Evaluation of Saccharomyces cerevisiae modified via CRISPR/Cas9 as a cellulosic platform microorganism in simultaneously saccharification and fermentation processes

Author(s)

de Melo, Allan H. F. ; Nunes, Alexia L. ; Carvalho, Priscila H. ; da Silva, Marcos F. ; Teixeira, Gleidson S. ; Goldbeck, Rosana

• Document type: Journal article
• Source: Bioprocess and Biosystems Engineering; v. N/A, p. 9-pg., 2022-08-06.

Abstract

The nonrenewable character and deleterious effects of fossil fuels foster the need for cleaner and more inexhaustible energy sources, such as bioethanol. Especially from lignocellulosic biomasses. However, the economic viability of this product in the market depends on process optimization and cost reduction. This research applied a sequential experimental project to investigate the process of enzymatic saccharification and simultaneous fermentation to produce ethanol with sugarcane bagasse. The differential of the work was the application of the strain of Saccharomyces cerevisiae AGY001 which was improved by evolutionary engineering to become thermotolerant and by a heterologous expression based on genomic integration by CRISPR/Cas9 to produce endoglucanase and beta-glucosidase (AsENDO-AsBGL). The maximum ethanol yield found was 89% of the maximum theoretical yield (released sugars), obtained at temperature concentrations, sugarcane bagasse and inoculum at 40 degrees C, 16.5%, and 4.0 g/L, respectively (12.5 FPU/g bagasse). The mathematical model obtained can predict approximately 83% of the data set with 95% confidence. Therefore, these findings demonstrated the potential of sugarcane bagasse and S. cerevisiae AGY001 strain (CRISPR/Cas9 modified) in bioethanol production without the need for impractical selection media on an industrial scale, in addition to providing useful insights for the development of SSF processes. (AU)

• FAPESP's process: 16/04602-3 - Development of Saccharomyces cerevisiae strains through heterologous expression of cellulases and evolutionary engineering aiming for simultaneous saccharification and fermentation of lignocellulosic biomass
• Grantee: Rosana Goldbeck Coelho
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Regular Program Grants
• FAPESP's process: 15/20630-4 - Biorefinery development integrated to a bioethanol sugar cane plant with zero CO2 emission: routes to convert renewable resources to bio-products and bio-electricity
• Grantee: Rubens Maciel Filho
• Support Opportunities: Research Projects - Thematic Grants