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Computer-Guided Engineered Endo- and Exocleaving Glycosidase for Significantly Improving Production of Ginsenoside F1
Shi-Kun Lin1, Jinlin Zhou2, Yujing Lu3
1College of Food Science, South China Agricultural University, Guangzhou 510640, China.
Journal of Agricultural and Food Chemistry
|November 13, 2024
Summary
Researchers engineered a beta-glucosidase enzyme (BglSK) to specifically produce rare Ginsenoside F1 from Panax notoginseng. Protein engineering significantly enhanced enzyme activity, offering a new tool for producing valuable compounds.
Area of Science:
- Biochemistry
- Enzymology
- Natural Products Chemistry
Background:
- Ginsenosides in Panax notoginseng are extensively glycosylated, necessitating precise deglycosylation for isolating valuable compounds like Ginsenoside F1.
- Ginsenoside F1 is a rare and valuable compound with significant health benefits, making its efficient production a key objective.
Purpose of the Study:
- To identify and engineer a beta-glucosidase enzyme capable of specifically producing Ginsenoside F1.
- To enhance the catalytic efficiency of the identified enzyme through protein engineering for improved production yields.
Main Methods:
- Identification of beta-glucosidase BglSK with dual endo- and exoglycosidase activities on multi-6-O-glycosides.
- Protein engineering of BglSK to create the variant BglSK T137A/L508A.
- Characterization of enzyme kinetics (kcat/KM) for wild-type and engineered enzymes.
Main Results:
- The engineered BglSK T137A/L508A variant showed significantly enhanced catalytic efficiency.
- kcat/KM values for ginsenoside Rg1 and notoginsenoside R1 increased by 13.88-fold and 108.56-fold, respectively, compared to BglSK WT.
- Reduced steric hindrance and increased loop stability in the mutant likely contribute to enhanced activity.
Conclusions:
- The engineered BglSK T137A/L508A provides a powerful tool for the specific production of Ginsenoside F1.
- This study deepens the understanding of glycosidase activity enhancement mechanisms.
- The findings offer a pathway for producing high-value Panax notoginseng products.
