title: Secretomic survey of Trichoderma harzianum grown on plant biomass substrates.
abstract: The present work aims at characterizing T. harzianum secretome when the fungus is grown in synthetic medium supplemented with one of the four substrates: glucose, cellulose, xylan, and sugarcane bagasse (SB). The characterization was done by enzymatic assays and proteomic analysis using 2-DE/MALDI-TOF and gel-free shotgun LC-MS/MS. The results showed that SB induced the highest cellulolytic and xylanolytic activities when compared with the other substrates, while remarkable differences in terms of number and distribution of protein spots in 2-DE gels were also observed among the samples. Additionally, treatment of the secretomes with PNGase F revealed that most spot trails in 2-DE gels corresponded to N-glycosylated proteoforms. The LC-MS/MS analysis of the samples identified 626 different protein groups, including carbohydrate-active enzymes and accessory, noncatalytic, and cell-wall-associated proteins. Although the SB-induced secretome displayed the highest cellulolytic and xylanolytic activities, it did not correspond to a higher proteome complexity because CM-cellulose-induced secretome was significantly more diverse. Among the identified proteins, 73% were exclusive to one condition, while only 5% were present in all samples. Therefore, this study disclosed the variation of T. harzianum secretome in response to different substrates and revealed the diversity of the fungus enzymatic toolbox.
authors.affiliation: Laboratory of Biochemistry and Protein Chemistry, Department of Cell Biology, University of Brasilia , Asa Norte, Brasília, 70910-900 DF, Brazil.
authors.forename: Diana Paola
authors.initials: DP
authors.lastname: Gómez-Mendoza
authors.forename: Magno
authors.initials: M
authors.lastname: Junqueira
authors.forename: Luis Henrique Ferreira
authors.initials: LH
authors.lastname: do Vale
authors.forename: Gilberto Barbosa
authors.initials: GB
authors.lastname: Domont
authors.forename: Edivaldo Ximenes
authors.initials: EX
authors.lastname: Ferreira Filho
authors.forename: Marcelo Valle de
authors.initials: MV
authors.lastname: Sousa
authors.forename: Carlos André Ornelas
authors.initials: CA
authors.lastname: Ricart
mesh_terms: D018533:Biomass; D002480:Cellulase; D002482:Cellulose; D005656:Fungal Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D010449:Peptide Mapping; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D014242:Trichoderma / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism* / Q000502:physiology
chemical_list: D005656:Fungal Proteins; D020543:Proteome; D002482:Cellulose; C027433:bagasse; D002480:Cellulase