Abstract
Ethyl crotonate is one of the most important spices for food and is also a chemical intermediate in various chemical essences. In this study, Saccharomyces cerevisiae AY14α was engineered to produce ethyl crotonate. Firstly, several expression cassettes with different strengths were constructed for driving functional genes to balance metabolic flow, the 3-hydroxybutyryl-CoA dehydrogenase (Hbd) and 3-hydroxybutyryl-CoA dehydratase (Crt) genes from Clostridium tyrobutyricum, and the alcohol acyltransferase (AAT) gene from Fragaria×ananassa were heterologously expressed in S. cerevisiae, the engineered strain Ct-HCS produced 26.01 ± 3.57 mg/L of ethyl crotonate. Secondly, the Hbd and Crt genes from different microorganism, and the AAT genes from different fruit were expressed for the biosynthetic pathway optimization, the recombinant strain Ck-HCVL accumulated 52.53 ± 1.10 mg/L of ethyl crotonate. Finally, the acetyl-CoA acetyltransferase and alcohol acyltransferase genes were overexpressed in strain Ck-HCVL, the engineered strain Ck-HCDVL-E produced 125.59 ± 2.04 mg/L of ethyl crotonate. The work described in this paper provided a reference for environmentally friendly and large-scale industrial production of ethyl crotonate from renewable resources.
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