Class
Article
College
College of Engineering
Department
Biological Engineering Department
Faculty Mentor
Jixun Zhan
Presentation Type
Poster Presentation
Abstract
Quercetin is a naturally occurring phenolic compound that is commonly used due to its various biological activities, but its low aqueous solubility and poor bioavailability hinder further clinical applications. For assessing the effect of the glycosylation of quercetin on its aqueous solubility, one glucosyltransferase gene (BbGT) was cloned from the genome of the strain Beauveria bassiana ATCC 7159 and heterologously expressed in different microorganism host cells, namely Escherichia coli, Saccharomyces cerevisiae, Pseudomonas putida, and Pichia pastoris. Then, the glycosyltransferase was purified, and its glycosylation capacity toward quercetin was verified. The result showed that the purified BbGT (50.4 kDa) could convert quercetin into different glucosidic derivatives. Then, the main structures of the derivatives were characterized as quercetin-7-O-β-D-glucoside and quercetin-3-O-β-D-glucoside by nuclear magnetic resonance (NMR) spectroscopy. In vitro enzyme assays showed that the optimal catalyzing temperature and PH conditions are 35℃ and 8.0 respectively. Meanwhile, the glucosidic activity of BbGT was stimulated by Ca2+, Mg2+, and Mn2+ but inhibited by Zn2+. Furthermore, we found that BbGT enzyme has flexible sugar-acceptor substrate specificity in the substrate specificity experiment, which include curcumin, resveratrol, and zearalenone. Interestingly, different numbers of glucose moieties can be added to the sugar-acceptor substrate curcumin and the glucose can be attached at different chemical group positions of resveratrol and zearalenone. Finally, the BbGT-harboring E. coli strain and S. cerevisiae were used as whole-cell biocatalysts to produce glucosidic derivatives from quercetin. Our findings effectively apply the enzymatic approach to obtain glucoside derivatives with enhanced solubility, which provides a helpful molecular method for synthetic biology and contributes to drug discovery. Presentation Time: Wednesday, 2-3 p.m.
Location
Logan, UT
Start Date
4-10-2021 12:00 AM
Included in
Identification, Isolation and Characterization of a Novel Glucosyltransferase from Beauveria bassiana
Logan, UT
Quercetin is a naturally occurring phenolic compound that is commonly used due to its various biological activities, but its low aqueous solubility and poor bioavailability hinder further clinical applications. For assessing the effect of the glycosylation of quercetin on its aqueous solubility, one glucosyltransferase gene (BbGT) was cloned from the genome of the strain Beauveria bassiana ATCC 7159 and heterologously expressed in different microorganism host cells, namely Escherichia coli, Saccharomyces cerevisiae, Pseudomonas putida, and Pichia pastoris. Then, the glycosyltransferase was purified, and its glycosylation capacity toward quercetin was verified. The result showed that the purified BbGT (50.4 kDa) could convert quercetin into different glucosidic derivatives. Then, the main structures of the derivatives were characterized as quercetin-7-O-β-D-glucoside and quercetin-3-O-β-D-glucoside by nuclear magnetic resonance (NMR) spectroscopy. In vitro enzyme assays showed that the optimal catalyzing temperature and PH conditions are 35℃ and 8.0 respectively. Meanwhile, the glucosidic activity of BbGT was stimulated by Ca2+, Mg2+, and Mn2+ but inhibited by Zn2+. Furthermore, we found that BbGT enzyme has flexible sugar-acceptor substrate specificity in the substrate specificity experiment, which include curcumin, resveratrol, and zearalenone. Interestingly, different numbers of glucose moieties can be added to the sugar-acceptor substrate curcumin and the glucose can be attached at different chemical group positions of resveratrol and zearalenone. Finally, the BbGT-harboring E. coli strain and S. cerevisiae were used as whole-cell biocatalysts to produce glucosidic derivatives from quercetin. Our findings effectively apply the enzymatic approach to obtain glucoside derivatives with enhanced solubility, which provides a helpful molecular method for synthetic biology and contributes to drug discovery. Presentation Time: Wednesday, 2-3 p.m.