Recombinant Microorganisms for Scalable Steviol Glycoside Fermentation
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Solution Overview
Problem
Existing methods for producing steviol glycosides from Stevia plants are inefficient, vary with agricultural conditions, and require substantial resources, while commercial demand for high-potency, natural sweeteners with improved taste profiles is increasing.
Innovation Solution
A recombinant microorganism with a deficiency in serine/threonine protein kinase and expressing uridine diphosphate-dependent glucosyltransferase (UGT) activity is used to enhance the production of steviol glycosides like rebaudioside M and rebaudioside D through fermentation or bioconversion processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If steviol glycosides are extracted from Stevia plants, then natural sweeteners are obtained, but extraction yields vary with agricultural and environmental conditions and require substantial land area and resources
Solution Approach 1:
The patent replaces mechanical extraction from plants with a biological production system using recombinant microorganisms. The microorganisms express UGT enzymes that catalyze the glucosylation of steviol to produce steviol glycosides, substituting agricultural extraction with controlled fermentation processes that eliminate variability from environmental and agricultural conditions
Solution Approach 2:
The patent modifies the microorganism's physiological parameters by creating a deficiency in serine/threonine protein kinase activity. This parameter change redirects metabolic flux toward the production of target steviol glycosides, improving both yield consistency and production efficiency compared to plant extraction
2Productivity
If conventional fermentation methods are used to produce steviol glycosides, then production scalability is achieved, but the taste profile contains bitter after-taste and production of specific glycosides like rebaudioside M is limited
Solution Approach 1:
The patent applies local quality by creating a specific deficiency in serine/threonine protein kinase activity within the microorganism. This localized metabolic modification preferentially directs flux toward rebaudioside M production while maintaining overall fermentation scalability, achieving both high productivity and precise glycoside composition control
Solution Approach 2:
By changing the kinetic parameters of the metabolic pathway through protein kinase deficiency, the system achieves preferential production of rebaudioside M with improved taste profile, while maintaining the scalability benefits of fermentation processes
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 recombinant microorganism improves the yield and quality of steviol glycosides, providing a standardized, clean source of high-potency sweeteners with reduced after-taste, meeting growing commercial demand for natural sweeteners.
Implementation Method 1
comprises a polynucleotide encoding a polypeptide having uridine diphosphate-dependent glucosyltransferase (UGT) activity
Implementation Method 2
produce at least the steviol glycosides such as rebaudioside A, rebaudioside D and rebaudioside M by fermentation
Data Source
AI summary
The invention disclosed herein relates generally to the field of recombinant production of a steviol glycoside, to the field of bioconversion of steviol into a steviol glycoside and to the field of bioconversion of a steviol glycoside into a further steviol glycoside. Particularly, the invention provides a process for recombinant production of a steviol glycoside, a process of bioconversion of steviol into a steviol glycoside, a process for bioconversion of a steviol glycoside into a further steviol glycoside and a composition comprising a steviol glycoside. More particularly, the invention relates to a microorganism that has a deficiency of a serine/threonine protein kinase and comprises a polynucleotide encoding a polypeptide having uridine diphosphate-dependent glucosyltransferase (UGT) activity.