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A method for preparing conjugated linoleic acid by coupling method using immobilized lipase as catalyst

A technology of immobilized lipase and conjugated linoleic acid, which is applied in the direction of fermentation, etc., can solve the problems of unsatisfactory industrial production, long reaction time, and low product quality, so as to improve the reuse rate, shorten the enzyme reaction time, and improve The effect of product quality

Active Publication Date: 2016-01-06
INNOBIO CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Through the comparative analysis of the above patents and articles, it can be seen that the preparation of conjugated linoleic acid by chemical method is cumbersome, the product quality is low, and it has adverse effects on the environment.
The reaction efficiency of using immobilized lipase to hydrolyze oil is not high, which is mainly reflected in the conversion rate and reaction time. Generally, the reaction time is relatively long, which cannot meet the needs of industrial production.
The enzyme-catalyzed substrate used in the above patents - methyl conjugated linoleate is basically a product after acidification, water washing and distillation. This kind of treatment process is complicated, and the content of non-essential isomers is likely to increase during the distillation process. and higher cost
The above patents basically did not explain the issue of the recycling rate of immobilized lipase, which is a key point that directly restricts large-scale industrial production, and also did not clarify the quality of the prepared products

Method used

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  • A method for preparing conjugated linoleic acid by coupling method using immobilized lipase as catalyst
  • A method for preparing conjugated linoleic acid by coupling method using immobilized lipase as catalyst
  • A method for preparing conjugated linoleic acid by coupling method using immobilized lipase as catalyst

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] (1) Take 100g of methyl ester conjugated material, add an appropriate amount of 50% sulfuric acid / water at 50 degrees to neutralize to pH=8, and then continue to acidify for 1 hour to obtain acidified methyl ester conjugated material, the methyl ester conjugated The content of methyl conjugated linoleate in the chemical material is 80wt%;

[0048] (2) Fill 5g of immobilized lipase in the enzyme reaction column, and keep the temperature of the enzyme reaction column at 45°C;

[0049] (3) Prepare the acidified methyl ester conjugated material obtained in step (1) and water at a mass ratio of 1:0.5 to form a reaction solution, stir the reaction solution evenly to obtain an emulsified system, and preheat it to 40°C, pass through a constant flow The pump is pumped into the enzyme reaction column prepared in step (2), the stirring speed is 100rpm / min, the flow rate is 0.5ml / min, and the reaction is continued for 4 hours to obtain the reaction product. Continue to use 50% sulf...

Embodiment 2

[0051] (1) Add 50% citric acid aqueous solution to 100 g of methyl ester conjugated material, neutralize to pH = 8, and then continue to acidify for 1 hour to obtain acidified methyl ester conjugated material. The methyl ester conjugated material is co- Conjugated linoleic acid methyl ester content is 80wt%;

[0052] (2) Fill 5g of immobilized lipase in the enzyme reaction column, and keep the temperature of the enzyme reaction column at 40°C;

[0053] (3) Prepare the acidified methyl ester conjugated material obtained in step (1) and water at a mass ratio of 1:0.5 to form a reaction solution, stir the reaction solution evenly, and preheat it to 40°C, and pump it into the step through a constant flow pump (2) In the prepared enzyme reaction column, the stirring speed is 100rpm / min, the flow rate is 0.5ml / min, and the reaction is continued for 4h, and the reaction product obtained is acidified to pH7. 0, and then centrifuged to obtain the oil phase, which is the final product....

Embodiment 3

[0055] Obtain the acidified methyl ester conjugated material according to the method of implementing the step (1) of 2, wherein the organic acid used for acidification is 50% aqueous solution of wolfberry acid, and the content of methyl conjugated linoleate in the methyl ester conjugated material is 80wt%.

[0056] The enzyme reaction column filled with immobilized lipase was prepared according to the method in Example 2.

[0057] The prepared acidified methyl ester conjugated material and water were formulated into a reaction solution at a mass ratio of 1:0.5, the reaction solution was stirred evenly, and preheated to 40°C, and pumped into the enzyme reaction column through a constant flow pump at a stirring speed of 100rpm / min, flow rate 0.5ml / min, continuous reaction for 4 hours, the obtained reaction product was acidified to about pH7.0 with 50% aqueous solution of wolfberry acid at 80°C, and then centrifuged to obtain the oil phase, which is the final product , determine...

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Abstract

The invention provides a method for preparing conjugated linoleic acid through a coupling method using an immobilized lipase as a catalyst. The method comprises the steps of neutralizing a methyl or ethyl conjugated material to reach a pH value of 7.5 to 9 by using an organic acid, forming a uniform emulsification system with a small amount of water, and preparing the conjugated linoleic acid through hydrolysis by using the immobilized lipase as a catalytic substrate, wherein the conjugated material is safflower seed oil which contains more than 80% of linoleic acid, and is prepared through base catalysis isomerization. By adopting the emulsification system, the reaction time is greatly shortened, the hydrolysis efficiency of the methyl / ethyl conjugated material is improved, the dosage of the water is reduced, and the industrial application is facilitated. According to the method, other emulsifiers are not introduced in the preparation steps, the difficulties in subsequent separation and purification and the influences of the residual emulsifier on the quality can not be brought, and the product loss due to the separation and purification can not be caused. Meanwhile, the immobilized lipase used in the catalytic process can be recycled for 100 times repeatedly, and the reduction of the enzyme activity is less than or equal to 5%, so that the production cost is greatly reduced.

Description

technical field [0001] The invention relates to a method for preparing conjugated linoleic acid, in particular to a method for preparing conjugated linoleic acid through a coupling method using immobilized lipase as a catalyst. Background technique [0002] Conjugated linoleic acid (CLA) is a general term for several stereo and positional isomers of linoleic acid (Linoleic acid, LA) molecules, and is a series of octadecadienoic acids containing conjugated double bonds. The double bond of CLA mainly has four positions on the carbon chain: 8,10-, 9,11-, 10,12- and 11,13-, and because the carbon atoms at both ends of the conjugated double bond have cis Formula (cis) and trans (trans) two geometric configurations, that is, each positional isomerism has four geometric isomers of cis-cis, cis-trans, trans-cis, and trans-trans, so CLA isomerism There are many types of bodies. Among them, c9, t11-CLA and t10, c12-CLA are the two major isomers with the most content and proven physi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/64
Inventor 杨大苹吴文忠洪永德徐维锋付亮亮
Owner INNOBIO CORP LTD