Quantitative iTRAQ secretome analysis of Aspergillus niger reveals novel hydrolytic enzymes.
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.
Adav
Sunil S
SS
School of Biological Sciences, Nanyang Technological Universiy, 60 Nanyang Drive, Singapore.
Li
An A
AA
Manavalan
Arulmani
A
Punt
Peter
P
Sze
Siu Kwan
SK
D001234:Aspergillus niger / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D004798:Enzymes / Q000032:analysis* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008031:Lignin / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
D004798:Enzymes; C036909:lignocellulose; D008031:Lignin