Engineering a Balanced Acetyl Coenzyme A Metabolism in Saccharomyces cerevisiae for Lycopene Production through Rational and Evolutionary Engineering

Saccharomyces cerevisiaeis increasingly being used for the production of chemicals derived from acetyl coenzyme A(acetyl-CoA). However, the inadequate supply of cytosolic acetyl-CoA often leads to low yields. Here, we developed a novel strategyfor balancing acetyl-CoA metabolism and increasing the amount of the downstream product. First, the combination of acetaldehydedehydrogenase (eutE) and acetoacetyl-CoA thiolase (AtoB) was optimized to redirect the acetyl-CoAflux toward the targetpathway, with a 21-fold improvement in mevalonic acid production. Second, pathway engineering and evolutionary engineering wereconducted to attenuate the growth deficiency, and a 10-fold improvement of the maximum productivity was achieved. Third, acetyl-CoA carboxylase (ACC1) was dynamically downregulated as the complementary acetyl-CoA pathway, and the yield was improvedmore than twofold. Fourth, the most efficient and complementary acetyl-CoA pathways were combined, and thefinal strainproduced 68 mg/g CDW lycopene, which was among the highest yields reported inS. cerevisiae. This study demonstrates a newmethod of producing lycopene products by regulating acetyl-CoA metabolism