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
Engineering 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
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
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
2Manufacturing precision
If traditional isolation methods with multiple steps are used, then purity is improved, but productivity decreases due to time-consuming procedures
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
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
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
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
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
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
Implementation Method 2
adding one or more C 1 -C 4 alcohols to the solution
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
Data Source
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.