Extracellular vesicles carry cellulases in the industrial fungus Trichoderma reesei

Author(s)

de Paula, Renato Graciano ^[1] ; Campos Antonieto, Amanda Cristina ^[1] ; Vieira Nogueira, Karoline Maria ^[1] ; Costa Ribeiro, Liliane Fraga ^[1] ; Rocha, Marina Campos ^[2] ; Malavazi, Iran ^[2] ; Almeida, Fausto ^[1] ; Silva, Roberto Nascimento ^[1]

Affiliation

^[1] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Biochem & Immunol, BR-14049900 Ribeirao Preto, SP - Brazil

^[2] Univ Fed Sao Carlos, Ctr Ciencias Biol & Saude, Dept Genet & Evolucao, Sao Paulo - Brazil

• Document type: Journal article
• Source: BIOTECHNOLOGY FOR BIOFUELS; v. 12, JUN 15 2019.
• Web of Science Citations: 0

Abstract

BackgroundTrichoderma reesei is the most important industrial producer of lignocellulolytic enzymes. These enzymes play an important role in biomass degradation leading to novel applications of this fungus in the biotechnology industry, specifically biofuel production. The secretory pathway of fungi is responsible for transporting proteins addressed to different cellular locations involving some cellular endomembrane systems. Although protein secretion is an extremely efficient process in T. reesei, the mechanisms underlying protein secretion have remained largely uncharacterized in this organism.ResultsHere, we report for the first time the isolation and characterization of T. reesei extracellular vesicles (EVs). Using proteomic analysis under cellulose culture condition, we have confidently identified 188 vesicular proteins belonging to different functional categories. Also, we characterized EVs production using transmission electron microscopy in combination with light scattering analysis. Biochemical assays revealed that T. reesei extracellular vesicles have an enrichment of filter paper (FPase) and -glucosidase activities in purified vesicles from 24, 72 and 96, and 72 and 96h, respectively. Furthermore, our results showed that there is a slight enrichment of small RNAs inside the vesicles after 96h and 120h, and presence of hsp proteins inside the vesicles purified from T. reesei grown in the presence of cellulose.ConclusionsThis work points to important insights into a better understanding of the cellular mechanisms underlying the regulation of cellulolytic enzyme secretion in this fungus. (AU)

• FAPESP's process: 17/04206-3 - Elucidation of cellular signaling pathways mediated by protein kinase A (PKA), MAPK and calcium during expression and secretion of cellulases by the fungus Trichoderma reesei
• Grantee: Roberto do Nascimento Silva
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/23233-9 - Characterization of calcium-dependent signaling pathway in the secretion and cellulase expression by Trichoderma reesei (Hypocrea jecorina
• Grantee: Renato Graciano de Paula
• Support Opportunities: Scholarships in Brazil - Post-Doctoral