Enzyme Removal via Organic Solvent Mixture

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Solution Overview

Problem

Existing methods for deactivating enzymes used in phospholipid hydrolysis and base exchange reactions, such as heating, are inadequate as phospholipases exhibit strong thermal resistance and can lead to quality issues and safety concerns due to residual enzyme activity and protein presence.

Innovation Solution

A method involving a solvent mixture of water and an organic solvent containing an inorganic metal salt is used to efficiently remove residual enzyme activity from liquid enzyme reaction mixtures, effectively reducing enzyme and protein levels.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If heating treatment is used to deactivate phospholipase, then enzyme activity is reduced, but phospholipids and free fatty acids deteriorate and strong thermal resistance remains

Engineering Contradiction:
Improveenzyme deactivation efficiencyVSAvoidphospholipid deterioration
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent changes the deactivation parameter from thermal (heating) to chemical (organic solvent treatment). By using organic solvents such as acetone, ethyl acetate, or n-hexane, the patent achieves enzyme deactivation without the thermal damage that causes phospholipid deterioration and free fatty acid degradation.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent replaces the thermal deactivation mechanism with a chemical solvent-based mechanism. Instead of using heat energy to denature the enzyme, the patent uses organic solvent molecules to interact with and deactivate the phospholipase, avoiding the harmful thermal effects while achieving reliable enzyme deactivation.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Reliability

If heating at high temperature is used to deactivate phospholipase, then enzyme activity is completely reduced, but the process becomes problematic due to phospholipid deterioration

Engineering Contradiction:
Improveenzyme deactivation completenessVSAvoidproduct quality
Core Design Contradiction:
ReliabilityVSManufacturing precision

Solution Approach 1:

The patent changes the deactivation parameter from thermal (heating at 100°C or more) to chemical (organic solvent treatment). By using organic solvents such as acetone, ethyl acetate, or n-hexane, the patent achieves complete enzyme deactivation without the thermal damage that causes phospholipid deterioration and free fatty acid degradation.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent replaces the thermal deactivation mechanism with a chemical solvent-based mechanism. Instead of using heat energy to denature the enzyme, the patent uses organic solvent molecules to interact with and deactivate the phospholipase, avoiding the harmful thermal effects while achieving reliable enzyme deactivation.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

3Reliability

If protease treatment is used to deactivate phospholipase, then enzyme activity is reduced, but peptides and proteases remain causing allergy risk

Engineering Contradiction:
Improveenzyme deactivationVSAvoidallergy risk from peptides
Core Design Contradiction:
ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent replaces the protease-based biological deactivation mechanism with a chemical solvent-based mechanism. By using organic solvents, the patent directly deactivates the phospholipase without requiring proteolytic breakdown, thereby avoiding the generation of peptide fragments that could trigger allergic reactions.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent uses organic solvent molecules as intermediaries to deactivate the phospholipase. The organic solvent acts as a mediator that interacts with the enzyme structure to cause deactivation without requiring proteolytic cleavage, thus avoiding the production of potentially allergenic peptide fragments.

Inventive Principle:
Principle #24Intermediary (Mediator)

4Reliability

If filtration and adsorption processes are added to remove proteins, then protein removal is achieved, but the process becomes more complicated

Engineering Contradiction:
Improveprotein removal efficiencyVSAvoidprocess complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent replaces complex mechanical separation processes (filtration and adsorption) with a simpler chemical treatment approach. By using organic solvent treatment, the patent achieves both enzyme deactivation and protein removal in a single step, eliminating the need for additional filtration and adsorption equipment and processes.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent merges the enzyme deactivation function and the protein removal function into a single organic solvent treatment step. This consolidation eliminates the need for separate filtration and adsorption processes, thereby simplifying the overall process while maintaining effective protein removal.

Inventive Principle:
Principle #5Merging (Combining)

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 achieves significant reduction in enzyme activity and protein levels, improving product quality and safety by avoiding the limitations of traditional deactivation methods, such as heating, and ensuring the removal of enzymes without complicating the process.

Implementation Method 1

treating the liquid enzyme reaction mixture with a solvent mixture of water and an organic solvent, wherein the solvent mixture contains an inorganic metal salt, to remove the enzyme

Methodology Applied
Scientific EffectExtraction: Liquid-Liquid Extraction

Implementation Method 2

efficiently remove residual enzyme activity from liquid enzyme reaction mixtures, effectively reducing enzyme and protein levels

Methodology Applied
Scientific EffectDenaturation:

Data Source

PatentUS7709238B2Method for removing enzyme and method of base exchange or hydrolysis of phospholipid using the same
Publication Date: 2010.05.04 HAYASHIBARA CO LTD

AI summary

A method of removing an enzyme from a liquid enzyme reaction mixture used in a hydrolysis reaction or a base exchange reaction of a phospholipid is provided. The method includes the step of treating the liquid enzyme reaction mixture with a solvent mixture of water and an organic solvent, wherein the solvent mixture includes an inorganic metal salt, to remove the enzyme. Enzymes included in the reaction product can be easily removed without a treatment such as heating, and thus it becomes possible to easily produce various phospholipids that have a reduced risk of inducing an allergy, that retain a high quality and that have excellent storage stability.