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Figure 1.
Functional characterization of AmGT28 and AmGT44. (a) Glycosylation of 1 by recombinant AmGT28/AmGT44 to produce 1a using UDP-Glc as the sugar donor. (b) UHPLC/UV chromatograms of the enzymatic reaction mixture. Control, reactions conducted using boiled protein; STD, reference standard. (c) MS and MS/MS spectra of 1a in the positive ion mode from the reaction catalyzed by AmGT28.
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Figure 2.
Substrate specificity of AmGT28 and AmGT44. (a), (b) Conversion rates (%) of glycosylated products for substrates 1−15, using UDP-Glc as the sugar donor (n = 3). *, Products identified by comparing with reference standards; Δ, product purified from scaled-up reactions and characterized by NMR. (c) Structures of substrates 1−15.
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Figure 3.
Substrate preference of AmGT44. (a) Kinetic parameters of AmGT44 using maackiain (2) as the substrate and UDP-Glc as the sugar donor. (b) Kinetic parameters of AmGT44 using 7-hydroxyisoflavone (4) as the substrate and UDP-Glc as the sugar donor.
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Figure 4.
Whole cell catalytic reactions for AmGT28 and AmGT44. (a) Optimization of reaction conditions using methods 1−9. (b) Conversion rates of compound 2 using AmGT28 and AmGT44 under different conditions 1−9. Mn: Method number. (c), (d) Conversion rates of glycosylated products for substrates 1−15. For all experiments, n = 3.
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