Constitutive expression and characteristics of glycosyltransferase UGT76G1 in Saccharomyces cerevisiae
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Graphical Abstract
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Abstract
In this study, a new expression vector p ESCD which contains the two constitutive promoters, p PGK1 and p TEF1 that replaced the original induced promoter GAL1 / GAL10 in p ESC- Leu, was designed and constructed.The synthetic glycosyltransferase UGT76G1 coding gene was inserted into the restriction sites, Sal I and Xho I of p ESCD to construct the recombinant plasmid p ESCD- UGT, which was then transformed into the Saccharomyces cerevisiae strain YPH499.The results confirmed that the recombinant cells grew into the stable phase when cultured for 24 h and the highest expression of the recombinant protein was observed when cultured for 8h in SD- L.In case of 1% methylbenzence as the cell permeating agent, 288 mg / L RA was produced after reaction for 15 h using the whole- cell catalyst, which was 5 times higher than that of thecontrol group.
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