Method for Enriching Long-chain Polyunsaturated Fatty Acids Using Immobilized Thermomyces lanuginosus Lipase

A technology for immobilizing lipase and unsaturated fatty acids, which is used in the production of fat, fat oil/fat separation, fermentation, etc., can solve the problems of inability to meet the market demand for high-content ω-3 PUFAs, difficult hydrolysis reaction process, and low enrichment effect. , to achieve the effect of low price, reducing the use of organic reagents and simplifying the process

Active Publication Date: 2020-03-31
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantages of the lipase selective hydrolysis method are: the production cost is high due to the limitation of the hydrolytic enzyme source, the hydrolysis reaction process is difficult to control, the enrichment purity is low, and it cannot meet the market demand for high-content ω-3PUFAs
However, its enrichment effect on PUFAs is still not high, not as good as immobilized Mihei root hair enzyme lipase (Rhizo-mucor miehei, RM), PS (Pseudomonas sp.) and PF (Pseudomonas fluorescens)

Method used

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  • Method for Enriching Long-chain Polyunsaturated Fatty Acids Using Immobilized Thermomyces lanuginosus Lipase
  • Method for Enriching Long-chain Polyunsaturated Fatty Acids Using Immobilized Thermomyces lanuginosus Lipase
  • Method for Enriching Long-chain Polyunsaturated Fatty Acids Using Immobilized Thermomyces lanuginosus Lipase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0071]Enrichment and purification of PUFAs in embodiment 1DHA algae oil

[0072] (1) Alkali-catalyzed hydrolysis: First, dissolve 36.8g KOH in 70.4mL water and 422.4mL95% ethanol to prepare KOH alcohol-water solution, add 160g DHA algae oil under nitrogen protection, and reflux for 1 hour under magnetic stirring at 50°C. Hydrolyze fats. After the reaction, add 381mL of water, extract the unsaponifiable matter with 500mL×2 n-hexane, acidify the hydrated layer with 3N HCl to pH=1, extract the free fatty acid with 300mL×2 n-hexane, and then use anhydrous Na 2 SO 4 After drying, the solvent was removed by rotary evaporation to obtain free fatty acid.

[0073] (2) Weigh 9.57g of free fatty acid prepared in step (1) and 3g of glycerol (the molar ratio of free fatty acid and glycerol is about 1:1) into a 50mL reactor, add 10% immobilized TL lipase (to react Substrate (that is, the total mass of free fatty acid and glycerol) by mass), reacted at a stirring rate of 250rpm under vacu...

Embodiment 3

[0089] Enrichment and purification of PUFAs in embodiment 3 sardine oil

[0090] (1) Alkali-catalyzed hydrolysis: First, dissolve 18g KOH in 35mL water and 210mL 95% ethanol to prepare KOH alcohol-water solution, add 70g sardine oil under the protection of argon, and reflux for 0.5 hours under magnetic stirring at 60°C to hydrolyze the oil. After the reaction, add 200mL of water, extract the unsaponifiable matter with 200mL×2 n-hexane, and use 3N HNO 3 Acidify to pH=1, extract free fatty acids with 200mL×2 n-hexane, and then use anhydrous MgSO 4 After drying, the solvent was removed by rotary evaporation to obtain free fatty acid.

[0091] (2) Weigh 9.75g of 7 parts of free fatty acid prepared in step (1), and place them in a 50mL reactor with lauryl alcohol (the molar ratio of free acid and lauryl alcohol is shown in Table 4), and add 10% immobilized TL lipase (Based on the mass of the reaction substrate), the reaction was carried out at a stirring rate of 250 rpm for 12 ho...

Embodiment 4

[0095] Enrichment and purification of PUFAs in cod liver oil of embodiment 4

[0096] (1) Alkali-catalyzed hydrolysis: First, 26.3g NaOH was dissolved in 70.4mL water and 422.4mL95% ethanol to prepare KOH alcohol-water solution. Under nitrogen protection, 150g cod liver oil was added and refluxed for 1.5 hours under magnetic stirring at 65°C to make the oil hydrolysis. After the reaction, add 381mL of water, extract the unsaponifiable matter with 500mL×2 n-hexane, acidify the hydrated layer to pH=1 with 3N HCL, extract the free fatty acid with 300mL×2 n-hexane, and then use anhydrous CuSO 4 After drying, the solvent was removed by rotary evaporation to obtain free fatty acid.

[0097] (2) Weigh 7 parts of 9.70g of free fatty acid prepared in step (1) (produced by alkali-catalyzed hydrolysis of cod liver oil), and 3g of glycerin (the molar ratio of free acid to glycerin is about 1:1) in a 50mL reactor Inside, add immobilized TL lipase (based on the mass of the reaction substr...

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Abstract

The invention relates to a method for enriching long-chain polyunsaturated fatty acids through immobilized thermomyces lanuginosus. The method includes following steps: (1) carrying out an esterification reaction to a free fatty acid and an alcohol in the presence of the immobilized thermomyces lanuginosus in vacuum or a protective atmosphere; and (2) separating free long-chain polyunsaturated fatty acids out from a product after the reaction. By means of selectively catalytic esterification to fatty acids with different chain lengths during a catalytic esterification process of the fatty acid and the alcohol through immobilized lipases, a new method for enriching the long-chain polyunsaturated fatty acids from deep sea fish oil and microbial oil is achieved.

Description

technical field [0001] The present invention relates to a method for enriching long-chain polyunsaturated fatty acids using immobilized Thermomyces lanuginosus lipase, which uses immobilized lipase to catalyze the esterification of fatty acids and alcohols The selective catalytic esterification of fatty acids with different chain lengths has realized a new method for enriching long-chain polyunsaturated fatty acids from deep-sea fish oil or microbial oil. [0002] technical background [0003] In recent years, long-chain polyunsaturated fatty acids (PUFAs) such as EPA, DHA, DPA, and ARA have been research hotspots because of their unique physiological functions. Due to the low content of naturally occurring PUFAs, in order to improve the medical and health value of PUFAs, more and more research work has been devoted to the enrichment and purification of PUFAs. [0004] At present, the separation and purification methods of PUFAs mainly adopt traditional chemical methods, suc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C11B7/00C12P7/64
Inventor 杨武林郑妍辛本荣杨天奎徐学兵
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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