Review on Stevia Rebaudiana: A Comprehensive Analysis of its Properties and Applications

The perennial plant Stevia rebaudiana, also referred to as stevia, is indigenous to South America, especially Brazil and Paraguay. It is a member of the Asteraceae family. Stevia, a natural, calorie-free sweetener, has drawn a lot of attention in recent decades and is frequently promoted as a healthy substitute for sugar and artificial sweeteners. The purpose of this review article is to present a thorough examination of the characteristics, health advantages, extraction processes, and uses of stevia in a variety of industries, with a focus on food and drink. Steviol glycosides (SGs) are low in calories and high in sweetness, thus creating SGs with better flavor profiles is a major priority. Because of its increased sweetness, rebaudioside M8 (Reb M8), a novel non-natural SG derivative made by glycosylating rebaudioside D (Reb D) at the C-13 position with glycosyltransferase UGT94E13, shows promise for future development. However, additional research and commercialization are hampered by UGT94E13's low catalytic activity. By using a semi-rational design, this work sought to increase the enzymatic activity of UGT94E13. A variation, UGT94E13-F169G/I185G, was produced with a 13.90-fold increase in catalytic activity. Uridine diphosphate glucose was recycled using a cascade reaction including UGT94E13-F169G/I185G and sucrose synthase, which produced an effective preparation of Reb M8 with a 98% yield. Additionally, in accordance with the using molecular dynamics simulations to analyze the distances between the substrate Reb D and the enzymes and between Reb D and the glucose donor, it is discovered that the enhanced catalytic activity of UGT94E13-F169G/I185G is due to the beneficial effect of reducing the distance on glycosylation reaction activity. As a result, this study tackles the obstacle to the effective manufacturing of Reb M8 and lays the groundwork for its extensive use in the food sector.