Physiological and genetic engineering of cytosolic redox metabolism in Saccharomyces cerevisiae for improved glycerol production.

摘要: Abstract Previous metabolic engineering strategies for improving glycerol production by Saccharomyces cerevisiae were constrained to a maximum theoretical yield of 1 mol·(mol glucose)−1 due the introduction rigid carbon, ATP or redox stoichiometries. In present study, we sought circumvent these constraints (i) maintaining flexibility at fructose-1,6-bisphosphatase and triosephosphate isomerase, while (ii) eliminating reactions that compete with formation cytosolic NADH (iii) enabling oxidative catabolism within mitochondrial matrix. aerobic, glucose-grown batch cultures S. strain, in which pyruvate decarboxylases external dehydrogenases respiratory chain-linked glycerol-3-phosphate dehydrogenase deleted this purpose, produced 0.90 mol·(mol glucose)−1. aerobic glucose-limited chemostat cultures, was ca. 25% lower, suggesting involvement an alternative glucose-sensitive mechanism oxidation NADH. Nevertheless, vivo generation additional co-feeding formate increased on glucose 1.08 mol mol−1. To our knowledge, is highest reported cerevisiae.