Genetic mapping of a bioethanol yeast strain reveals new targets for hydroxymethylfurfural- and thermotolerance

Author(s)

Bezerra de Mello, Fellipe da Silveira ; Venega Coradini, Alessandro Luis ; Carazzolle, Marcelo Falsarella ; Maneira, Carla ; Furlan, Monique ; Guimaraes Pereira, Goncalo Amarante ; Teixeira, Gleidson Silva

• Document type: Journal article
• Source: MICROBIOLOGICAL RESEARCH; v. 263, p. 13-pg., 2022-08-02.

Abstract

Current technology that enables bioethanol production from agricultural biomass imposes harsh conditions for Saccharomyces cerevisiae's metabolism. In this work, the genetic architecture of industrial bioethanol yeast strain SA-1 was evaluated. SA-1 segregant FMY097 was previously described as highly aldehyde resistant and here also as thermotolerant: two important traits for the second-generation industry. A Quantitative Trait Loci (QTL) mapping of 5-hydroxymethylfurfural (HMF) -resistant segregants of hybrid FMY097/BY4742 disclosed a region in chromosome II bearing alleles with uncommon non-synonymous (NS) single nucleotide polymorphisms (SNPs) in FMY097: MIX23, PKC1, SEA4, and SRO77. Allele swap to susceptible laboratory strain BY4742 revealed that SEA4(FMY097) enhances robustness towards HMF, but the industrial fitness could not be fully recovered. The genetic network arising from the causative genes in the QTL window suggests that intracellular signaling TOR (Target of Rapamycin) and CWI (Cell Wall Integrity) pathways are regulators of this phenotype in FMY097. Because the QTL mapping did not result in one major allelic contribution to the evaluated trait, a background effect in FMY097's HMF resistance is expected. Quantification of NADPH - cofactor implied in endogenous aldehyde detoxification reactions - supports the former hypothesis, given its high availability in FMY097. Regarding thermotolerance, SEA4(FMY097) grants BY4742 ability to grow in temperatures as high as 38 degrees C in liquid, while allele PKC1(FMY097) allows growth up to 40 degrees C in solid medium. Both SEA4(FMY097) and PKC1(FMY097) encode rare NS SNPs, not found in other > 1013S. cerevisiae. Altogether, these findings point towards crucial membrane and stress mediators for yeast robustness. (AU)

• FAPESP's process: 16/12852-0 - Implementation of a tetrad dissection method for Saccharomyces cerevisiae based on flow cytometry
• Grantee: Monique Furlan
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 15/06677-8 - Metabolic engineering and identification of quantitative traits loci related to HMF tolerance of Saccharomyces cerevisiae strain
• Grantee: Fellipe da Silveira Bezerra de Mello
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 13/08293-7 - CCES - Center for Computational Engineering and Sciences
• Grantee: Munir Salomao Skaf
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 16/02506-7 - Development of high-throughput approaches for the study of quantitative traits loci related to robustness of industrial yeasts (Saccharomyces cerevisiae)
• Grantee: Gleidson Silva Teixeira
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Regular Program Grants
• FAPESP's process: 14/26719-4 - Quantitative trait loci mapping of traits related to robustness in industrial yeast Pedra-2 (Saccharomyces cerevisiae)
• Grantee: Alessandro Luis Venega Coradini
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 18/03403-2 - Matabolic engineering of Saccharomyces cerevisiae wild strains using the CRISPR/Cas9 system
• Grantee: Carla Maneira da Silva
• Support Opportunities: Scholarships in Brazil - Master