Oligosaccharide Purification from Fermentation Broth Without Electrodialysis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for isolating and purifying N-acetylglucosamine containing neutral oligosaccharides, such as lacto-N-triose II (LNT) and lacto-N-neotetraose (LNnT), from fermentation broths are complex and expensive, making them unsuitable for industrial-scale production.
Innovation Solution
A method involving ultrafiltration, nanofiltration, and treatment with ion exchange resins, specifically strongly acidic cation and weakly basic anion exchange resins, is used to separate N-acetylglucosamine containing oligosaccharides from non-carbohydrate compounds in a fermentation broth, without electrodialysis, resulting in a highly enriched solution with high yield and purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If gel filtration chromatography is used to separate oligosaccharides from fermentation broth, then purification quality is improved, but device complexity and cost increase significantly
Solution Approach 1:
The purification process is divided into multiple sequential steps: ultrafiltration to remove biomass, nanofiltration to concentrate oligosaccharides and remove salts, and ion exchange chromatography for final purification. Each step targets specific impurities, breaking down the complex purification task into manageable segments that can be performed with simpler, more scalable equipment.
Solution Approach 2:
The process exploits differences in molecular weight, charge, and solubility parameters to separate oligosaccharides from fermentation broth components. Ultrafiltration uses molecular weight cutoff, nanofiltration uses size exclusion, and ion exchange uses charge properties. By changing these parameters at different stages, effective separation is achieved without requiring complex chromatographic systems.
2Adaptability or versatility
If multiple types of glycosyl transferases are used to produce oligosaccharides of four or more monosaccharide units, then product diversity is improved, but separation and purification difficulty increases
Solution Approach 1:
Ion exchange resins act as intermediaries that selectively bind charged impurities and excess enzymes while allowing neutral oligosaccharides to pass through. This mediator approach simplifies the separation of complex mixtures produced by multiple glycosyl transferases, enabling purification without requiring complex chromatographic systems.
3Manufacturing precision
If existing purification methods are applied to industrial scale production, then product purity is improved, but production cost increases
Solution Approach 1:
The process uses disposable ultrafiltration and nanofiltration membranes that can be replaced rather than cleaned and sterilized repeatedly. This approach reduces the cost and complexity of maintaining purification systems at industrial scale, while still achieving high product purity through the sequential filtration and ion exchange steps.
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 efficient separation and purification of N-acetylglucosamine containing oligosaccharides, suitable for industrial applications, with high yield and purity, meeting regulatory requirements for food use.
Implementation Method 1
The method comprises the steps of: a) ultrafiltration of the fermentation broth and collecting the ultrafiltration permeate
Implementation Method 2
b) nanofiltration of the ultrafiltration permeate and collecting the nanofiltration retentate
Implementation Method 3
treatment with ion exchange resins, specifically strongly acidic cation and weakly basic anion exchange resins
Data Source
AI summary
The invention relates to a method for obtaining an N-acetylglucosamine containing neutral oligosaccharide from a fermentation broth, wherein said oligosaccharide is produced by culturing a genetically modified microorganism capable of producing said oligosaccharide from an internalized carbohydrate precursor, comprising the steps of: i) ultrafiltration (UF), preferably to separate biomass from the broth, ii) nanofiltration (NF), preferably to concentrate said oligosaccharide in the broth and/or reduce an inorganic salt content of the broth, and iii) treating the broth with an ion exchange resin, preferably to remove charged materials, and/or subjecting the broth to chromatography, preferably to remove hydrophobic impurities.