Immobilized Lipolytic Enzyme Regeneration via Solvent Washing
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Solution Overview
Problem
Conventional methods for regenerating immobilized lipolytic enzymes used in esterification reactions result in decreased diacylglycerol (DAG) purity due to residual enzyme activity, as the enzymes continue to catalyze interesterification reactions, leading to the production of triacylglycerol from 1,3-DAG, which lowers DAG purity.
Innovation Solution
The method involves washing the used immobilized lipolytic enzyme with n-hexane to reduce residual oil content, followed by treatment with an alkali solution to desorb the enzyme, and then re-adsorbing a fresh lipolytic enzyme onto the carrier, using an anion exchange resin as the immobilization carrier.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the used immobilized lipolytic enzyme is directly reused after simple filtration, then the productivity is maintained, but the DAG purity decreases due to residual enzyme activity causing interesterification reactions
Solution Approach 1:
The patent extracts and removes the residual lipolytic enzyme from the used immobilized carrier through washing with n-hexane and alkali treatment. This extraction process eliminates the harmful residual enzyme activity that would otherwise cause interesterification reactions and reduce DAG purity, while allowing the carrier itself to be reused for maintaining productivity
Solution Approach 2:
The patent changes the chemical environment parameters by treating the used immobilized enzyme with n-hexane followed by alkali solution. These parameter changes (solvent type, pH, ionic strength) alter the enzyme's adsorption state on the carrier, enabling complete removal of residual enzyme activity while preserving the carrier structure for reuse
2Adaptability or versatility
If alkali treatment is applied to remove all proteins from the immobilized enzyme, then the carrier can be reused, but the method is limited to specific immobilization carriers and cannot be universally applied
Solution Approach 1:
The patent creates a universal regeneration method that works across different immobilization carrier types (silica gel, activated carbon, ion exchange resins, etc.). The two-step process of n-hexane washing followed by alkali treatment serves multiple functions: removing residual oil, desorbing enzyme proteins, and preparing the carrier for reuse, making the method broadly applicable beyond single carrier types
Solution Approach 2:
The patent segments the enzyme removal process into two distinct stages: first washing with n-hexane to remove residual oil and loosely bound proteins, then treating with alkali solution to completely desorb remaining enzyme proteins. This segmentation makes the complex removal process more controllable and adaptable to different carrier types
3Manufacturing precision
If the residual oil content in the immobilized lipolytic enzyme is not reduced, then the process is simpler, but the enzyme activity cannot be effectively removed and DAG purity decreases
Solution Approach 1:
The patent performs preliminary washing with n-hexane before the main alkali treatment step. This preliminary action removes residual oil and loosely adsorbed proteins from the immobilized enzyme, reducing the burden on the subsequent alkali treatment and improving overall enzyme removal efficiency while maintaining manageable process complexity
Solution Approach 2:
The patent uses n-hexane as an intermediary substance that facilitates the removal process. The non-polar n-hexane effectively extracts residual oil and hydrophobic protein regions from the immobilized enzyme, preparing the carrier surface for more effective alkali treatment and enabling complete enzyme removal without excessive complexity
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 effectively regenerates the immobilized enzyme to match the activity of a new enzyme, allowing for the production of DAG-containing fat and oil with high DAG purity, achieving a DAG purity of 50% or more and reducing TAG content to 20% or less.
Implementation Method 1
mixing an immobilized lipolytic enzyme that has been used in an esterification reaction with n-hexane to wash the immobilized lipolytic enzyme so that a residual oil content in the immobilized lipolytic enzyme is reduced
Implementation Method 2
bringing the immobilized lipolytic enzyme into contact with an alkali solution; collecting the immobilization carrier
Implementation Method 3
adsorbing a lipolytic enzyme on the immobilization carrier
Data Source
AI summary
Provided is a method of producing an immobilized lipolytic enzyme which has almost the same ability as that of an unused immobilized enzyme by effectively utilizing an immobilization carrier in a used immobilized enzyme which has been used in an esterification reaction. The method of producing an immobilized lipolytic enzyme includes: mixing an immobilized lipolytic enzyme that has been used in an esterification reaction with a solvent to wash the immobilized lipolytic enzyme so that the residual oil content in the immobilized lipolytic enzyme is reduced to 50 parts by mass or less based on 100 parts by mass of an immobilization carrier; bringing the immobilized lipolytic enzyme into contact with an alkali solution; collecting the immobilization carrier; and adsorbing a lipolytic enzyme on the immobilization carrier.