Cloning and optimized expression of a GH-11 xylanase from Fusarium oxysporum in Pichia pastoris

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	2014-03-01T01:35:24Z
dc.date.available	2014-03-01T01:35:24Z
dc.date.issued	2011	en
dc.identifier.issn	1871-6784	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/21034
dc.subject	fusarium oxysporum	en
dc.subject	pichia pastoris	en
dc.subject.other	Alcohol oxidase promoters	en
dc.subject.other	Culture conditions	en
dc.subject.other	Enzymatic activities	en
dc.subject.other	Enzyme production	en
dc.subject.other	Family 11	en
dc.subject.other	Flask culture	en
dc.subject.other	Fusarium oxysporums	en
dc.subject.other	Glycosyl hydrolases	en
dc.subject.other	Initial pH	en
dc.subject.other	Methanol concentration	en
dc.subject.other	Methylotrophic yeasts	en
dc.subject.other	P. pastoris	en
dc.subject.other	Pichia Pastoris	en
dc.subject.other	Signal peptide	en
dc.subject.other	Xylanase activity	en
dc.subject.other	Xylanases	en
dc.subject.other	Cloning	en
dc.subject.other	Enzymes	en
dc.subject.other	Genes	en
dc.subject.other	Methanol	en
dc.subject.other	Yeast	en
dc.subject.other	alcohol oxidase	en
dc.subject.other	glycosidase	en
dc.subject.other	glycosyl hydrolase 11	en
dc.subject.other	methanol	en
dc.subject.other	unclassified drug	en
dc.subject.other	article	en
dc.subject.other	controlled study	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme synthesis	en
dc.subject.other	fungal genome	en
dc.subject.other	Fusarium oxysporum	en
dc.subject.other	molecular cloning	en
dc.subject.other	nonhuman	en
dc.subject.other	nucleotide sequence	en
dc.subject.other	pH	en
dc.subject.other	Pichia pastoris	en
dc.subject.other	plasmid	en
dc.subject.other	priority journal	en
dc.subject.other	protein expression	en
dc.subject.other	Cloning, Molecular	en
dc.subject.other	Endo-1,4-beta Xylanases	en
dc.subject.other	Fusarium	en
dc.subject.other	Pichia	en
dc.subject.other	Protein Engineering	en
dc.subject.other	Recombinant Proteins	en
dc.subject.other	Fusarium oxysporum	en
dc.subject.other	Pichia pastoris	en
dc.title	Cloning and optimized expression of a GH-11 xylanase from Fusarium oxysporum in Pichia pastoris	en
heal.type	journalArticle	en
heal.identifier.primary	10.1016/j.nbt.2011.03.002	en
heal.identifier.secondary	http://dx.doi.org/10.1016/j.nbt.2011.03.002	en
heal.language	English	en
heal.publicationDate	2011	en
heal.abstract	The endo-1,4-β-xylanase gene xyn11a from Fusarium oxysporum, member of the fungal glycosyl hydrolase (GH) family 11, was cloned and expressed in Pichia pastoris. The mature xylanase gene, which generates after the excision of one intron and the secreting signal peptide, was placed under the control of an alcohol oxidase promoter (AOX1) in the plasmid pPICZαC. The final construction was integrated into the genome of the methylotrophic yeast P. pastoris X33 and the ability to produce xylanase activity was evaluated in flask cultures. Recombinant P. pastoris efficiently secreted xylanase into the medium and produced high level of enzymatic activity (110. U/ml) after 216 hours of growth, under methanol induction. To achieve higher enzyme production, the influence of initial pH, methanol concentration, agitation and flask design was evaluated. Under optimum culture conditions, production of the recombinant xylanase increased by 50%, reaching a final yield of 170. U/ml, underpinning aeration as the most important factor in improving enzyme production. © 2011 Elsevier B.V.	en
heal.publisher	ELSEVIER SCIENCE BV	en
heal.journalName	New Biotechnology	en
dc.identifier.doi	10.1016/j.nbt.2011.03.002	en
dc.identifier.isi	ISI:000292717500011	en
dc.identifier.volume	28	en
dc.identifier.issue	4	en
dc.identifier.spage	369	en
dc.identifier.epage	374	en