New genetic engineering strategies that can increase the production of isoprene in Saccharomyces cerevisiae were explored. In order to increase the flux of Acetyl-CoA in the cytoplasm of S. cerevisiae, the genes MPC2, POR2, PDA1 were deleted separately and the vector pESC-His-ispS containing the Populus alba isoprene synthase gene was transformed into the above three knockout mutants respectively and obtaining engineered strains SC1, SC2, SC3. The result showed that SC3 had the best performance in shake-flasks, which was 0.3-fold higher than that of the original strain. To further increase the flux of Acetyl-CoA in the cytoplasm of S. cerevisiae, the 5-phosphate-specific phosphoketolase ( LmPK ) from Leuconostoc mesenteroides and phosphotransacetylase (CkPTA) from Clostridium kluyveri were introduced into △PDA1 strain, obtaining engineered strain SC4. And the vector pESC-His-ispS was transformed into SC4 to obtain pHB1. Subsequently, the vector pESC-His-ispS-EcIDI1 containing the E. coli IDI1 was transformed into SC4 to obtain engineered strain pHB2. The titer of isoprene reached 54 μg/L in pHB2 strain in shake flask fermentation, which was about 1-fold higher than that in the original strain. This study provides new idea for the subsequent research of isoprene production, and also lays foundation for the industrialization of isoprene production in the yeast.
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