13:36Z
dc.date.issued	1998	en
dc.identifier.issn	0008-6215	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/12606
dc.subject	Endoxylanase	en
dc.subject	Family 11	en
dc.subject	T. lanuginosus	en
dc.subject.classification	Biochemistry & Molecular Biology	en
dc.subject.classification	Chemistry, Applied	en
dc.subject.classification	Chemistry, Organic	en
dc.subject.other	Catalysis	en
dc.subject.other	Column chromatography	en
dc.subject.other	Enzyme kinetics	en
dc.subject.other	Filtration	en
dc.subject.other	Oligomers	en
dc.subject.other	pH	en
dc.subject.other	Polysaccharides	en
dc.subject.other	Proteins	en
dc.subject.other	Thin layer chromatography	en
dc.subject.other	Catalytic properties	en
dc.subject.other	Culture filtrate	en
dc.subject.other	Endoxylanase	en
dc.subject.other	Thermomyces lanuginosus	en
dc.subject.other	Plant cell culture	en
dc.subject.other	endo 1,4 beta xylanase	en
dc.subject.other	unclassified drug	en
dc.subject.other	xylan	en
dc.subject.other	xylose	en
dc.subject.other	article	en
dc.subject.other	catalysis	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme analysis	en
dc.subject.other	enzyme purification	en
dc.subject.other	fungus	en
dc.subject.other	hydrolysis	en
dc.subject.other	molecular weight	en
dc.subject.other	nonhuman	en
dc.subject.other	ph	en
dc.subject.other	priority journal	en
dc.subject.other	structure analysis	en
dc.subject.other	thin layer chromatography	en
dc.subject.other	Disaccharides	en
dc.subject.other	Endo-1,4-beta Xylanases	en
dc.subject.other	Enzyme Stability	en
dc.subject.other	Fungal Proteins	en
dc.subject.other	Hydrogen-Ion Concentration	en
dc.subject.other	Isoelectric Point	en
dc.subject.other	Mitosporic Fungi	en
dc.subject.other	Oligosaccharides	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Temperature	en
dc.subject.other	Xylans	en
dc.subject.other	Xylose	en
dc.subject.other	Xylosidases	en
dc.subject.other	Fungi	en
dc.subject.other	Thermomyces lanuginosus	en
dc.title	Biochemical and catalytic properties of an endoxylanase purified from the culture filtrate of Thermomyces lanuginosus ATCC 46882	en
heal.type	journalArticle	en
heal.identifier.primary	10.1016/S0008-6215(97)10076-3	en
heal.identifier.secondary	http://dx.doi.org/10.1016/S0008-6215(97)10076-3	en
heal.language	English	en
heal.publicationDate	1998	en
heal.abstract	An endoxylanase (1,4-β-D-xylan xylanohydrolase, EC 3.2.1.8) from the culture filtrates of T. lanuginosus ATCC 46882 was purified to homogeneity by DEAE-Sepharose and Bio-Gel P-30 column chromatographies. The purified endoxylanase had a specific activity of 888.8 μmol min-1 mg-1 protein and accounted for approximately 30% of the total protein secreted by this fungus. The molecular mass of native (non-denatured) and denatured endoxylanase were 26.3 and 25.7 kD as, respectively. Endoxylanase had a pI of 3.7 and was optimally active between pH 6.0-6.5 and at 75°C. The enzyme showed > 50% of its original activity between pH 5.5-9.0 and at 85 °C. The pH and temperature stability studies revealed that this endoxylanase was almost completely stable between pH 5.0-9.0 and up to 60°C for 5h and at pH 10.0 up to 55°C for 5 h. Thin-layer chromatography (TLC) analysis showed that endoxylanase released mainly xylose (Xyl) and xylobiose (Xyl2) from beechwood 4-O-methyl-D-glucuronoxylan, O-acetyl-4-Omethyl-D-glucuronoxylan and rhodymenan (a β-(1→3)-β(1→4)-xylan). Also, the enzyme released an acidic xylo-oligosaccharide from 4-O-methyl-D-glucuronoxylan, and an isomeric xylotetraose and an isomeric xylopentaose from rhodymenan. The enzyme hydrolysed [1-3H]-xylo-oligosaccharides in an endofashion, but the hydrolysis of [1-3H]-xylotriose appeared to proceed via transglycosylation, since the xylobiose was the predominant product. Endoxylanase was not active on pNPX and pNPC at 40 and 100 mM for up to 6h, but showed some activity towards pNPX at 100 nM after 20-24 h. The results suggested that the endoxylanase from T. lanuginosus belongs to family 11.Microbial endoxylanases have attracted considerable research interest in recent years mainly due to their potential application in food, animal feed, paper and pulp industries. An endoxylanase (1,4,-β-D-xylan xylanohydrolase, EC 3.2.1.8) from the culture filtrates of T. lanuginosus ATCC 46882 was purified to homogeneity by DEAE-Sepharose and Bio-Gel P-30 column chromatographies. The purified endoxylanase had a specific activity of 888.8 μmol min-1 mg-1 protein and accounted for approximately 30% of the total protein secreted by this fungus. The molecular mass of native (non-denatured) and denatured endoxylanase were 26.3 and 25.7 kD as, respectively. Endoxylanase had a pI of 3.7 and was optimally active between pH 6.0-6.5 and at 75°C. The enzyme showed > 50% of its original activity between pH 5.5-9.0 and at 85°C. The pH and temperature stability studies revealed that this endoxylanase was almost completely stable between pH 5.0-9.0 and up to 60°C for 5 h and at pH 10.0 up to 55°C for 5 h. Thin-layer chromatography (TLC) analysis showed that endoxylanase released mainly xylose (Xyl) and xylobiose (Xyl2) from beechwood 4-O-methyl-D-glucuronoxylan, O-acetyl-4-O-methyl-D-glucuronoxylan and rhodymenan (a β-(1→3)-β(1→4)-xylan). Also, the enzyme released an acidic xylo-oligosaccharide from 4-O-methyl-D-glucuronoxylan, and an isomeric xylotetraose and an isomeric xylopentaose from rhodymenan. The enzyme hydrolysed [1-3H]-xylo-oligosaccharides in an endofashion, but the hydrolysis of [1-3H]-xylotriose appeared to proceed via transglycosylation, since the xylobiose was the predominant product. Endoxylanase was not active on pNPX and pNPC at 40 and 100 mM for up to 6h, but showed some activity towards pNPX at 100 mM after 20-24h. The results suggested that the endoxylanase from T. lanuginosus belongs to family 11.	en
heal.publisher	Elsevier Sci B.V., Amsterdam, Netherlands	en
heal.journalName	Carbohydrate Research	en
dc.identifier.doi	10.1016/S0008-6215(97)10076-3	en
dc.identifier.isi	ISI:000074018100012	en
dc.identifier.volume	306	en
dc.identifier.issue	3	en
dc.identifier.spage	445	en
dc.identifier.epage	455	en