Highly parallelized laboratory evolution of wine yeasts for enhanced metabolic phenotypes

Adaptive Laboratory Evolution (ALE) of microorganisms can improve the efficiency of sustainable industrial processes important to the global economy. However, stochasticity and genetic background effects often lead to suboptimal outcomes during laboratory evolution. Here we report an ALE platform to circumvent these shortcomings through parallelized clonal evolution at an unprecedented scale. Using this platform, we evolved 10 4 yeast populations in parallel from many strains for eight desired wine fermentation-related traits. Expansions of both ALE replicates and lineage numbers broadened the evolutionary search spectrum leading to improved wine yeasts unencumbered by unwanted side effects. At the genomic level, evolutionary gains in metabolic characteristics often coincided with distinct chromosome amplifications and the emergence of side-effect syndromes that were characteristic of each selection niche. Several high-performing ALE strains exhibited desired wine fermentation kinetics when tested in larger liquid cultures, supporting their suitability for application. More broadly, our high-throughput ALE platform opens opportunities for rapid optimization of microbes which otherwise could take many years to accomplish. Synopsis Evolution of ten thousand yeast populations from many strains for eight desired wine fermentation related traits broadened the evolutionary search spectrum and lead to wine yeasts with improved metabolic characteristics. Expansions of both replicates and lineage numbers led to better performing wine yeasts that were unencumbered by unwanted side-effects. Evolutionary gains coincided with distinct side effect syndromes that were characteristic of each selection niche. High-performing strains exhibited desired wine fermentation kinetics when tested in larger liquid cultures. Evolution of ten thousand yeast populations from many strains for eight desired wine fermentation related traits broadened the evolutionary search spectrum and lead to wine yeasts with improved metabolic characteristics.