The Biosynthesis Of Salicylic Acid In Saccharomyces Cerevisiae

Saccharomyces cerevisiae, a representative eukaryotic model, has been favored in recent years in biosynthesis and bio-fermentation.Salicylic acid is a intermediates widely used in the industry, how to achieve green production has great research value. In this study,synthetic salicylic acid pathway was transformed into Saccharomyces cerevisiae. With the synthetic biology, we used genetic engineering to improve the yield of salicylic acid in Saccharomyces cerevisiae.Specific work is as follows:Introducing exogenous gene entC and PfpchB in Saccharomyces cerevisiae BY4741, expressed successfully and catalyzed the synthesis of salicylic acid. We optimized the promoter, altered the expression of exogenous genes, and constructed plasmid 181-PGI1-EP,181-TEF2-EP, 181-TEF1-EP. The yield of salicylic acid of recombinant strain BY4741-181-TEF1-EP reached 12.06 ± 0.38 mg·L-1.Modification of shikimic acid pathway of Saccharomyces cerevisiae, remove the feedback inhibition of Aro4, and we optimize the promoter construct plasmid 181 -PGI1 -EPA ,181-TEF2-EPA ,181 -TEF1-EPA. The yield of S. cerevisiae BY4741-181-TEF1-EPA was 33.2±0.3 mg L-1. We integrated and replaced the promoter Arol for TEF1p.The yield of salmonic acid of the recombinant strain BY4741-TEF1p-Arol-181-TEF1-EPA was 7.76 ± 0.41 mg·L-1.Expression of Escherichia coli Shikimic Pathway in Saccharomyces cerevisiae. The plasmid is divided into three modules:module one is 181-TEF1-EPAC, 195-AroC, module two is 195-AL,module three is 195-BDE, module two and module three co-expressed 195-ALBDE. The yield of salicylic acid was 41.68 ± 1.92 mg·L-1.Overexpressing genes in the pentose phosphate pathway. The single gene overexpression plasmids were 195-ZWF1, 195-SOL3,195-GND1, 195-TKL1, 195-TAL1, and the polygene overexpression plasmid was 195-ZWF1-SOL3, 195-SOL3-GND1, 195-ZWF1-GND1,195-TKL1-TAL1 ,195-ZWF1-SOL3-GND1. The highest yield of salicylic acd d was 46.71 ± 0.27 mg·L-1 when overexpressing GND1 In the overexpression of polygenes, the co-expression of ZWF1 and GND1 was the highest, which was 46.15 ± 0.80 mg·L-1.