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Genetic engineering of Saccharomyces cerevisiae for xylose and glucose co-fermentation and second-generation ethanol production

Grant number: 18/00888-5
Support type:Scholarships in Brazil - Master
Effective date (Start): August 01, 2018
Effective date (End): February 29, 2020
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Cooperation agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal researcher:Leandro Vieira dos Santos
Grantee:João Gabriel Ribeiro Bueno
Home Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovações (Brasil). Campinas , SP, Brazil


The world energy matrix is based on the use of fossil fuels such as coal, gas and oil. However, in the last decades the worldwide community has expressed intense concern about the consequences caused by the use of these compounds, such as the increase of greenhouse gases emission. The second-generation ethanol (2G) from lignocellulosic biomass appears with a great potential of alternative energy production from a cleaner and renewable matrix. One of the major challenges in implementing this technology is the development of a robust microorganism capable to convert sugars from lignocellulosic materials into ethanol. Recently, our group developed a modified S. cerevisiae strain with high capacity to ferment the five-carbon sugar xylose, present in abundance in the lignocellulosic biomass. However, a major technological bottleneck in the 2G fermentation is the extensive time to consume xylose compared to glucose. Xylose is only assimilated after the majority consumption of glucose by the microorganism, caused by a competition of the sugar molecules to the yeast transporters, which have a higher affinity for glucose. There are few studies that covers this issue and no efficient transporter has been developed to solve this technological challenge. The principle of this project is the development of a strain with efficient co-fermentation on xylose and glucose, reducing the total time of 2G fermentation. For strain development, we will use the CRISPR-Cas9 methodology to delete three genes that encode hexokinases genes in two xylose fermenter strains developed by our group. The strains will be submitted to an adaptive evolution process in order to promote and identify mutations in sequences of sugar transporters and other genetic features that increase the affinity and transport xylose even in the presence of glucose, allowing the co-fermentation and decreasing the time of fermentation process.

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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)
RIBEIRO BUENO, JOAO GABRIEL; BORELLI, GUILHERME; RIBEIRO CORREA, THAMY LIVIA; FIAMENGHI, MATEUS BERNABE; JOS, JULIANA; DE CARVALHO, MURILO; DE OLIVEIRA, LEANDRO CRISTANTE; PEREIRA, GONCALO A. G.; DOS SANTOS, LEANDRO VIEIRA. Novel xylose transporter Cs4130 expands the sugar uptake repertoire in recombinantSaccharomyces cerevisiaestrains at high xylose concentrations. BIOTECHNOLOGY FOR BIOFUELS, v. 13, n. 1, . (17/08519-6, 18/00888-5, 17/05078-9)
PALERMO, GISELE CRISTINA DE LIMA; COUTOUNE, NATALIA; BUENO, JOAO GABRIEL RIBEIRO; MACIEL, LUCAS FERREIRA; DOSSANTOS, LEANDRO VIEIRA. Exploring metal ion metabolisms to improve xylose fermentation in Saccharomyces cerevisiae. MICROBIAL BIOTECHNOLOGY, v. 14, n. 5, . (20/07918-7, 18/00888-5, 18/06254-8, 17/08519-6)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
BUENO, João Gabriel Ribeiro. Engenharia genética de Saccharomyces cerevisiae para cofermentação de xilose e glicose e produção de etanol de segunda geração. 2020. Master's Dissertation - Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia Campinas, SP.

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