Purification and characterization of a type B feruloyl esterase (StFAE-A) from the thermophilic fungus Sporotrichum thermophile

Topakas, E; Stamatis, H; Biely, P; Christakopoulos, P

dc.contributor.author	Topakas, E	en
dc.contributor.author	Stamatis, H	en
dc.contributor.author	Biely, P	en
dc.contributor.author	Christakopoulos, P	en
dc.date.accessioned	2014-03-01T01:21:16Z
dc.date.available	2014-03-01T01:21:16Z
dc.date.issued	2004	en
dc.identifier.issn	0175-7598	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/16173
dc.subject	Cation Exchange	en
dc.subject	Enzyme	en
dc.subject	Enzyme Activity	en
dc.subject	ferulic acid	en
dc.subject	Phenolic Acid	en
dc.subject	Molecular Mass	en
dc.subject	Wheat Bran	en
dc.subject.classification	Biotechnology & Applied Microbiology	en
dc.subject.other	4 nitrophenyl 4 o feruloyl alpha arabinofuranoside	en
dc.subject.other	4 nitrophenyl 5 o feruloyl alpha arabinofuranoside	en
dc.subject.other	arabinofuranose	en
dc.subject.other	benzene derivative	en
dc.subject.other	butanol	en
dc.subject.other	caffeic acid methyl ester	en
dc.subject.other	esterase	en
dc.subject.other	ether derivative	en
dc.subject.other	ferulic acid	en
dc.subject.other	ferulic acid methyl ester	en
dc.subject.other	feruloyl esterase	en
dc.subject.other	fungal enzyme	en
dc.subject.other	hexane	en
dc.subject.other	methyl 4 coumarate	en
dc.subject.other	unclassified drug	en
dc.subject.other	water	en
dc.subject.other	xylan endo 1,3 beta xylosidase	en
dc.subject.other	enzyme	en
dc.subject.other	fungus	en
dc.subject.other	article	en
dc.subject.other	chromatofocusing	en
dc.subject.other	controlled study	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme analysis	en
dc.subject.other	enzyme purification	en
dc.subject.other	incubation time	en
dc.subject.other	ion exchange chromatography	en
dc.subject.other	molecular weight	en
dc.subject.other	nonhuman	en
dc.subject.other	pH	en
dc.subject.other	polyacrylamide gel electrophoresis	en
dc.subject.other	protein folding	en
dc.subject.other	Sporothrix	en
dc.subject.other	sporotrichum thermophile	en
dc.subject.other	wheat bran	en
dc.subject.other	1-Butanol	en
dc.subject.other	Caffeic Acids	en
dc.subject.other	Carboxylic Ester Hydrolases	en
dc.subject.other	Chromatography, Gel	en
dc.subject.other	Chromatography, Ion Exchange	en
dc.subject.other	Coumaric Acids	en
dc.subject.other	Dietary Fiber	en
dc.subject.other	Dimerization	en
dc.subject.other	Enzyme Stability	en
dc.subject.other	Hexanes	en
dc.subject.other	Hydrogen-Ion Concentration	en
dc.subject.other	Hydrophobicity	en
dc.subject.other	Hydroxybenzoic Acids	en
dc.subject.other	Isoelectric Point	en
dc.subject.other	Molecular Weight	en
dc.subject.other	Protein Subunits	en
dc.subject.other	Sporothrix	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Temperature	en
dc.subject.other	Xylosidases	en
dc.subject.other	Bacteria (microorganisms)	en
dc.subject.other	Corynascus heterothallicus	en
dc.subject.other	Fungi	en
dc.subject.other	Sporothrix	en
dc.subject.other	Sporotrichum	en
dc.subject.other	Triticum aestivum	en
dc.title	Purification and characterization of a type B feruloyl esterase (StFAE-A) from the thermophilic fungus Sporotrichum thermophile	en
heal.type	journalArticle	en
heal.identifier.primary	10.1007/s00253-003-1481-6	en
heal.identifier.secondary	http://dx.doi.org/10.1007/s00253-003-1481-6	en
heal.language	English	en
heal.publicationDate	2004	en
heal.abstract	A feruloyl esterase (StFAE-A) produced by Sporotrichum thermophile was purified to homogeneity. The purified homogeneous preparation of native StFAE-A exhibited a molecular mass of 57.0+/-1.5 kDa, with a mass of 33+/-1 kDa on SDS-PAGE. The pI of the enzyme was estimated by cation-exchange chromatofocusing to be at pH 3.1. The enzyme activity was optimal at pH 6.0 and 55-60 degreesC. The purified esterase was stable at the pH range 5.0-7.0. The enzyme retained 70% of activity after 7 h at 50 degreesC and lost 50% of its activity after 45 min at 55 degreesC and after 12 min at 60 degreesC. Determination of k(cat)/K-m revealed that the enzyme hydrolyzed methyl p-coumarate 2.5- and 12-fold more efficiently than methyl caffeate and methyl ferulate, respectively. No activity on methyl sinapinate was detected. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose and it hydrolyzed 4-nitrophenyl 5-O-trans-feruloyl-alpha-L-arabinofuranoside (NPh-5-Fe-Araf) 2-fold more efficiently than NPh-2-Fe-Araf. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with xylanase from S. thermophile (a maximum of 34% total ferulic acid released after 1 h incubation). StFAE-A by itself could release FA, but at a level almost 47-fold lower than that obtained in the presence of xylanase. The potential of StFAE-A for the synthesis of various phenolic acid esters was tested using a ternary water-organic mixture consisting of n-hexane, 1-butanol and water as a reaction system.	en
heal.publisher	SPRINGER-VERLAG	en
heal.journalName	Applied Microbiology and Biotechnology	en
dc.identifier.doi	10.1007/s00253-003-1481-6	en
dc.identifier.isi	ISI:000189195300010	en
dc.identifier.volume	63	en
dc.identifier.issue	6	en
dc.identifier.spage	686	en
dc.identifier.epage	690	en