Construction of double promoter drived invertase Suc2 overexpression Saccharomyces cerevisiae strain
To improve the expression level of sucrose converting enzyme Suc2 in Saccharomyces cerevisiae,tandem double promoters were used to jointly initiate the expression of Suc 2 gene and an efficient strain for producing inulin based ethanol was constructed.A recombinant expression vector pYC230-DP-mSuc2 containing a strong promoter PGK1p composed of tandem phosphoglycerate kinase was constructed,which was transformed into Saccharomyces cerevisiae BY4741 by electric shock,to construct the recombinant strain BY-DP.Through real-time fluorescence quantitative PCR,high-performance gas chromatography and shake flask culture validation,it was demonstrated that the recombinant strain BY-DP had higher gene transcription levels of Suc2,enzyme activity and ethanol production after fermentation for 48 h in a medium containing 15％inulin,increased by 30.0％,18.6％,and 25.3％,respectively,compared with the control recombinant strain BY-mS containing a single PGK1p promoter.The expression of Suc2 was effectively enhanced,and the ability of recombinant bacteria to ferment inulin and produce ethanol was improved by constructing a tandem double PGK1p promoter.
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