GENETIC ENGINEERING OF A FUNGAL PLATFORM TO SECRETE LIGNOCELLULOLYTIC ENZYMES FOR OLIGOSACCHARIDE PRODUCTION

• Grant number: 17/10083-1
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: September 01, 2017
• End date: June 24, 2019
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: André Ricardo de Lima Damasio
• Grantee: Fabiano Jares Contesini
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 15/50612-8 - An integrated approach to explore a novel paradigm for biofuel production from lignocellulosic feedstocks, AP.TEM
• Associated scholarship(s): 17/26370-0 - GENETIC ENGINEERING OF FUNGAL STRAINS USING CRISPR/Cas9 MULTIPLEX TO PRODUCE TAILOR-MADE CELLULOLYTIC COCKTAILS, BE.EP.PD

Abstract

Several compounds of interest have been obtained through the fermentation of monosaccharides from the degradation of plant biomass. Nevertheless, the use of oligosaccharides, instead of monosaccharides, could bring advantages because it can decrease the competition with contaminants in the fermentation process. This strategy is possible through engineering of Saccharomyces cerevisiae strains that express specific oligosaccharides transporters. Bacteria from the genus Geobacillus, for instance, have transporters of cellodextrins and xylo-oligosaccharides and could be used as models to engineer yeasts. However, to obtain these oligosaccharides it is necessary the use of enzymatic cocktails that promote the partial degradation of polysaccharides. In this context, Trichoderma and Aspergillus are excellent platforms for enzyme production. The main objective of this project is to genetically engineer a fungal strain to produce an enzymatic cocktail in order to obtain cellodextrins and xylo-oligosaccharides with degree of polymerization between 5 and 10, from the degradation of pre-treated sugarcane straw and eucalyptus residues. We will evaluate one strain of Trichoderma reesei and one of Aspergilllus niger. One of these strains will be selected and we will delete genes of cellobiohydrolases, ²-glucosidases and ²-xylosidases using the CRISPR/Cas9 system. We will apply a proteomics approach to select important target enzymes for the formation of oligosaccharides and posteriorly overexpress these enzymes. The cultivation conditions and degradation of plant biomass will be optimized for the favorable formation of oligosaccharides.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

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. Improvement of homologous GH10 xylanase production by deletion of genes with predicted function in the Aspergillus nidulans secretion pathway. MICROBIAL BIOTECHNOLOGY, v. 13, n. 4, MAR 2020. (12/20549-4, 14/15403-6, 17/10083-1, 13/24988-5, 16/16306-0, 17/26315-9)

RUBIO, MARCELO VENTURA; FANCHINI TERRASAN, CESAR RAFAEL; CONTESINI, FABIANO JARES; ZUBIETA, MARIANE PALUDETTI; GERHARDT, JAQUELINE ALINE; OLIVEIRA, LEANDRO CRISTANTE; DE SOUZA SCHMIDT GONCALVES, ANY ELISA; ALMEIDA, FAUSTO; SMITH, BRADLEY JOSEPH; MARTINS FERREIRA DE SOUZA, GUSTAVO HENRIQUE; et al. Redesigning N-glycosylation sites in a GH3 beta-xylosidase improves the enzymatic efficiency. BIOTECHNOLOGY FOR BIOFUELS, v. 12, n. 1, NOV 14 2019. (12/20549-4, 13/08293-7, 16/16306-0, 17/22669-0, 17/10083-1, 19/00098-7, 14/15403-6, 13/24988-5)

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. Improvement of homologous GH10 xylanase production by deletion of genes with predicted function in the Aspergillus nidulans secretion pathway. MICROBIAL BIOTECHNOLOGY, v. 13, n. 4, p. 9-pg., 2020-03-25. (17/10083-1, 12/20549-4, 14/15403-6, 16/16306-0, 17/26315-9, 13/24988-5)