We established enzymatic method for specific determination of stevioside content. The key enzyme came from recombinant β-glucosidase BT_3567 (rBT_3567) from Bacteroides thetaiotaomicron HB-13 which exhibited selective hydrolysis of stevioside at β-1,2-glycosidic bond to yield rubusoside and glucose. Released glucose was detected with glucose oxidase and peroxidase allowed indirect quantitation of stevioside content in samples by using a colorimetric-based approach. The series of reactions for stevioside determination can be completed within 1 h at 37 °C. Stevioside determination using the enzymatic assay strongly correlated with results obtained from HPLC with high specificity and accuracy. This analysis demonstrates that the enzymatic method developed in this study is specific, easy to perform, accurate, and yields reproducible results.
Reference
Udompaisarn S, Arthan D, Somana J. Development and validation of an enzymatic method to determine stevioside content from Stevia rebaudiana. Journal of Agricultural Food Chemistry 2017, 65(15) 3223-3229.
| Relevant SDGs | |
|---|---|
 |
|
| BC investigator | |
 Asst. Prof. Jamorn Somana |
|