Rebaudioside D and M Production via Enzymatic Glucosylation
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Solution Overview
Problem
Current methods for producing rebaudioside D and rebaudioside M are inefficient and costly, failing to achieve the desired taste and flavor profiles similar to sucrose, and require high-purity starting materials, making them unsuitable for commercial use.
Innovation Solution
A method involving the use of recombinant microorganisms expressing EUGT11 and UGT76G1 enzymes to convert rebaudioside A and stevioside into rebaudioside D and M, with the inclusion of sucrose and trisodium citrate, followed by purification through heating, ultra-filtration, nanofiltration, and crystallization, to produce a sweetener composition with a desired taste profile.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If conventional methods are used to produce rebaudioside D and M, then production costs are high and efficiency is low, but the taste profile does not resemble sucrose and high-purity starting materials are required
Solution Approach 1:
The patent applies parameter changes by modifying the chemical structure of rebaudioside D and M through controlled glucosylation reactions. By adjusting reaction conditions (temperature, pH, enzyme concentration) and substrate ratios, the invention optimizes both production efficiency and taste profile, achieving a balance between manufacturing ease and product quality without requiring high-purity starting materials.
Solution Approach 2:
The patent uses an intermediary approach by introducing specific enzymes (glucosyltransferases) as mediators to convert rebaudioside A and stevioside into rebaudioside D and M. These enzymes act as catalysts that enable the transformation under mild conditions, reducing production costs while maintaining control over the molecular structure to achieve desired taste characteristics.
2Manufacturing precision
If high-purity starting materials are used, then product purity is improved, but production cost increases and method complexity increases
Solution Approach 1:
The patent applies preliminary action by performing selective glucosylation reactions that directly form the desired rebaudioside D and M structures from readily available starting materials (rebaudioside A and stevioside). This preliminary chemical modification eliminates the need for subsequent complex purification steps and high-purity starting materials, reducing both cost and complexity while maintaining product purity.
Solution Approach 2:
The patent extracts the essential function of high-purity starting materials by using readily available rebaudioside A and stevioside as substrates. Through selective enzymatic glucosylation, the invention extracts only the necessary structural modifications to achieve pure rebaudioside D and M, eliminating the need for expensive high-purity precursors while maintaining product quality.
3Ease of manufacture
If conventional production methods are used, then existing processes are simple, but they fail to achieve desired taste characteristics and flavor profiles
Solution Approach 1:
The patent applies local quality by making targeted modifications to specific positions in the steviol glycoside molecular structure through selective glucosylation. By controlling where glucose units are added (at specific hydroxyl positions), the invention locally alters the molecular properties to achieve desired taste and flavor characteristics while maintaining overall process simplicity.
Solution Approach 2:
The patent replaces complex mechanical separation and purification systems with a biochemical approach using enzymatic glucosylation. This substitution allows for precise control of molecular structure through chemical reaction conditions rather than physical separation, achieving superior taste profiles while maintaining process simplicity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method enables the production of rebaudioside D and M with a taste similar to sucrose, achieving high purity and low production costs, addressing the inefficiencies and cost issues of existing methods while maintaining natural characteristics without introducing synthetic components or energy.
Implementation Method 1
incubating the starting composition with a recombinant microorganism in a mixture, wherein the recombinant microorganism expresses an EUGT11 enzyme and a UGT76G1 enzyme
Implementation Method 2
purifying rebaudioside D and/or rebaudioside M from the mixture
Implementation Method 3
purification through heating, ultra-filtration, nanofiltration, and crystallization
Implementation Method 4
purification through heating, ultra-filtration, nanofiltration, and crystallization
Implementation Method 5
purification through heating, ultra-filtration, nanofiltration, and crystallization
Data Source
AI summary
The invention relates to methods for producing rebaudioside D and/or rebaudioside M, and compositions comprising the same. The invention provides a method for producing RD and/or RM compositions. The method comprises using rebaudioside A and/or stevioside as substrate and a recombinant microorganism or an enzyme produced by the recombinant microorganism or a metabolite of the recombinant microorganism to catalyze the reaction of the substrate in the presence of sucrose and trisodium citrate and produce a mixture of rebaudioside D and rebaudioside M, and then separates and purifies the mixture to obtain rebaudioside D or rebaudioside M.