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Method for preparing conjugated linoleic acid through immobilized lipase catalysis in fed-batch mode and application

A technology for immobilizing lipase and conjugated linoleic acid, which can be applied in the direction of fermentation, etc., can solve the problems of reduced reaction time, complex synthesis method and high cost

Active Publication Date: 2021-09-07
INNOBIO CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems of complex synthetic methods and high cost in the prior art, the solution adopted in the present invention is a tank-type stirring reaction, and at the same time, innovatively adopts the method of adding water in a vacuum flow to azeotrope out the alcohol and water, and add new water at the same time Continuing to respond quickly, reducing costs, reducing reaction time, increasing production capacity, simplifying post-processing, and facilitating subsequent production. The acid value of the obtained product is greater than or equal to 185, and the hydrolysis rate of the conjugated material is as high as 90-99%. At the same time , the content of conjugated linoleic acid in the product is 78-85%; the peroxide value is 0-1.0, and the anisidine value is 0-5

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] ——The process of adding water to vacuum flow

[0038] The safflower seed oil and ethanol are put into the reaction kettle with the molar ratio of 1:3.5, and 3% sodium hydroxide of the safflower seed oil mass is added, and the reaction is stirred and reacted at 80° C. for 3 hours after nitrogen replacement.

[0039]The reaction product is acidified until the pH is 7, stirred, left to stand, and the oil phase product is separated to obtain the safflower seed oil fatty acid ethyl ester product; the material obtained after the product falling film degassing is put into reaction with 3% potassium alcohol alcohol of the ethyl ester mass In the kettle, after nitrogen replacement, the mixture was stirred and reacted at 130°C for 3 hours to obtain the final product, ie, the ethyl ester conjugated compound.

[0040] Pack the immobilized enzyme into a pocket made with a 200-mesh screen. 20g of immobilized enzyme is loaded into each pocket, and the total enzyme amount is 4% of the...

Embodiment 2

[0049] 1. Put safflower seed oil and methanol into the reaction kettle together with 2.5% sodium hydroxide of the safflower seed oil mass according to the molar ratio of 1:3, and stir and react at 75°C for 4 hours after nitrogen replacement;

[0050] 2. Conjugation

[0051] The reaction product is acidified to a pH of 7 and stirred, left to stand, and the oil phase product is separated to obtain safflower seed oil fatty acid methyl ester; the material obtained after falling film degassing of the product and 3% potassium alkoxide by mass are put into the reactor, After nitrogen replacement, the mixture was stirred and reacted at 140° C. for 4 h to obtain the final product, namely the methyl ester conjugated compound.

[0052] 3. Enzymatic hydrolysis

[0053] Pack the immobilized enzyme into a pocket made with a 200-mesh screen. 25g of immobilized enzyme is loaded into each pocket, and the total enzyme amount is 4.5% of the amount of methyl ester conjugated material, which is ...

Embodiment 3

[0066] 1. Put safflower seed oil and propanol in a molar ratio of 1:4, add 2.5% sodium hydroxide of safflower seed oil mass into the reaction kettle, and stir and react at 70°C for 5 hours after nitrogen replacement;

[0067] 2. Conjugation

[0068] The reaction product is acidified to a pH of 6 and stirred, left to stand, and the oil phase product is separated to obtain propyl ester; the material obtained after falling film degassing of the product and 4% of the mass of potassium alkoxide are put into the reactor, and the nitrogen replacement is carried out at 160 The reaction was stirred at ℃ for 4 h to obtain the final product, namely the conjugated compound.

[0069] 3. Enzymatic hydrolysis

[0070] Pack the immobilized enzyme into a pocket made with a 100-mesh screen. 35g of immobilized enzyme is loaded into each pocket, and the total enzyme amount is 7% of the total mass of the ethyl ester conjugated material, which is put into the still after being sealed. Adjust the...

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Abstract

The invention discloses a method for preparing conjugated linoleic acid immobilized lipase catalysis in a fed-batch mode. The method comprises the following steps: after preparing an esterified conjugated material, filling the immobilized lipase into a pocket made of a screen, where the total enzyme amount is 1-10% of the esterified conjugated material; adjusting the pH value of the conjugated material to 5-7.5 by using an acid, uniformly mixing the conjugated material with water according to a mass ratio of 1: (0.5-10) to prepare a reaction solution, preheating, reducing pressure, steaming out the water and alcohol, feeding water at the same time so that the mass ratio of the added water to the discharged water is 1: (0.5-2), and reacting until the acid value is greater than or equal to 185; recovering the immobilized enzyme; and standing a separated oil-water mixture to separate water, dehydrating an oil layer, and carrying out molecular distillation. The peroxide value, the hydrolysis rate, the anisidine value and the like of a final product are obviously superior to those of the prior art. Meanwhile, the reaction is fast, the cost is low, the whole technological process is shortened and optimized, the productivity is increased, aftertreatment is simplified, and follow-up production is facilitated.

Description

technical field [0001] The invention relates to the technical field of immobilized lipase catalysis, in particular to a method and application for preparing conjugated linoleic acid catalyzed by immobilized lipase in a feeding mode. Background technique [0002] Conjugated linoleic acid (CLA for short) is a general term for several stereo and positional isomers of linoleic acid molecules, and is octadecadienoic acid containing conjugated double bonds. In the CLA series, c9, t11-CLA and t10, c12-CLA are the two main isomers with the most content and proven physiological activity. Natural CLA mainly exists in rumen animals, and the content is low, so humans basically cannot obtain it from nature. Conjugated linoleic acid has many effects, such as anti-arteriosclerosis, lowering blood lipids, regulating blood pressure and inhibiting some cancers. At the same time, its anti-oxidation, anti-fatigue, and immune-weight-loss-muscle-enhancing activities are being more and more deve...

Claims

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Application Information

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IPC IPC(8): C12P7/64
CPCC12P7/6427
Inventor 牛坤洪永德姜书华吴文忠
Owner INNOBIO CORP LTD
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