Sequential Fermentation for Oligosaccharide Purity
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Solution Overview
Problem
Current methods for producing prebiotic oligosaccharides, particularly human milk oligosaccharides, face challenges such as high costs, low purity, and inefficiencies due to the formation of unwanted by-products during fermentative production, making large-scale industrial production economically unfeasible.
Innovation Solution
A process involving genetically engineered microbial cells that utilize intermediate oligosaccharides with at least three monosaccharide moieties, allowing for sequential fermentation steps to split biosynthetic processes across different cells and compartments, reducing by-product formation and enhancing purity through specific biosynthetic and degradation reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional fermentative production methods are used to produce oligosaccharides, then production cost and scalability are improved, but purity is worsened due to formation of unwanted by-products
Solution Approach 1:
The patent divides the oligosaccharide production process into multiple sequential fermentation steps, with each step performed by a different genetically engineered microbial cell. The first step produces intermediate oligosaccharides (3-6 monosaccharide units), which are then used as substrates in subsequent steps to produce the final desired oligosaccharide. This segmentation prevents by-product formation at early stages and ensures high purity in the final product.
Solution Approach 2:
The patent employs preliminary fermentation steps to produce intermediate oligosaccharides before the final desired oligosaccharide is synthesized. These intermediate products serve as purified substrates for the subsequent fermentation step, ensuring that only the desired pathway proceeds while unwanted by-products are eliminated in earlier stages.
2Manufacturing precision
If multi-step synthesis approaches are used to produce specific oligosaccharides, then purity is improved, but production time and complexity are worsened
Solution Approach 1:
The patent merges multiple biosynthetic steps into a single continuous fermentation process. Different genetically engineered microbial cells are cultivated sequentially in the same fermentation system, with the output of one step serving as the input for the next step. This integration eliminates the need for separate purification steps between stages, maintaining high purity while reducing overall production time.
Solution Approach 2:
The patent implements continuous fermentation where intermediate oligosaccharides produced in one step are immediately used as substrates in the next step without interruption or purification. This continuous process maintains high productivity while ensuring purity through the sequential action of specialized microbial cells.
3Device complexity
If conventional single-step fermentation is used, then production simplicity is maintained, but by-product formation increases and purity decreases
Solution Approach 1:
The patent segments the fermentation process into multiple steps, with each step performed by a different genetically engineered microbial cell specialized in producing specific intermediate or final oligosaccharides. This segmentation allows each step to be optimized for its specific function, preventing by-product formation and ensuring high purity while maintaining reasonable process complexity.
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 approach enables the production of desired oligosaccharides with higher purity and reduced by-products, making the process more cost-effective and scalable for industrial applications.
Implementation Method 1
sequential fermentation steps to split biosynthetic processes across different cells and compartments
Implementation Method 2
an enzyme which is able to transfer a monosaccharide moiety form a donor substrate to the oligosaccharide consisting of at least three monosaccharide moieties
Data Source
AI summary
Disclosed is a process for the production of a desired oligosaccharide, the process comprises providing a genetically engineered microbial cell which possesses a saccharide importer for the uptake of an intermediate oligosaccharide, and an enzyme being able to convert the intermediated oligosaccharide by transferring a monosaccharide moiety from a donor substrate to the intermediate oligosaccharide; cultivating the genetically engineered microbial cell in the presence of an intermediate oligosaccharide to generate the desired oligosaccharide; and retrieving the desired oligosaccharide.