Genetic engineering of Aspergillus oryzae for the production of recombinant (animal-free) bovine albumin by precision fermentation

• Grant number: 24/03771-2
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: August 01, 2025
• End date: February 29, 2028
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: André Ricardo de Lima Damasio
• Grantee: Gabriela Souza Alves Picciarelli
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 22/05731-2 - BEYOND: establishing a fungal cell factory for recombinant protein production, AP.TEM

Abstract

Filamentous fungi of the genus Aspergillus spp. They are commonly used for the production of recombinant proteins, being organisms consolidated as cellular factories. One of the advantages is the robustness of the protein folding and secretion system, in addition to the ability to carry out post-translational modifications, consolidating these hosts for large-scale protein production. Among the Aspergillus genus, the Aspergillus oryzae species is used in the food industry due to its safety, in addition to being a fungus listed as a GRAS (Generally Recognized as Safe) organism by the U.S. Food and Drug Administration (FDA). Therefore, the versatility of this fungus for the production of recombinant proteins becomes an integrated solution for the growing demand for alternative proteins in the food industry. The initiative aims to not only meet the dietary requirements of the growing global population, but also address the environmental challenge associated with conventional meat production. Focusing on alternative proteins, obtained from non-animal sources, the project aims to construct a strain of A. oryzae, using genetic engineering techniques such as CRISPR/Cas9. This strain will be optimized for the large-scale production of recombinant bovine albumin (rAlbumin), one of the main alternative proteins used as a specific component of the culture medium for cell growth in cultured meat. The approach involves genetic engineering techniques, such as CRISPR/Cas9, transformation, validation of mutant strains, production, purification of albumin, optimization in a 5L bioreactor, collection of transcriptomic data, validation of gene expression by RT-qPCR and detailed characterization of rAlbumin in cell culture. (AU)