PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR BETA-GLUCOSIDASE WITH TRANSGLYCOSYLATION AND EXO-GLUCOSIDASE ACTIVITIES FROM FUSARIUM-OXYSPORUM

CHRISTAKOPOULOS, P; GOODENOUGH, PW; KEKOS, D; MACRIS, BJ; CLAEYSSENS, M; BHAT, MK

dc.contributor.author	CHRISTAKOPOULOS, P	en
dc.contributor.author	GOODENOUGH, PW	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	2014-03-01T01:10:10Z
dc.date.available	2014-03-01T01:10:10Z
dc.date.issued	1994	en
dc.identifier.issn	0014-2956	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/11329
dc.subject	fusarium oxysporum	en
dc.subject.classification	Biochemistry & Molecular Biology	en
dc.subject.other	SPOROTRICHUM-THERMOPHILE	en
dc.subject.other	TRICHODERMA-KONINGII	en
dc.subject.other	CELLULASE ACTIVITY	en
dc.subject.other	ETHANOL	en
dc.subject.other	FERMENTATION	en
dc.subject.other	VANDERWALT	en
dc.subject.other	ENZYMES	en
dc.title	PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR BETA-GLUCOSIDASE WITH TRANSGLYCOSYLATION AND EXO-GLUCOSIDASE ACTIVITIES FROM FUSARIUM-OXYSPORUM	en
heal.type	journalArticle	en
heal.identifier.primary	10.1111/j.1432-1033.1994.00379.x	en
heal.identifier.secondary	http://dx.doi.org/10.1111/j.1432-1033.1994.00379.x	en
heal.language	English	en
heal.publicationDate	1994	en
heal.abstract	An extracellular beta-glucosidase from Fusarium oxysporum was purified to homogeneity by gel filtration and ion-exchange chromatographies. The enzyme, a monomeric protein of 110 kDa, was maximally active at pH 5.0-6.0 and at 60 degrees C. It hydrolysed 1-->4-linked aryl-beta-glucosides and 1-->4-linked, 1-->3-linked and 1-->6-linked beta-glucosides. The apparent K-m and k(cat) values for p-nitrophenyl beta-D-glucopyranoside (4-NpGlcp) and cellobiose were 0.093 (K-m), 1.07 mM (k(cat)) and 1802 (K-m), 461.5 min(-1) (k(cat)), respectively. Glucose and gluconolactone inhibited the enzyme competitively with K-i values of 2.05 mM and 3.03 mu M, respectively. Alcohols activated the enzyme; butanol showed maximum effect (2.2-fold at 0.5 M) while methanol increased the activity by 1.4-fold at 1 M. The enzyme catalysed the synthesis of methylglucosides, ethylglucoside and propylglucosides, as well as trisaccharides in the presence of different alcohols and disaccharides, respectively In addition, the enzyme hydrolysed the unsubstituted and methylumbelliferyl cello-oligosaccharides [MeUmb(Glc)(n)] but the rate of hydrolysis decreased with increasing chain length. Analysis of prod ucts released from MeUmb(Glc)(n) as a function of time revealed that the enzyme attacked these substrates in a stepwise manner and from both ends. Thus, beta-glucosidase from F. oxysporum, with the above interesting properties, could be of commercial interest.	en
heal.publisher	SPRINGER VERLAG	en
heal.journalName	EUROPEAN JOURNAL OF BIOCHEMISTRY	en
dc.identifier.doi	10.1111/j.1432-1033.1994.00379.x	en
dc.identifier.isi	ISI:A1994PF80500012	en
dc.identifier.volume	224	en
dc.identifier.issue	2	en
dc.identifier.spage	379	en
dc.identifier.epage	385	en