Fluorescence-based approach for the effective development of industrial Saccharomyces cerevisiae strains

Alessandro Luis Venega Coradini,Alessandro Dos Santos Farias,Gleidson Silva Teixeira,Goncalo Amarante Guimaraes Pereira,Monique Furlan

doi:10.20396/revpibic2620181060

Alessandro Luis Venega Coradini, Alessandro Dos Santos Farias + Show 3 more

Open Access

https://doi.org/10.20396/revpibic2620181060

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Abstract

The laboratory evolution based on sexual hybridization – haploids breeding – is a powerful approach to study and develop Saccharomyces cerevisiae strains applied to the production of renewable fuels and chemicals. In this context, we developed a simple and efficient method based on reporter proteins expression and fluorescence-activated cell sorting (FACS). Exploring the natural genetic variability of two industrial Brazilian strains we obtained haploids showing superior biomass production under tree industrially relevant stress condition. In conclusion, this approach enables the isolation of an unlimited number of recombinants haploids requiring minimal and transient engineering, providing a powerful tool for create new superior hybrids through breeding, map genes related to industrially relevant traits, improvement of commercial strains and development of new ones.

Highlights

In the context of developing more efficient yeasts for production of renewable fuels and chemicals, the laboratory sexual evolution is a powerful approach to explore genetic variability
The coding sequences for two fluorescent proteins were fused respectively to MATα-specific STE3 and MATaspecific STE2 promoters resulting in the pMF002
To confirm its functionality the pMF002 was transformed into industrial strain JAY270 (MATa/α) and its derived haploids JAY289 (MATa) and JAY290 (MATα)[4]

Summary

Introduction

In the context of developing more efficient yeasts for production of renewable fuels and chemicals, the laboratory sexual evolution is a powerful approach to explore genetic variability. As demonstrated by McDonald and colleagues[1] the sex speeds adaptation and alters its molecular signature by allowing natural selection to more efficiently sort beneficial from deleterious mutations. Yeasts naturally adapted to industrial environment – industrial strains – are considered appropriate platforms for the development of commercial strains[2], combine industrially relevant features by breeding require the isolation of a large number of recombinant haploids, typically obtained by laborious manual dissection of the tetrads.

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