1837-LB: Designer Protein Sweelin, a Novel Hypersweet, Extremophile Protein as a Sugar Substitute

Introduction and Objective: sweelin^®, a novel sweet protein, ~3,000 sweeter than sugar enables up to 70% sugar reduction in numerous applications without compromising taste as assessed by professional sensory panels. The protein was designed using Agile-Integrative Computational Protein Design (AI-CPD) to improve stability and sensory profile of the monellin protein found in African fruits. It is produced via precision fermentation resulting with highly purified protein. Unlike small-molecule sweeteners, the protein is not expected to exhibit adverse effects, yet its digestibility required elucidation which was conducted via in vitro digestion models recreating gastric and intestinal proteolysis.

Methods: Digestibility of sweelin^® was studied using a computer controlled in vitro semi-dynamic digestion model that mimics the gastrointestinal conditions of a healthy adult via 2 consecutive steps, a dynamic gastric phase and a small intestinal (duodenal) phase that rely on an adjusted version of the INFOGEST in vitro digestion protocol. Digesta samples were collected at different timepoints of gastric and intestinal digestion and subsequently analyzed by SDS PAGE, with Coomassie Blue stain or Western blotting. Bioaccessible peptides were analyzed by LC-MS/MS.

Results: sweelin^® was readily digestible under intestinal conditions where bile and pancreatic secretions were added. In fact, the presence of physiological levels of trypsin and chymotrypsin led to the degradation of >95% of sweelin®. LC-MS/MS analysis of bioaccessible peptides reaffirmed that sweelin® is readily digested in the intestine into very short peptides. After 2h of intestinal digestion undigested protein levels were estimated to be <3% of the total ingested dose.

Conclusions: sweelin^® is a novel hyper-sweet designer protein shown to be effectively digested in the small intestine, much like commonly consumed proteins. Clinical trials are in progress to demonstrate no effect on diabetes-related biomarkers.