Extracellular vesicles carry cellulases in the industrial fungus Trichoderma reesei

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: BIOTECHNOLOGY FOR BIOFUELS; v. 12, JUN 15 2019.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 17/04206-3 - Elucidação das vias de sinalização celular mediadas por proteína quinase A (PKA), MAPK e cálcio durante expressão e secreção de celulases pelo fungo Trichoderma reesei
• Beneficiário: Roberto do Nascimento Silva
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 16/23233-9 - Caracterização da via de sinalização dependente de cálcio na regulação da expressão e secreção de celulases pelo fungo Trichoderma reesei (Hypocrea jecorina)
• Beneficiário: Renato Graciano de Paula
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado