Phosphatidylserine Production via Enzyme Inactivation
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Solution Overview
Problem
There is a lack of an economically viable and stable method for producing phosphatidylserine with a high content of polyunsaturated fatty acids, particularly DHA-bonded phosphatidylserine, from a safe starting material, due to issues with phospholipase A2 residue and hydrolysis during synthesis processes.
Innovation Solution
A method involving esterification of polyunsaturated fatty acids with lysophospholipid using phospholipase A2, followed by adjusting enzyme activity to 10 U/g or less, and performing a base exchange reaction with serine in the presence of phospholipase D to form phosphatidylserine, while removing or inactivating phospholipase A2 to prevent detachment of the fatty acid.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If phospholipase A2 is used to hydrolyze fatty acid at 2-position of phospholipid, then lysophospholipid is obtained, but phospholipase A2 remains in the product causing DHA detachment during storage
Solution Approach 1:
The patent applies preliminary action by inactivating phospholipase A2 through specific treatment (heating at 60-80°C for 30 minutes or adding calcium ions) before the DHA bonding step. This preliminary inactivation prevents the enzyme from causing DHA detachment during subsequent storage and processing, thereby ensuring product stability while maintaining ease of manufacture.
2Productivity
If phospholipase D is used for serylation of phospholipid, then phosphatidylserine is formed, but hydrolysis at 2-position occurs requiring water
Solution Approach 1:
The patent applies preliminary anti-action by pre-inactivating phospholipase A2 and controlling reaction conditions (pH 7.5-8.5, temperature 37-42°C) before initiating the phospholipase D-mediated serylation. This prevents hydrolysis at the 2-position from occurring as a harmful side reaction, allowing efficient phosphatidylserine synthesis without compromising product integrity.
3Reliability
If DHA-bonded phosphatidylcholine is extracted from fish tissues, then DHA-bonded phosphatidylserine can be synthesized, but starting material is expensive and supply is unstable
Solution Approach 1:
The patent replaces expensive, supply-unstable fish tissue-derived phospholipids with cheaper, readily available plant-derived phospholipids (soybean lecithin, sunflower lecithin, or rapeseed lecithin). Although plant phospholipids have different fatty acid profiles, the process achieves the desired DHA-bonded phosphatidylserine product through enzymatic modification, dramatically reducing production costs and ensuring stable supply while maintaining product quality.
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 allows for the stable and cost-effective production of phospholipid compositions with 10% or more phosphatidylserine and 10-40% polyunsaturated fatty acids, ensuring effective bonding and stability of DHA-bonded phosphatidylserine.
Implementation Method 1
performing an esterification reaction of a polyunsaturated fatty acid with lysophospholipid using phospholipase A2 to obtain phospholipid
Implementation Method 2
using phospholipase A2 (PLA2) to obtain phospholipid
Implementation Method 3
performing a base exchange reaction of a mixture including the phospholipid and serine in the presence of phospholipase D (PLD) to form a phospholipid-containing composition which includes phosphatidylserine
Data Source
AI summary
A method for producing a phospholipid-containing composition which includes 10% by weight or more of phosphatidylserine based on the whole phospholipid-containing composition, a content of a polyunsaturated fatty acid being from 10 to 40% by weight based on the total amount of constituent fatty acids, the method including the following steps (1) and (2) in this order, and the following steps (3) and (4) in this order inexpensively and stably supplies a phospholipid-containing composition which includes phosphatidylserine to which a large amount of the polyunsaturated fatty acid is bonded at the 2-position thereof. Step (1): performing an esterification reaction of a polyunsaturated fatty acid with lysophospholipid using phospholipase A2 (PLA2) to obtain phospholipid. Step (2): adjusting an activity of PLA2 in the phospholipid to 10 U/g (phospholipid) or less after the step (1). Step (3): performing a base exchange reaction of a mixture including the phospholipid and serine in the presence of phospholipase D (PLD) to form a phospholipid-containing composition which includes phosphatidylserine. Step (4): separating the composition.