Enzymatic production of sweet-tasting trisaccharides as natural dietary fiber

Overview

This solution leverages levansucrase enzymes to produce sweet-tasting trisaccharides such as lactosucrose, cellobiofructose, and erlose from natural and sustainable resources. Unlike conventional sugars, these trisaccharides are expected to qualify as dietary fiber due to their chain length and specific linkage type, while delivering a sweet taste profile. The technology offers food, beverage, and nutraceutical manufacturers a pathway to reformulate products with reduced sugar content while adding prebiotic health benefits. By using biocatalytic fructosylation rather than chemical synthesis, the process aligns with consumer demand for clean-label, naturally derived ingredients.

Technical specifications

Key features:

• Recombinant levansucrase produced heterologously in Escherichia coli with very high yields
• Volumetric enzyme activity of 300,000 Units per liter of culture
• Purified enzyme specific activity of 300 U/mg
• Exclusive fructosyl transfer to alternative acceptor molecules (lactose, maltose, cellobiose, erlose) with no detectable levan-type fructan byproducts
• HPLC-validated product purity, confirming absence of polymeric levan or levan-type FOS
• Process adaptable to fed-batch and continuous flow cultivation strategies
• Potential for immobilized enzyme formats to enhance reusability and process economics
• Compatible purification workflows including ultrafiltration, nanofiltration, and microbial fermentation

Technology readiness level

The core biocatalyst has been validated through recombinant production, purification, and detailed enzyme characterization using biological triplicates. Initial fructosylation reactions with lactose, erlose, and maltose have been confirmed at laboratory scale, with HPLC analysis verifying clean product profiles. Next-stage validation will focus on scaling enzyme production, optimizing cultivation strategies, establishing robust purification processes, and conducting studies on the prebiotic effects of purified trisaccharides on probiotic microorganisms and human gut microbiota members. The technology is currently at a laboratory-validated stage (TRL 4) progressing toward pilot-scale demonstration.