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Quantitative iTRAQ secretome analysis of aspergillus niger reveals novel hydrolytic enzymes

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Author: Adav, S.S. · Li, A.A. · Manavalan, A. · Punt, P. · Sze, S.K.
Type:article
Date:2010
Institution: TNO Kwaliteit van Leven
Source:Journal of Proteome Research, 8, 9, 3932-3940
Identifier: 409288
Keywords: Biology · Food and Chemical Risk Analysis · A. niger; lignocellulose · bioenergy and biorefinery · iTRAQ · secretome · alpha arabinofuranosidase · alpha glucosidase · alpha glucosidase c · aspartic proteinase · aspergillopepsin · beta mannosidase · biofuel · cellulase · cytochrome c oxidase · enzyme · glucan 1,4 alpha glucosidase · glucan synthase · glucose oxidase · glycosidase · hemicellulase · hydrolase · lignocellulose · membrane protein · peptidase · peroxidase · protein · protein translocating transporter protein · proteinase · secretome · tripeptidyl peptidase · unclassified drug · animal cell · animal experiment · article · Aspergillus niger · biofuel production · biomass · controlled study · culture medium · enzyme activity · enzyme release · enzyme specificity · enzyme synthesis · fermentation · hydrolysis · iTRAQ proteomics · liquid chromatography tandem mass spectrometry · mass spectrometry · mutant · nonhuman · pH · priority journal · protein analysis · proteomics · quantitative analysis · tandem mass spectrometry

Abstract

The natural lifestyle of Aspergillus niger made them more effective secretors of hydrolytic proteins and becomes critical when this species were exploited as hosts for the commercial secretion of heterologous proteins. The protein secretion profile of A. niger and its mutant at different pH was explored using iTRAQ-based quantitative proteomics approach coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). This study characterized 102 highly confident unique proteins in the secretome with zero false discovery rate based on decoy strategy. The iTRAQ technique identified and relatively quantified many hydrolyzing enzymes such as cellulases, hemicellulases, glycoside hydrolases, proteases, peroxidases, and protein translocating transporter proteins during fermentation. The enzymes have potential application in lignocellulosic biomass hydrolysis for biofuel production, for example, the cellulolytic and hemicellulolytic enzymes glucan 1,4-alpha-glucosidase, alpha-glucosidase C, endoglucanase, alpha l-arabinofuranosidase, beta-mannosidase, glycosyl hydrolase; proteases such as tripeptidyl-peptidase, aspergillopepsin, and other enzymes including cytochrome c oxidase, cytochrome c oxidase, glucose oxidase were highly expressed in A. niger and its mutant secretion. In addition, specific enzyme production can be stimulated by controlling pH of the culture medium. Our results showed comprehensive unique secretory protein profile of A. niger, its regulation at different pH, and the potential application of iTRAQ-based quantitative proteomics for the microbial secretome analysis. © 2010 American Chemical Society.