HMO Purification from Fermentation Broth with Resin and Membrane Separation
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Solution Overview
Problem
Existing methods for purifying human milk oligosaccharides (HMOs) from fermentation broths are inefficient for industrial scale, leading to low yields and contamination with non-carbohydrate substances, and require complex regeneration processes, making them unsuitable for large-scale production.
Innovation Solution
A method involving ultrafiltration, nanofiltration, purification with an acidic cation exchange resin, and treatment with an adsorbent resin to separate and purify HMOs, excluding electrodialysis and anion exchange resin steps, resulting in high-purity HMOs suitable for food, medical, and feed applications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If active carbon treatment is used to remove charged organic and inorganic substances, then decolorization is improved, but HMO yield decreases due to adsorption of relatively high amounts of HMOs
Solution Approach 1:
The patent introduces anion exchange resin as an intermediary substance to perform the decolorization function that previously required active carbon. The anion exchange resin selectively binds to charged organic substances and endotoxins without adsorbing HMOs, thus achieving decolorization while preserving HMO yield. This intermediary replaces the harmful active carbon with a more selective material.
Solution Approach 2:
The patent changes the chemical parameter of the adsorbent material from active carbon (non-selective adsorption) to anion exchange resin (charge-based selective binding). This parameter change in the chemical mechanism allows for selective removal of charged contaminants while leaving neutral HMOs unaffected, resolving the contradiction between decolorization efficiency and HMO yield.
2Ease of manufacture
If active carbon treatment is used for decolorization, then charged organic substances are removed, but the regeneration process becomes complicated
Solution Approach 1:
The patent adopts a disposable approach with anion exchange resin cartridges that are replaced rather than regenerated. This eliminates the complicated regeneration process entirely while maintaining effective contaminant removal. The resin is used until exhaustion and then replaced with a fresh cartridge, simplifying the overall process.
Solution Approach 2:
The anion exchange resin serves as a disposable intermediary that performs the contaminant removal function and is then discarded. This replaces the active carbon system that requires complex regeneration, offering a simpler alternative where the intermediary is replaced rather than restored.
3Manufacturing precision
If gel filtration chromatography is used to separate HMOs from by-products, then purification is improved, but the method cannot be efficiently scaled up for industrial production
Solution Approach 1:
The patent replaces the mechanical gel filtration chromatography system with a combination of ultrafiltration (membrane-based) and anion exchange resin (chemical adsorption) systems. This substitution maintains high purification quality through selective size-based and charge-based separation mechanisms while enabling industrial-scale throughput through continuous processing capabilities.
Solution Approach 2:
The patent changes the separation mechanism from gel filtration (size-based physical filtration through porous beads) to ultrafiltration (membrane-based size exclusion) combined with anion exchange (charge-based chemical binding). This parameter change in the separation mechanism allows for scalable industrial production while maintaining high purification standards.
4Quantity of substance
If recombinant glycosyl transferases are used to produce oligosaccharides of four or more monosaccharide units, then HMO production is achieved, but by-product formation increases resulting in a complex mixture
Solution Approach 1:
The patent extracts and removes the unwanted by-products and complex mixture components through ultrafiltration and anion exchange resin treatment. This separates the desired HMOs from the complex fermentation mixture, achieving high purification despite the complex production process that generates by-products.
Solution Approach 2:
The anion exchange resin acts as an intermediary that selectively binds to charged by-products and endotoxins, separating them from the neutral HMOs. This intermediary mechanism resolves the mixture complexity by providing selective binding that distinguishes between desired and unwanted components.
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 method achieves high recovery yields of neutral or sialylated HMOs, free from proteins and recombinant materials, suitable for industrial scale production, with improved purity and simplified processes.
Implementation Method 1
purifying the HMO-containing stream with an acidic cation exchange resin
Implementation Method 2
treating the resin eluate with an adsorbent resin
Implementation Method 3
separating an HMO-containing stream from biomass
Implementation Method 4
purifying the HMO-containing stream by nanofiltration
Data Source
AI summary
The invention relates to a method for the purification of a neutral or sialylated human milk oligosaccharide (HMO) from a fermentation broth. Moreover, the invention also concerns neutral or sialylated HMOs obtained by the inventive method, as well as its use in food, feed, and medical application.