Quantitative iTRAQ secretome analysis of aspergillus niger reveals novel hydrolytic enzymes
article
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.
Topics
Food and Chemical Risk AnalysisA. niger; lignocellulosebioenergy and biorefineryiTRAQsecretomealpha arabinofuranosidasealpha glucosidasealpha glucosidase caspartic proteinaseaspergillopepsinbeta mannosidasebiofuelcellulasecytochrome c oxidaseenzymeglucan 1,4 alpha glucosidaseglucan synthaseglucose oxidaseglycosidasehemicellulasehydrolaselignocellulosemembrane proteinpeptidaseperoxidaseproteinprotein translocating transporter proteinproteinasesecretometripeptidyl peptidaseunclassified druganimal cellanimal experimentarticleAspergillus nigerbiofuel productionbiomasscontrolled studyculture mediumenzyme activityenzyme releaseenzyme specificityenzyme synthesisfermentationhydrolysisiTRAQ proteomicsliquid chromatography tandem mass spectrometrymass spectrometrymutantnonhumanpHpriority journalprotein analysisproteomicsquantitative analysistandem mass spectrometry
TNO Identifier
409288
ISSN
15353893
Source
Journal of Proteome Research, 9(8), pp. 3932-3940.
Pages
3932-3940
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