Quantitative iTRAQ Secretome Analysis of Aspergillus niger Reveals Novel Hydrolytic Enzymes
Autor: | Arulmani Manavalan, Siu Kwan Sze, Peter Punt, Sunil S. Adav, An A. Li |
---|---|
Rok vydání: | 2010 |
Předmět: |
Proteomics
Quantitative proteomics Lignin Biochemistry Microbiology Tandem Mass Spectrometry Gene Expression Regulation Fungal Glycoside hydrolase Chromatography High Pressure Liquid chemistry.chemical_classification biology Hydrolysis Aspergillus niger Fungal genetics General Chemistry Hydrogen-Ion Concentration biology.organism_classification Enzymes Secretory protein Enzyme chemistry biology.protein Peroxidase |
Zdroj: | Journal of Proteome Research. 9:3932-3940 |
ISSN: | 1535-3907 1535-3893 |
Popis: | 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. |
Databáze: | OpenAIRE |
Externí odkaz: |