UGT-B Enzyme Converts Rebaudioside A to Sweeteners
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Solution Overview
Problem
The high cost and inefficiency of extracting and purifying rebaudioside D and M from Stevia leaves due to their low abundance and bitter taste limitations, which hinders their widespread use as natural sweeteners.
Innovation Solution
Development of uridine diphosphate (UDP)-glycosyltransferase B (UGT-B) enzyme that converts rebaudioside A into rebaudioside D and further into rebaudioside M, using a specific amino acid sequence, enabling their mass production through microbial expression and enzymatic reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If rebaudioside D and M are extracted and purified from Stevia leaves using traditional methods, then natural sweeteners with less bitter taste can be obtained, but the production cost is high and efficiency is low due to low abundance in leaves
Solution Approach 1:
The patent introduces UDP-glycosyltransferase enzymes as intermediary catalysts to convert rebaudioside A into rebaudioside D and M. This enzymatic conversion pathway serves as a mediator between the readily available rebaudioside A and the desired low-bitterness sweeteners, enabling efficient production without direct extraction from Stevia leaves.
Solution Approach 2:
The patent replaces traditional mechanical extraction and purification methods with enzymatic conversion. Instead of physically extracting rebaudioside D and M from plant material, the system uses biochemical reactions catalyzed by UDP-glycosyltransferase enzymes to synthesize these compounds from rebaudioside A, significantly improving production efficiency.
2Object-affected harmful factors
If rebaudioside D and M are extracted from Stevia leaves, then high-quality sweeteners with excellent taste can be produced, but the extraction and purification process requires high cost
Solution Approach 1:
UDP-glycosyltransferase enzymes act as biochemical intermediaries that catalyze the conversion of rebaudioside A to rebaudioside D and M. This enzymatic pathway provides a cost-effective route to produce high-quality sweeteners without the expensive extraction and purification processes required by traditional methods.
Solution Approach 2:
The patent changes the chemical parameters of rebaudioside A through enzymatic glycosylation reactions. By controlling the enzymatic conversion conditions, rebaudioside A is transformed into rebaudioside D and M with different chemical structures that exhibit reduced bitterness, providing a cost-effective method to improve product quality.
3Reliability
If traditional extraction methods are used to obtain rebaudioside D and M, then natural sweeteners can be produced, but the process is inefficient due to low abundance in Stevia leaves
Solution Approach 1:
The patent replaces inefficient mechanical extraction with efficient enzymatic synthesis. Instead of relying on the low natural abundance of rebaudioside D and M in Stevia leaves, the system uses UDP-glycosyltransferase enzymes to synthesize these compounds from abundant rebaudioside A, dramatically improving production efficiency while maintaining natural sweetener quality.
Solution Approach 2:
UDP-glycosyltransferase enzymes serve as reliable intermediary catalysts that ensure consistent production of natural sweeteners. This enzymatic conversion process provides a reliable and efficient pathway to produce rebaudioside D and M with controlled quality, independent of the variable natural abundance in Stevia leaves.
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
Facilitates the high-yield, high-purity production of rebaudioside D and M with minimal by-products, using inexpensive raw materials, thus overcoming the cost and efficiency challenges of traditional extraction methods.
Implementation Method 1
it has been newly found that a polypeptide sequence whose function was not previously revealed has a glycosyltransferase activity, and that the polypeptide has a glycosyltransferase activity of converting rebaudioside A into rebaudioside D
Implementation Method 2
reacting the rebaudioside D with nucleotide diphosphate to which glucose is bonded in the presence of uridine diphosphate (UDP)-glycosyltransferase A (UGT-A) to prepare rebaudioside M
Data Source
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AI summary
The present application relates to a novel uridine diphosphate (UDP)-glycosyltransferase B (UGT-B), a polynucleotide encoding the uridine diphosphate (UDP)-glycosyltransferase B (UGT-B), an expression vector containing the polynucleotide, a microorganism comprising the uridine diphosphate (UDP)-glycosyltransferase B (UGT-B) or a polynucleotide encoding the uridine diphosphate (UDP)-glycosyltransferase B (UGT-B), and a method for producing rebaudioside D and rebaudioside M using the microorganism.