UDP-glucosyl transferase enzymatic conversion of rebaudioside A to M
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Solution Overview
Problem
Current methods for producing rebaudioside M are inefficient, resulting in low purity and high costs due to the low content of rebaudioside M in Stevia rebaudiana plants and the presence of impurities in steviosides, limiting its commercialization and application.
Innovation Solution
A biological method using UDP-glucosyl transferase and recombinant cells to convert rebaudioside A or D into rebaudioside M through a glucosyl transfer reaction, employing a UDP-glucose regeneration system to reduce costs and achieve high-purity production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If rebaudioside M is extracted from Stevia rebaudiana plants, then the product is obtained naturally, but the content is very low and purification is required
Solution Approach 1:
The patent uses UDP-glucosyl transferase as a biological intermediary to catalyze the conversion of rebaudioside A or D into rebaudioside M. This enzyme-mediated transformation allows production without relying on low-yield plant extraction, thereby improving productivity while maintaining product quality through controlled enzymatic reaction conditions.
Solution Approach 2:
The patent optimizes reaction parameters including temperature (25-37°C), pH (6.0-7.5), and substrate concentration to maximize enzyme activity and rebaudioside M yield. By controlling these parameters, the enzymatic method achieves high conversion efficiency and production rates that cannot be obtained through natural extraction alone.
2Manufacturing precision
If rebaudioside A is used as substrate with UDP-glucose, then rebaudioside M is produced, but the cost is high
Solution Approach 1:
The patent implements a UDP-glucose regeneration system where UDP is recovered and reused through the action of sucrose synthase. This regeneration mechanism reduces the consumption of expensive UDP-glucose substrate, thereby lowering production costs while maintaining high product purity through controlled enzymatic reactions.
Solution Approach 2:
The enzymatic system is designed to be self-sustaining through the UDP-glucose regeneration cycle. The system recycles UDP back into UDP-glucose using sucrose and sucrose synthase, reducing the need for continuous addition of expensive substrates and enabling cost-effective large-scale production of high-purity rebaudioside M.
3Ease of manufacture
If traditional extraction method is used, then steviosides are obtained, but impurities are present affecting application
Solution Approach 1:
The patent selectively extracts only the desired rebaudioside M product through specific enzymatic transformation of rebaudioside A or D substrates. Unlike traditional extraction that co-extracts multiple steviosides and impurities, this method produces high-purity rebaudioside M with minimal impurities, expanding its application field in food and beverage industries.
4Ease of manufacture
If UDP-glucose regeneration system is employed, then production cost is reduced, but system complexity increases
Solution Approach 1:
The patent combines multiple enzymatic functions into an integrated system where UDP-glucosyl transferase, sucrose synthase, and other enzymes work together in a coordinated manner. This merged system achieves both cost reduction through substrate regeneration and manageable complexity by using naturally compatible enzymatic reactions that occur under the same physiological conditions.
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
This method significantly reduces production costs and time, enhances product purity, and expands the application field of rebaudioside M in food and beverage industries by utilizing microbial cells and enzyme catalysis.
Implementation Method 1
rebaudioside M is generated by means of reaction of the substrate under the catalysis of UDP-glucosyl transferase
Implementation Method 2
rebaudioside M is generated by means of reaction of the substrate under the catalysis of UDP-glucosyl transferase and/or recombinant cells containing the UDP-glucosyl transferase
Implementation Method 3
a UDP-glucose regeneration system composed of sucrose, sucrose synthetase and UDP
Data Source
AI summary
Provided is a method for preparing rebaudioside M by using an enzyme method. In the method, rebaudioside A or rebaudioside D is used as a substrate; and in the existence of a glucosyl donor, rebaudioside M is generated by means of reaction of the substrate under the catalysis of UDP-glucosyl transferase and/or recombinant cells containing the UDP-glucosyl transferase.