Recombinant Lipase rCRL1 for Cis-Unsaturated Fatty Acid Production

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Solution Overview

Problem

Industrial production of cis-unsaturated fatty acids often results in the conversion of cis-unsaturated fatty acids to trans-unsaturated fatty acids due to high-temperature and high-pressure treatments, leading to health concerns and inefficiencies, and existing enzyme applications face issues with non-reproducibility and high costs.

Innovation Solution

A method using recombinant Candida rugosa lipase 1 (rCRL1) for low-temperature hydrolysis of oil-water mixtures, involving emulsification and controlled temperature and filtration processes to produce cis-unsaturated fatty acids without the need for organic solvents, ensuring stable enzyme quality and reduced production costs.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If high-temperature and high-pressure treatment is used for industrial production of edible oil, then production efficiency is improved, but cis-unsaturated fatty acids are converted to trans-unsaturated fatty acids

Engineering Contradiction:

Improveproduction efficiencyVSAvoidtrans-unsaturated fatty acid conversion

Core Design Contradiction:

ProductivityVSObject-generated harmful factors

Solution Approach 1:

The patent changes the temperature parameter from high-temperature (210-265°C) to low-temperature (30-50°C) treatment, which prevents the conversion of cis-unsaturated fatty acids to trans-unsaturated fatty acids while maintaining production efficiency through enzymatic catalysis

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent replaces the mechanical/thermal system (high-temperature and high-pressure treatment) with a biochemical system (enzymatic hydrolysis using lipase), which achieves fat hydrolysis at low temperatures without causing unwanted isomerization reactions

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Adaptability or versatility

If commercial CRL preparations containing multiple isoenzymes are used for hydrolysis, then enzyme versatility is improved, but reaction reproducibility deteriorates

Engineering Contradiction:

Improveenzyme versatilityVSAvoidreaction reproducibility

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent extracts and uses only the most effective isoenzyme (CRL1) from the mixture of multiple isoenzymes in commercial CRL preparations, eliminating the variability introduced by other isoenzymes while maintaining catalytic effectiveness

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent applies the principle of local quality by selecting a specific isoenzyme (CRL1) with optimal properties for the intended application, rather than using a general mixture of isoenzymes, thereby achieving both effectiveness and reproducibility

Inventive Principle:

Principle #3Local quality

3Duration of action of stationary object

If immobilized enzymes and organic solvents are used in enzyme reactions, then enzyme reusability is improved, but production complexity and environmental impact worsen

Engineering Contradiction:

Improveenzyme reusabilityVSAvoidproduction complexity

Core Design Contradiction:

Duration of action of stationary objectVSDevice complexity

Solution Approach 1:

The patent uses free (non-immobilized) lipase that can be produced cheaply through fermentation and used in a single batch process, eliminating the need for complex immobilization techniques and organic solvents while maintaining cost-effectiveness

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent employs fermentation broth containing lipase that can be directly used for hydrolysis without additional purification or immobilization steps, allowing the system to serve itself by utilizing the natural state of the enzyme produced through fermentation

Inventive Principle:

Principle #25Self-service

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 effectively produces cis-unsaturated fatty acids with high yields and retention of nutrients, avoiding the conversion to trans-unsaturated fatty acids and reducing enzyme costs, while maintaining operational efficiency and environmental sustainability.

Implementation Method 1

hydrolyzing the emulsified oil-water mixture to generate fatty acid

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

recombinant Candida rugosa lipase 1 (rCRL1) for low-temperature hydrolysis

Methodology Applied

Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 3

emulsifying the oil-water mixture

Methodology Applied

Scientific EffectEmulsification: Emulsion

Implementation Method 4

performing a cooling and filtering step to obtain cis-unsaturated fatty acid

Methodology Applied

Scientific EffectFiltration: Filter (physical)

Data Source

PatentUS11685939B2Method for producing cis-unsaturated fatty acid by recombinant <i>Candida rugosa </i>lipase 1 (rCRL1)

Publication Date: 2023.06.27 CHANT OIL

AI summary

A method for producing cis-unsaturated fatty acid includes the operations below. (i) An oil-water mixture is provided, wherein the oil-water mixture includes 1 to 10 parts by weight of oil and 1 part by weight of water. (ii) 0.002 to 0.5 parts by weight of a recombinant Candida rugosa lipase 1 (rCRL1) is added into the oil-water mixture. (iii) The oil-water mixture is emulsified. (iv) The emulsified oil-water mixture is hydrolyzed and fatty acid is generated. (v) Oil-water is separated at a temperature of 55° C. to 65° C. and an oil phase layer is extracted. (vi) The cooling and filtering step is performed to obtain cis-unsaturated fatty acid.