Improvement of homologous GH10 xylanase production by deletion of genes with predicted function in the Aspergillus nidulans secretion pathway

Author(s)

Zubieta, Mariane P. ; Gerhardt, Jaqueline A. ; Rubio, Marcelo V. ; Terrasan, Cesar R. F. ; Persinoti, Gabriela F. ; Antoniel, Everton P. ; Contesini, Fabiano J. ; Prade, Rolf A. ; Damasio, Andre

• Document type: Journal article
• Source: MICROBIAL BIOTECHNOLOGY; v. 13, n. 4, p. 9-pg., 2020-03-25.

Abstract

Filamentous fungi are important cell factories for large-scale enzyme production. However, production levels are often low, and this limitation has stimulated research focusing on the manipulation of genes with predicted function in the protein secretory pathway. This pathway is the major route for the delivery of proteins to the cell exterior, and a positive relationship between the production of recombinant enzymes and the unfolded protein response (UPR) pathway has been observed. In this study, Aspergillus nidulans was exposed to UPR-inducing chemicals and differentially expressed genes were identified by RNA-seq. Twelve target genes were deleted in A. nidulans recombinant strains producing homologous and heterologous GH10 xylanases. The knockout of pbnA (glycosyltransferase), ydjA (Hsp40 co-chaperone), trxA (thioredoxin) and cypA (cyclophilin) improved the production of the homologous xylanase by 78, 171, 105 and 125% respectively. Interestingly, these deletions decreased the overall protein secretion, suggesting that the production of the homologous xylanase was specifically altered. However, the production of the heterologous xylanase and the secretion of total proteins were not altered by deleting the same genes. Considering the results, this approach demonstrated the possibility of rationally increase the production of a homologous enzyme, indicating that trxA, cypA, ydjA and pbnA are involved in protein production by A. nidulans. (AU)

• FAPESP's process: 17/10083-1 - GENETIC ENGINEERING OF A FUNGAL PLATFORM TO SECRETE LIGNOCELLULOLYTIC ENZYMES FOR OLIGOSACCHARIDE PRODUCTION
• Grantee: Fabiano Jares Contesini
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 12/20549-4 - Secretion of heterologous glycoproteins in Aspergillus: effect of glycosylation pattern in functional parameters of glycosyl hydrolases
• Grantee: André Ricardo de Lima Damasio
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Young Investigators Grants
• FAPESP's process: 14/15403-6 - Aspergillus nidulans as a model to manipulate unfolded protein response-related genes
• Grantee: Mariane Paludetti Zubieta
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/16306-0 - Functional and biological analyses of LPMO (Lytic Polysaccharide Monooxigenases) and accessory proteins (non-CAZymes) for degradation of sugarcane straw by filamentous fungi
• Grantee: César Rafael Fanchini Terrasan
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 17/26315-9 - Influence of transcription factors on the secretion of enzymes in Aspergillus nidulans
• Grantee: Everton Paschoal Antoniel
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 13/24988-5 - Secretion of heterologous glycoproteins in Aspergillus: effect of glycosylation pattern in functional parameters of glycosyl hydrolases
• Grantee: Marcelo Ventura Rubio
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)