YberC Transporter HMO Production
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Solution Overview
Problem
Current biotechnological methods for producing human milk oligosaccharides (HMOs) face challenges in efficiently identifying the right transporter proteins for effluxing different recombinantly produced HMO structures, due to unpredictable substrate specificity of sugar transporters, which hampers high-scale industrial manufacturing and downstream processing efficiency.
Innovation Solution
Genetically modified cells expressing a heterologous Major Facilitator Superfamily (MFS) transporter protein, specifically the YberC protein, are developed to enhance the production and efflux of HMOs, particularly tri- and tetra-unit oligosaccharides, thereby improving yields and reducing by-product formation and biomass during fermentation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If recombinant cells are engineered to express high levels of glycosyltransferases for HMO synthesis, then HMO production capacity increases, but metabolic burden on the cell increases and intracellular nucleotide sugar pool becomes depleted
Solution Approach 1:
The patent extracts the HMO synthesis pathway from the central carbon metabolism by utilizing the glyoxylate shunt pathway, which allows the cell to produce HMOs without depleting intracellular nucleotide sugar pools. This pathway separation enables sustained high-level HMO production while maintaining cellular metabolic balance.
Solution Approach 2:
The patent employs preliminary action by pre-establishing the glyoxylate shunt pathway and introducing specific enzymes (Isocitrate Lyase, Malate Synthase, PEP Carboxylase) before HMO synthesis begins. This preparatory metabolic infrastructure allows the cell to efficiently convert carbon sources into HMO precursors without depleting existing nucleotide sugar reserves.
2Adaptability or versatility
If multiple sugar transporter proteins are tested for effluxing different HMO structures, then substrate specificity coverage improves, but time and resources for identification and optimization increase
Solution Approach 1:
The patent identifies a universal approach by selecting transporter proteins with broad substrate specificity that can efflux multiple types of HMOs (trisaccharides, tetrasaccharides, and other oligosaccharides). This multi-functional transporter selection eliminates the need to test multiple specialized transporters for different HMO structures, significantly reducing optimization time while maintaining comprehensive coverage.
3Ease of manufacture
If conventional bacterial expression systems are used for HMO production, then cost efficiency and scalability are achieved, but inherent limitations in product yield and purity persist
Solution Approach 1:
The patent applies parameter changes by modifying key metabolic parameters in the bacterial expression system: introducing the glyoxylate shunt pathway, optimizing carbon source utilization, and adjusting enzyme expression levels. These parameter modifications enable conventional bacterial systems to achieve both cost efficiency/scalability and improved product yield/purity simultaneously.
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 use of YberC-expressing cells leads to increased production of specific HMOs like LNT-2, LNnT, and LNnT, with decreased by-product formation and biomass, facilitating more efficient recovery and purification, thus enhancing the overall manufacturing process.
Implementation Method 1
a method for recombinant production of human milk oligosaccharides (HMO) using a genetically modified cell expressing a protein of the major facilitator superfamily (MFS)... facilitating the efflux of synthesized HMOs into the extracellular media
Data Source
AI summary
The present inventive concept relates to a genetically modified cell enabled for the production of an oligosaccharide, preferably, an HMO, comprising a recombinant nucleic acid encoding a protein of the MFS superfamily; and methods using said cell for the production the oligosaccharide, preferably an HMO.