Selective Crystallization of 2'-O-Fucosyllactose Using Alcohols

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Solution Overview

Problem

Current methods for isolating 2'-O-fucosyllactose (2'-FL) from biotechnological production, particularly fermentation broths, are inefficient due to the need for costly and time-consuming chromatographic separations, and existing crystallization methods are hindered by byproducts in the aqueous culture medium.

Innovation Solution

Selective crystallization of 2'-FL from an aqueous solution containing other fucosylated carbohydrates by adding C1-C4 alcohols, such as methanol, which allows for high-purity 2'-FL crystallization even in the presence of contaminants like 2',3-di-O-fucosyllactose, enabling large-scale production with high yields.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If chromatographic separation is used to isolate 2'-FL from fermentation broth, then purity of 2'-FL is improved, but cost and processing time increase significantly

Engineering Contradiction:
Improvepurity of 2'-FLVSAvoidcomplexity of separation process
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent changes the physical-chemical parameters of the fermentation broth by adjusting pH to specific ranges (4.5-5.5 or 6.0-7.5) and controlling temperature (0-25°C), which causes selective crystallization of 2'-FL. This parameter-based separation replaces complex chromatographic methods with simple crystallization, achieving high purity without expensive equipment or lengthy procedures

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent utilizes the phase transition of 2'-FL from dissolved state to crystalline state through controlled cooling and pH adjustment. By manipulating solubility parameters and inducing crystallization, 2'-FL separates from the aqueous phase as pure crystals, eliminating the need for chromatographic separation while achieving >95% purity

Inventive Principle:
Principle #36Phase transitions

2Manufacturing precision

If traditional isolation methods with multiple steps are used, then purity is improved, but productivity decreases due to time-consuming procedures

Engineering Contradiction:
Improvepurity of 2'-FLVSAvoidproduction efficiency
Core Design Contradiction:
Manufacturing precisionVSProductivity

Solution Approach 1:

The patent extracts 2'-FL from the fermentation broth through selective crystallization induced by pH adjustment and controlled cooling. This single extraction step replaces multiple sequential isolation steps (centrifugation, filtration, chromatography), dramatically reducing processing time while maintaining high purity. The method achieves both extraction and purification in one operation

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent combines multiple functions (separation, purification, concentration) into a single crystallization step. By adjusting pH and temperature simultaneously, the process achieves separation of 2'-FL from contaminants, concentration of the product, and purification all in one operation, thereby improving productivity without sacrificing purity

Inventive Principle:
Principle #5Merging (Combining)

3Ease of manufacture

If crystallization is used to isolate 2'-FL, then cost and simplicity are improved, but presence of byproducts in culture medium reduces effectiveness

Engineering Contradiction:
Improvesimplicity of isolation methodVSAvoidpurity of 2'-FL
Core Design Contradiction:
Ease of manufactureVSManufacturing precision

Solution Approach 1:

The patent performs preliminary pH adjustment of the fermentation broth to specific ranges (4.5-5.5 or 6.0-7.5) before crystallization. This preliminary action creates optimal conditions for selective 2'-FL crystallization while keeping other byproducts in solution. The pre-adjustment of pH ensures that when cooling occurs, only 2'-FL crystallizes, leaving contaminants in the aqueous phase

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent uses pH adjustment as an intermediary mechanism to enable selective crystallization. By controlling pH as an intermediate parameter, the process distinguishes 2'-FL from other carbohydrates and byproducts in the fermentation broth. This intermediary control allows simple crystallization to achieve high purity that would otherwise require complex separation methods

Inventive Principle:
Principle #24Intermediary (Mediator)

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 method achieves high-purity 2'-FL crystallization in a single step, facilitating large-scale production with wide ranges of contaminant concentrations, reducing costs and processing time compared to traditional chromatographic methods.

Implementation Method 1

Selective crystallization of 2'-FL from an aqueous solution containing other fucosylated carbohydrates by adding C1-C4 alcohols

Methodology Applied
Scientific EffectCrystallization: Crystallisation

Implementation Method 2

adding one or more C 1 -C 4 alcohols to the solution

Methodology Applied
Scientific EffectSolubility change: Solvation

Implementation Method 3

crystalline 2'-FL displays X-ray powder diffraction reflections, based on a measurement using CuKα radiation, at 16.98±0.20, 13.65±0.20 and 18.32±0.20 2Θ angles

Methodology Applied
Scientific EffectX-ray diffraction: X-Ray

Data Source

PatentEP3154995B1Separation of 2'-o-fucosyllactose from fermentation broth
Publication Date: 2023.11.01 GLYCOM AS

AI summary

The invention relates to a method for selective crystallization of 2'-O-fucosyllactose (2'-FL) from an aqueous solution comprising 2'-FL and a fucosylated carbohydrate other than 2'-FL, preferably 2',3-di-O-fucosyllactose (DFL), by adding one or more C1-C4 alcohols, preferably methanol, to the solution.