Cloning, characterization and functional expression of an alkalitolerant type C feruloyl esterase from Fusarium oxysporum

Moukouli, M; Topakas, E; Christakopoulos, P

dc.contributor.author	Moukouli, M	en
dc.contributor.author	Topakas, E	en
dc.contributor.author	Christakopoulos, P	en
dc.date.accessioned	2014-03-01T01:28:02Z
dc.date.available	2014-03-01T01:28:02Z
dc.date.issued	2008	en
dc.identifier.issn	0175-7598	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/18677
dc.subject	Alkalitolerant	en
dc.subject	Expression	en
dc.subject	Feruloyl esterase	en
dc.subject	Fusarium oxysporum	en
dc.subject	Gene cloning	en
dc.subject	Pichia pastoris	en
dc.subject.classification	Biotechnology & Applied Microbiology	en
dc.subject.other	Alkalitolerant	en
dc.subject.other	Feruloyl esterase	en
dc.subject.other	Fusarium oxysporum	en
dc.subject.other	Pichia pastoris	en
dc.subject.other	Amino acids	en
dc.subject.other	Cloning	en
dc.subject.other	Enzyme activity	en
dc.subject.other	Hydrolysis	en
dc.subject.other	Molecular weight	en
dc.subject.other	Yeast	en
dc.subject.other	Gene expression	en
dc.subject.other	alcohol oxidase	en
dc.subject.other	alkyl group	en
dc.subject.other	benzene derivative	en
dc.subject.other	coumaric acid	en
dc.subject.other	ester derivative	en
dc.subject.other	esterase	en
dc.subject.other	ferulic acid	en
dc.subject.other	feruloyl esterase	en
dc.subject.other	fungal DNA	en
dc.subject.other	fungal enzyme	en
dc.subject.other	glycoside	en
dc.subject.other	mating hormone alpha factor	en
dc.subject.other	methyl group	en
dc.subject.other	nitro derivative	en
dc.subject.other	recombinant enzyme	en
dc.subject.other	serine proteinase	en
dc.subject.other	unclassified drug	en
dc.subject.other	xylan endo 1,3 beta xylosidase	en
dc.subject.other	clone	en
dc.subject.other	enzyme activity	en
dc.subject.other	fungus	en
dc.subject.other	gene expression	en
dc.subject.other	hydrolysis	en
dc.subject.other	tolerance	en
dc.subject.other	yeast	en
dc.subject.other	amino acid sequence	en
dc.subject.other	AOX1 gene	en
dc.subject.other	article	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme analysis	en
dc.subject.other	enzyme specificity	en
dc.subject.other	enzyme stability	en
dc.subject.other	enzyme substrate	en
dc.subject.other	Fusarium oxysporum	en
dc.subject.other	genetic conservation	en
dc.subject.other	hydrolysis	en
dc.subject.other	molecular cloning	en
dc.subject.other	molecular weight	en
dc.subject.other	nonhuman	en
dc.subject.other	paper industry	en
dc.subject.other	pH	en
dc.subject.other	Pichia pastoris	en
dc.subject.other	promoter region	en
dc.subject.other	protein expression	en
dc.subject.other	protein motif	en
dc.subject.other	pulp mill	en
dc.subject.other	Saccharomyces cerevisiae	en
dc.subject.other	signal transduction	en
dc.subject.other	transcription regulation	en
dc.subject.other	Trichoderma	en
dc.subject.other	trichoderma longibrachiatum	en
dc.subject.other	wheat bran	en
dc.subject.other	Carboxylic Ester Hydrolases	en
dc.subject.other	Cloning, Molecular	en
dc.subject.other	Coumaric Acids	en
dc.subject.other	Fusarium	en
dc.subject.other	Gene Expression Regulation, Fungal	en
dc.subject.other	Hydrogen-Ion Concentration	en
dc.subject.other	Industrial Microbiology	en
dc.subject.other	Molecular Weight	en
dc.subject.other	Recombinant Proteins	en
dc.subject.other	Saccharomyces cerevisiae	en
dc.subject.other	Substrate Specificity	en
dc.subject.other	Fusarium oxysporum	en
dc.subject.other	Pichia pastoris	en
dc.subject.other	Saccharomyces cerevisiae	en
dc.subject.other	Trichoderma longibrachiatum	en
dc.subject.other	Triticum aestivum	en
dc.title	Cloning, characterization and functional expression of an alkalitolerant type C feruloyl esterase from Fusarium oxysporum	en
heal.type	journalArticle	en
heal.identifier.primary	10.1007/s00253-008-1432-3	en
heal.identifier.secondary	http://dx.doi.org/10.1007/s00253-008-1432-3	en
heal.language	English	en
heal.publicationDate	2008	en
heal.abstract	A hypothetical protein FoFaeC-12213 of Fusarium oxysporum was found to have high amino acid sequence identity with known type C feruloyl esterases (FAEs) containing a 13-amino acid conserved region flanking the characteristic G-X-S-X-G motif of a serine esterase. The putative FAE from the genomic DNA was successfully cloned in frame with the Saccharomyces cerevisiae α-factor secretion signal under the transcriptional control of the alcohol oxidase (AOX1) promoter and integrated in Pichia pastoris X-33 to confirm that the enzyme exhibits FAE activity. The molecular weight (62 kDa) and p I (6.8) were in agreement with the theoretical calculated values indicating the correct processing of the secretion signal in P. pastoris. The recombinant FAE was purified to its homogeneity 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 is a type C FAE showing broad hydrolytic activity against the four methyl esters of hydroxycinnamic acids and strong preference for the hydrolysis of n-propyl ferulate. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with xylanase from Trichoderma longibrachiatum (a maximum of 67% total FA released after 1-h incubation). The esterase showed broad pH stability making it an important candidate for alkaline applications such as pulp treatment in the paper industry. © 2008 Springer-Verlag.	en
heal.publisher	Springer-Verlag	en
heal.journalName	Applied Microbiology and Biotechnology	en
dc.identifier.doi	10.1007/s00253-008-1432-3	en
dc.identifier.isi	ISI:000255253200010	en
dc.identifier.volume	79	en
dc.identifier.issue	2	en
dc.identifier.spage	245	en
dc.identifier.epage	254	en