Construction of engineered yeast strains for achieving better yields of biomass degradation

• Grant number: 18/02227-6
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: April 01, 2018
• End date: September 30, 2018
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Rafael Silva Rocha
• Grantee: Luísa Czamanski Nora
• Supervisor: Aindrila Mukhopadhyay
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: Lawrence Berkeley National Laboratory, United States
• Associated to the scholarship: 16/03763-3 - Unveiling the mechanisms of gene regulation at the single cell level in Trichoderma reesei, BP.MS

Abstract

Plant biomass degradation has been studied for years due to the incredible amount of byproducts generated during this process - ranging from biofuels to other noteworthy chemicals. The candidate current master's project is dedicated on building modular vectors based on synthetic biology for the transformation of fungi, more specifically T. reesei. The interest in this fungus arose because of its capability to produce high yields of biomass degrading enzymes, and the focus is in studying the expression of its cellulases promoters while degrading sugarcane bagasse, more specifically the ones from cel6a and cel7a genes. While in this continuous pursuit of better yields for biomass degradation, the idea of using a consortium of engineered microorganisms emerged. Since filamentous fungi are very difficult to be genetically manipulated and very sensible to the bioreactor environment, using R. toruloides, a newly emerging organism that is a good fermenter and that can degrade not only hemicellulose but lignin as well, seems to be an interesting solution. Mastering the manipulation of yeast genomes, one can create a community of several different strains of the same organism, each producing different genes, such as the enzymes of interest and/or resistance genes. Therefore, the main goals of this project are: (i) to learn how to use the CRISPR/Cas tools developed by Dr. Aindrila Mukhopadhyay's lab to manipulate yeast genomes; (ii) use these tools to create different yeast strains that produce the enzymes of interest, originally from T. reesei, in addition to acidic resistance genes. Our ultimate goal is obtaining the mutant strains and establishing the methodology, thus, we can bring it to Brazil to apply in our very own and abundant biomass: sugarcane bagasse. Additionally, all the technology and expertise regarding yeast manipulation will generate significant advances in metabolic modeling and synthetic biology in our country, as first steps in the aspiration of, one day, creating a community of yeast strains that can resist to harsh environments and produce high yields of lignocellulose-degrading enzymes. (AU)

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)

NORA, LUISA CZAMANSKI; WEHRS, MAREN; KIM, JOONHOON; CHENG, JAN-FANG; TARVER, ANGELA; SIMMONS, BLAKE A.; MAGNUSON, JON; HARMON-SMITH, MIRANDA; SILVA-ROCHA, RAFAEL; GLADDEN, JOHN M.; et al. A toolset of constitutive promoters for metabolic engineering of Rhodosporidium toruloides. Microbial Cell Factories, v. 18, JUN 29 2019. (12/22921-8, 18/02227-6)