Phosphatidylserine Purification Using Bivalent Metal Oxides

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Solution Overview

Problem

Current methods for producing and purifying phosphatidylserine (PS) face challenges such as high solvent requirements, costly production, and lower yields, particularly when using calcium salts in enzymatic conversion processes.

Innovation Solution

The use of bivalent metal oxides (BMO) in enzymatic transphosphatidylation reactions, which modify the reaction substrate and increase the conversion of phosphatidylcholine to PS, allowing for higher yields and more efficient purification in various solvents and media, including hydroalcoholic and aprotic mediums.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If calcium salts are used in enzymatic conversion processes, then the conversion of phosphatidylcholine to phosphatidylserine can be achieved, but the yield is lower and production costs are higher

Engineering Contradiction:
Improveyield of phosphatidylserine productionVSAvoidproduction cost
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent changes the chemical parameter of the reaction medium by using bivalent metal oxides (such as calcium oxide, magnesium oxide, zinc oxide) instead of calcium salts. This parameter change fundamentally alters the reaction conditions, leading to significantly improved conversion yields (up to 90%) and reduced production costs. The metal oxides modify the reaction substrate and enhance the enzymatic transphosphatidylation efficiency.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent employs bivalent metal oxides as a cost-effective alternative to expensive calcium salts and organic solvent systems. These metal oxides are inexpensive, readily available materials that can be easily removed from the reaction mixture, making the process economically viable while achieving high yields.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

2Manufacturing precision

If organic solvents are used for extraction and purification, then phosphatidylserine can be separated from the reaction mixture, but large quantities of solvent are required and elimination is difficult

Engineering Contradiction:
Improvepurity of phosphatidylserineVSAvoidsolvent quantity and elimination difficulty
Core Design Contradiction:
Manufacturing precisionVSLoss of substance

Solution Approach 1:

The patent extracts phosphatidylserine from the reaction mixture using water-miscible solvents (such as alcohols, ketones, or esters) instead of traditional organic solvents. These solvents can be easily removed by evaporation or dialysis, eliminating the difficulty of solvent elimination while achieving high purity product. The extraction process selectively separates PS from other reaction components.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent uses water-miscible solvents as intermediary substances that facilitate the extraction and purification of phosphatidylserine. These intermediary solvents bridge the aqueous reaction medium and the phospholipid product, enabling efficient separation while being easily removable without contaminating the final product.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Adaptability or versatility

If detergent/surfactant is used to disperse phospholipid in micellar form, then enzymatic reaction can occur in aqueous medium, but the process requires specific concentrations and conditions

Engineering Contradiction:
Improveability to conduct reaction in aqueous mediumVSAvoidprocess complexity with detergent/surfactant
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent replaces detergent/surfactant systems with bivalent metal oxides, which are simpler, cheaper, and easier to manage. The metal oxides do not require precise concentration control or special handling conditions, thereby reducing process complexity while maintaining the ability to conduct enzymatic reactions in aqueous medium.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent changes the fundamental parameter of the reaction system by eliminating the need for detergent/surfactant micellar structures. Instead, bivalent metal oxides create a different microenvironment that enables enzymatic activity in aqueous solution without requiring complex surfactant systems, thereby simplifying the overall process.

Inventive Principle:
Principle #35Parameter changes

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 approach significantly enhances the yield of PS production, achieving up to 90% conversion with reduced solvent usage and lower production costs, compared to methods using calcium salts, while maintaining high purity and efficiency.

Implementation Method 1

The use of bivalent metal oxides (BMO) in enzymatic transphosphatidylation reactions, which modify the reaction substrate and increase the conversion of phosphatidylcholine to PS

Methodology Applied
Scientific EffectEnzymatic transphosphatidylation: Enzyme

Implementation Method 2

The enzymatic conversion of phosphatidylcholine (PC) into PS by a transphosphatidylation reaction catalysed by the enzyme phospholipase D (PLD)

Methodology Applied
Scientific EffectCatalysis: Catalysis

Implementation Method 3

BMO drastically modifies the reaction substrate represented by the starting phosphatidylcholine, determining a change such that the action of enzyme PLD significantly increases in terms of final yield in PS

Methodology Applied
Scientific EffectChemical modification:

Implementation Method 4

with subsequent purification brought about mainly by extracting the PS with organic solvents

Methodology Applied
Scientific EffectSolvent extraction: Liquid-Liquid Extraction

Implementation Method 5

the PS, which has been deposited in the upper part of the reaction medium, is then isolated by separating and eliminating the subnatant

Methodology Applied
Scientific EffectPrecipitation: Precipitation

Data Source

PatentEP2431020B1Processes for the separation and purification of phosphatidylserine
Publication Date: 2014.07.16 FIDIA FARM SPA
  • EP2431020B1 patent drawingFigure 1
  • EP2431020B1 patent drawingFigure 2
  • EP2431020B1 patent drawingFigure 3

AI summary

The invention discloses advantageous processes for the separation and purification of phosphatidylserine (PS) from serine, PC and PLD remaining in a medium in which a transphosphatidylation reaction has taken place.