Method for constructing a recombinant bacterium with high productivity of beta-elemene and germacrene a

Abstract

The present invention relates to a method for constructing a recombinant bacterium with high production of β-elemene and germacrene A. Firstly, β-elemene and germacrene A are synthesized from scratch through the screening of germacrene A synthase and the overexpression of the mevalonate pathway; then, the availability of acetyl-CoA, pyruvate, and glyceraldehyde-3-phosphate in the farnesyl diphosphate pathway is ensured by deleting competing pathways in the central carbon metabolism; next, the present invention uses lycopene color as a high-throughput screening method and obtains an optimized NSY305N through error-prone PCR. Finally, in shake flasks, strain β-EL-4 constructed through key pathway enzymes, efflux engineering, and translation engineering produced 1161.09 mg/L of β-elemene and 852.36 mg/L of germacrene A, which is the highest reported yield at shake flask level. In 4-L fed-batch fermentation, the production of β-elemene and germacrene A reached 3.52 g/L and 2.13 g/L, respectively.

Dependencies are inferred from the claim text.