Analysis of the proteins secreted by Trichoderma harzianum P49P11 under carbon-limited conditions

Autor(es)

Gelain, Lucas ^{[1, 2]} ; Pabst, Martin ^[1] ; da Cruz Pradella, Jose Geraldo ^[3] ; da Costa, Aline Carvalho ^[2] ; van der Wielen, Luuk ^{[1, 4]} ; van Gulik, Walter M. ^[1]

Afiliação do(s) autor(es)

^[1] Delft Univ Technol, Dept Biotechnol, Van der Maasweg 9, NL-2629 HZ Delft - Netherlands

^[2] Univ Estadual Campinas, Dev Proc & Prod, Av Albert Einstein 500, Campinas - Brazil

^[3] Univ Fed Sao Paulo, Inst Sci & Technol, Av Cesare Mansueto Giulio Lattes 1201, Sao Jose Dos Campos - Brazil

^[4] Univ Limerick, Bernal Inst, Limerick V94 T9PX - Ireland

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF PROTEOMICS; v. 227, SEP 15 2020.
• Citações Web of Science: 2

Resumo

The wild type strain Trichoderma harzianum was able to synthesize enzymes that can catalyse the hydrolysis of p-nitrophenyl-beta-D-glucopyranoside (PNPGase) in glucose-limited chemostat cultures. Fructose/glucose and sucrose conditions provided low levels of PNPGase activity. To investigate whether under these conditions other enzymes were produced, a shotgun proteomics analysis of their supernatants was performed. The analysis has indicated that the different carbon sources used influenced the amounts of proteins secreted including 1,3-beta-glucanosyltransferase, alpha-1,2-mannosidase, alpha-galactosidase and glucan 1,3-beta-glucosidase. The analysis has also suggested the presence of beta-glucosidase, which could also be represented by PNPGase activity. Intracellular metabolites were quantified during PNPGase production for the condition using 20 g/L of glucose in the feed and differences were observed, indicating that intracellular glucose could be inhibiting PNPGase production. Significance: This work shows that sugars such as glucose, fructose/glucose and sucrose can be used as substrates for the continuous synthesis of different enzymes under carbon-limited conditions by Trichoderma harzianum. As far as we know, this is the first work about the continuous synthesis of enzymes under carbon-limited conditions suggesting that different easily assimilated carbon sources can be used to generate different enzymatic cocktails. Each enzyme or uncharacterized protein suggested by shotgun proteomics has the potential to become a promising product for biotechnological applications. (AU)

• Processo FAPESP: 14/22537-9 - Modelagem matemática da produção de enzimas pelo Trichoderma harzianum em fermentação submersa
• Beneficiário: Lucas Gelain
• Modalidade de apoio: Bolsas no Brasil - Doutorado