Expression, characterization and structural modelling of a feruloyl esterase from the thermophilic fungus Myceliophthora thermophila

Topakas, E; Moukouli, M; Dimarogona, M; Christakopoulos, P

dc.contributor.author	Topakas, E	en
dc.contributor.author	Moukouli, M	en
dc.contributor.author	Dimarogona, M	en
dc.contributor.author	Christakopoulos, P	en
dc.date.accessioned	2014-03-01T02:09:00Z
dc.date.available	2014-03-01T02:09:00Z
dc.date.issued	2012	en
dc.identifier.issn	01757598	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/29766
dc.subject	Ferulic acid esterase	en
dc.subject	Heterologous expression	en
dc.subject	Myceliophthora thermophila	en
dc.subject	Pichia pastoris	en
dc.subject	Structural modelling	en
dc.subject.other	Ferulic acid esterase	en
dc.subject.other	Heterologous expression	en
dc.subject.other	Myceliophthora thermophila	en
dc.subject.other	Pichia Pastoris	en
dc.subject.other	Structural modelling	en
dc.subject.other	Cloning	en
dc.subject.other	Enzyme activity	en
dc.subject.other	Esterification	en
dc.subject.other	Hydrolysis	en
dc.subject.other	Substrates	en
dc.subject.other	Sugars	en
dc.subject.other	Three dimensional	en
dc.subject.other	Yeast	en
dc.subject.other	Esters	en
dc.subject.other	esterase	en
dc.subject.other	ferulic acid esterase	en
dc.subject.other	fungal enzyme	en
dc.subject.other	genomic DNA	en
dc.subject.other	unclassified drug	en
dc.subject.other	enzyme activity	en
dc.subject.other	gene expression	en
dc.subject.other	genome	en
dc.subject.other	homogeneity	en
dc.subject.other	hydrolysis	en
dc.subject.other	mitochondrial DNA	en
dc.subject.other	substrate	en
dc.subject.other	three-dimensional modeling	en
dc.subject.other	yeast	en
dc.subject.other	article	en
dc.subject.other	cloning	en
dc.subject.other	DNA synthesis	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme analysis	en
dc.subject.other	enzyme purification	en
dc.subject.other	enzyme structure	en
dc.subject.other	enzyme synthesis	en
dc.subject.other	hydrolysis	en
dc.subject.other	myceliophthora thermophila	en
dc.subject.other	nonhuman	en
dc.subject.other	Pichia pastoris	en
dc.subject.other	protein expression	en
dc.subject.other	protein secondary structure	en
dc.subject.other	protein tertiary structure	en
dc.subject.other	Sporothrix	en
dc.subject.other	Amino Acid Sequence	en
dc.subject.other	Carboxylic Ester Hydrolases	en
dc.subject.other	Cloning, Molecular	en
dc.subject.other	Gene Expression	en
dc.subject.other	Models, Molecular	en
dc.subject.other	Molecular Sequence Data	en
dc.subject.other	Molecular Weight	en
dc.subject.other	Pichia	en
dc.subject.other	Protein Conformation	en
dc.subject.other	Recombinant Proteins	en
dc.subject.other	Sequence Homology, Amino Acid	en
dc.subject.other	Sordariales	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Corynascus heterothallicus	en
dc.subject.other	Fungi	en
dc.subject.other	Pichia pastoris	en
dc.subject.other	Trichoderma longibrachiatum	en
dc.subject.other	Triticum aestivum	en
dc.title	Expression, characterization and structural modelling of a feruloyl esterase from the thermophilic fungus Myceliophthora thermophila	en
heal.type	journalArticle	en
heal.identifier.primary	10.1007/s00253-011-3612-9	en
heal.identifier.secondary	http://dx.doi.org/10.1007/s00253-011-3612-9	en
heal.publicationDate	2012	en
heal.abstract	A ferulic acid esterase (FAE) from the thermophilic fungus Myceliophthora thermophila (synonym Sporotrichum thermophile), belonging to the carbohydrate esterase family 1 (CE-1), was functionally expressed in methylotrophic yeast Pichia pastoris. The putative FAE from the genomic DNA was successfully cloned in P. pastoris X-33 to confirm that the enzyme exhibits FAE activity. The recombinant FAE was purified to its homogeneity (39 kDa) and subsequently characterized using a series of model substrates including methyl esters of hydroxycinnamates, alkyl ferulates and monoferuloylated 4-nitrophenyl glycosides. The substrate specificity profiling reveals that the enzyme shows a preference for the hydrolysis of methyl caffeate and p-coumarate and a strong preference for the hydrolysis of n-butyl and iso-butyl ferulate. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose, whilst it was found capable of de-esterifying acetylated glucuronoxylans. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with an M3 xylanase from Trichoderma longibrachiatum (a maximum of 41% total FA released after 1 h incubation). Prediction of the secondary structure of MtFae1a was performed in the PSIPRED server whilst modelling the 3D structure was accomplished by the use of the HH 3D structure prediction server. © Springer-Verlag 2011.	en
heal.journalName	Applied Microbiology and Biotechnology	en
dc.identifier.doi	10.1007/s00253-011-3612-9	en
dc.identifier.volume	94	en
dc.identifier.issue	2	en
dc.identifier.spage	399	en
dc.identifier.epage	411	en