Abstract
The macrolactone rapamycin (RAP) presents a broad range of bioactivities, but its clinical applications are compromised due to the poor water solubility and low bioavailability, which could probably be overcome by glycosylation. In this study, we tested a set of promiscuous glycosyltransferases (GTs) to modify rapamycin with four different sugar donors. BsGT-1 displayed the best glycosylation activity with a preference for UDP-glucose, and the glycosylation happened at C-28 or C-40 of rapamycin, producing rapamycin-40-O-β-d-glucoside (RG1), and two new compounds rapamycin-28-O-β-d-glucoside (RG2) and rapamycin-28,40-O-β-d-diglucoside (RG3). The glycosylation remarkably improved water solubility and almost completely abolished cytotoxicity but simultaneously attenuated the antifungal, antitumor, and immunosuppression bioactivities of rapamycin. We found the glycosylation at C-40 had less effect on the bioactivities than that at C-28. The molecular docking analysis revealed that the glycosylation, especially the glycosylation at C-28, weakened the hydrophobic and hydrogen bonding contacts between the rapamycin glucosides and the binding proteins: the FK506-binding protein (FKBP12) and the FKBP12-rapamycin binding (FRB) domain. This study highlights a succinct approach to expand the chemical diversity of the therapeutically important molecule rapamycin by using promiscuous glycosyltransferases. Moreover, the fact that glycosyl moieties at different positions of rapamycin affect bioactivity to different extents inspires further glycosylation engineering to improve properties of rapamycin.
Key points
• Rapamycin was glycosylated efficiently by some promiscuous GTs.
• Glycosylation improved water solubility, attenuated cytotoxicity, and bioactivities.
• Glycosylation affected the interactions between ligand and binding proteins.
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Funding
This work was financially supported by the National Key Research and Development Programs of China (nos. 2018YFA0900400 and 2018YFA0901704) and the National Natural Science Foundation of China (NSFC) (nos. 31670076 and 31471183) to Y.-Z.L, and NSFC (nos. 31900042 and 81973215) and the National Key Research and Development Program of China (2019YFA0905700) to C.S.W.
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P.Z., C.S.W., and Y.Z.L. designed researches; P.Z. and L.J.Z. performed researches; P.Z., X.J.Y., C.S.W., and Y.Z.L. analyzed data; Y.J.T. contributed analytical tools; P.Z., C.S.W., and Y.Z.L. wrote the paper; Y.Z.L. and C.S.W. provided funds for the project. All authors read and approved the manuscript.
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Zhang, P., Zhang, L., Yue, Xj. et al. Effects of glycosylation on the bioactivity of rapamycin. Appl Microbiol Biotechnol 104, 9125–9134 (2020). https://doi.org/10.1007/s00253-020-10895-3
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DOI: https://doi.org/10.1007/s00253-020-10895-3