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Publication Abstract from PubMed

The addition of the O-linked N-acetylglucosamine (O-GlcNAc) is a significant modification for active molecules, such as proteins, carbohydrates, and natural products. However, the synthesis of terpenoid glycoside derivatives decorated with GlcNAc remains a challenging task due to the absence of glycosyltransferases, key enzymes for catalyzing the transfer of GlcNAc to terpenoids. In this study, we demonstrated that the enzyme mutant UGT74AC1(T79Y/L48M/R28H/L109I/S15A/M76L/H47R) efficiently transferred GlcNAc from uridine diphosphate (UDP)-GlcNAc to a variety of terpenoids. This powerful enzyme was employed to synthesize GlcNAc-decorated derivatives of terpenoids, including mogrol, steviol, andrographolide, protopanaxadiol, glycyrrhetinic acid, ursolic acid, and betulinic acid for the first time. To unravel the mechanism of UDP-GlcNAc recognition, we determined the X-ray crystal structure of the inactivated mutant UGT74AC1(His18A/Asp111A) in complex with UDP-GlcNAc at a resolution of 1.66 A. Through molecular dynamic simulation and activity analysis, we revealed the molecular mechanism and catalytically important amino acids directly involved in the recognition of UDP-GlcNAc. Overall, this study not only provided a potent biocatalyst capable of glycodiversifying natural products but also elucidated the structural basis for UDP-GlcNAc recognition by glycosyltransferases.

Enzymatic Synthesis of Novel Terpenoid Glycoside Derivatives Decorated with N-Acetylglucosamine Catalyzed by UGT74AC1.,Li J, Li R, Shang N, Men Y, Cai Y, Zeng Y, Liu W, Yang J, Sun Y J Agric Food Chem. 2024 Jun 26;72(25):14255-14263. doi: 10.1021/acs.jafc.4c02832. , Epub 2024 Jun 12. PMID:38867497^[3]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.