Beneficial mutations for carotenoid production identified from laboratory-evolved Saccharomyces cerevisiae

Author:

Godara Avinash1,Rodriguez Maria Alejandra Gomez1,Weatherston Joshua D1,Peabody George L2,Wu Hung-Jen1,Kao Katy C13

Affiliation:

1. grid.264756.4 0000 0004 4687 2082 Department of Chemical Engineering Texas A&M University College Station TX USA

2. grid.135519.a 0000 0004 0446 2659 Oak Ridge National Laboratory 37831 Oak Ridge TN USA

3. grid.186587.5 0000 0001 0722 3678 Department of Chemical and Materials Engineering San Jose State University One Washington Square 95129 San Jose CA USA

Abstract

Abstract Adaptive laboratory evolution (ALE) is a powerful tool used to increase strain fitness in the presence of environmental stressors. If production and strain fitness can be coupled, ALE can be used to increase product formation. In earlier work, carotenoids hyperproducing mutants were obtained using an ALE strategy. Here, de novo mutations were identified in hyperproducers, and reconstructed mutants were explored to determine the exact impact of each mutation on production and tolerance. A single mutation in YMRCTy1-3 conferred increased carotenoid production, and when combined with other beneficial mutations led to further increased β-carotene production. Findings also suggest that the ALE strategy selected for mutations that confer increased carotenoid production as primary phenotype. Raman spectroscopy analysis and total lipid quantification revealed positive correlation between increased lipid content and increased β-carotene production. Finally, we demonstrated that the best combinations of mutations identified for β-carotene production were also beneficial for production of lycopene.

Funder

National Science Foundation

Publisher

Oxford University Press (OUP)

Subject

Applied Microbiology and Biotechnology,Biotechnology,Bioengineering

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