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Use of pantothenate as a metabolic switch increases the genetic stability of farnesene producing Saccharomyces cerevisiae
Affiliation:1. Fermentation Engineering, Bielefeld University, Universitätsstr. 25, Bielefeld 33615, Germany;2. Microbial Genomics and Biotechnology, Center for Biotechnology (CeBiTec), Bielefeld University, Universitätsstr. 27, Bielefeld 33615, Germany;3. Austrian Center of Industrial Biotechnology (acib), Petersgasse 14, Graz 8010, Austria
Abstract:We observed that removing pantothenate (vitamin B5), a precursor to co-enzyme A, from the growth medium of Saccharomyces cerevisiae engineered to produce β-farnesene reduced the strain׳s farnesene flux by 70%, but increased its viability, growth rate and biomass yield. Conversely, the growth rate and biomass yield of wild-type yeast were reduced. Cultivation in media lacking pantothenate eliminates the growth advantage of low-producing mutants, leading to improved production upon scale-up to lab-scale bioreactor testing. An omics investigation revealed that when exogenous pantothenate levels are limited, acyl-CoA metabolites decrease, β-oxidation decreases from unexpectedly high levels in the farnesene producer, and sterol and fatty acid synthesis likely limits the growth rate of the wild-type strain. Thus pantothenate supplementation can be utilized as a “metabolic switch” for tuning the synthesis rates of molecules relying on CoA intermediates and aid the economic scale-up of strains producing acyl-CoA derived molecules to manufacturing facilities.
Keywords:Genetic stability  Pantothenate  Metabolic switch  Farnesene  Media development  Yeast
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