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The rate-limiting step in the production of Cy7G is glucosylation catalyzed UGT88E18. To overcome this obstacle, the GSL enzyme SuSy from Glycine max was heterologously expressed and purified to homogeneity (Table S1). The reaction of SuSy with UGT88E18, which regenerates expensive UDPG by a reaction using cheap sucrose, yields Cy7G.
Calycosin and formononetin were transformed to Cy7G in the presence of 50, 150, and 200 mU mL-1 of UGT88E18 with varying amounts of SuSy. HPLC-ESI/MS was used to evaluate the resulting titers. The results showed that 95% of the calycosin could be converted to Cy7G by using the optimal ratios for UGT88E18, and SuSy.
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