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Enzymatic Synthesis of Novel Terpenoid Glycoside Derivatives Decorated with N -Acetylglucosamine Catalyzed by UGT74AC1.
- Source :
-
Journal of agricultural and food chemistry [J Agric Food Chem] 2024 Jun 26; Vol. 72 (25), pp. 14255-14263. Date of Electronic Publication: 2024 Jun 12. - Publication Year :
- 2024
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Abstract
- 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 <superscript>T79Y/L48M/R28H/L109I/S15A/M76L/H47R</superscript> 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 <superscript>His18A/Asp111A</superscript> in complex with UDP-GlcNAc at a resolution of 1.66 Å. 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.
- Subjects :
- Plant Proteins chemistry
Plant Proteins metabolism
Plant Proteins genetics
Biocatalysis
Acetylglucosamine chemistry
Acetylglucosamine metabolism
Glycosides chemistry
Glycosides metabolism
Glycosyltransferases metabolism
Glycosyltransferases chemistry
Glycosyltransferases genetics
Terpenes chemistry
Terpenes metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1520-5118
- Volume :
- 72
- Issue :
- 25
- Database :
- MEDLINE
- Journal :
- Journal of agricultural and food chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 38867497
- Full Text :
- https://doi.org/10.1021/acs.jafc.4c02832