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Method for preparing glyceride rich in algal oil n-3 polyunsaturated fatty acid through enzyme process

An unsaturated fatty acid, algal oil-rich technology, applied in fermentation, chemical recovery, etc., can solve the problems of high reaction temperature, long reaction time, and low enrichment efficiency.

Active Publication Date: 2012-09-12
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzymatic selective hydrolysis process is relatively simple, and the enrichment efficiency is not high. The content of n-3PUFA in glycerides can only reach about 50%; high, and the enzymatic selective alcoholysis products are mixed glycerides; the substrates of enzymatic esterification synthesis reaction fatty acids and glycerol are dispersed in a two-phase system, which is not conducive to the mass transfer of the reaction; Saturated fatty acids, such as Chinese patents CN101161819A and CN101348807A, disclose the use of fish oil n-3PUFA concentrate and glycerol to enrich DHA and EPA through esterification
However, both of the above two patents use fish oil as the raw material, and the n-3PUFA concentrate is prepared by chemical hydrolysis in the whole process. The reaction temperature of the chemical method is relatively high, and a large amount of alkali is used as a catalyst. -3PUFA will have a certain destruction, and produce a large amount of pollutants such as alkali waste water; Synthetic reaction all is to carry out esterification reaction with glycerin and the n-3PUFA concentrate that chemical method prepares as substrate, because the two are mutually incompatible The two substrates are compatible, so the mass transfer of the reaction will be affected, resulting in a longer reaction time

Method used

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  • Method for preparing glyceride rich in algal oil n-3 polyunsaturated fatty acid through enzyme process
  • Method for preparing glyceride rich in algal oil n-3 polyunsaturated fatty acid through enzyme process
  • Method for preparing glyceride rich in algal oil n-3 polyunsaturated fatty acid through enzyme process

Examples

Experimental program
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Effect test

example 1

[0062] (1) Enzymatic partial hydrolysis of algae oil

[0063] Weigh 10 g of algae oil sample, add pH 8.0 phosphate buffer according to the mass ratio of algae oil to buffer of 1:0.6, add Candida sp.99-125 lipase enzyme according to the amount of 1500U / g algae oil powder, add cyclodextrin in an amount twice the mass of enzyme powder, fill nitrogen as an inert gas for sealing protection, hydrolyze at 50°C until the hydrolysis rate reaches 60%, stop the reaction, remove enzyme powder by suction filtration, and filtrate Neutralize with KOH, add n-hexane to extract partially hydrolyzed glycerides, and wash the extract with deionized water until neutral, and pass through anhydrous Na 2 SO 4 Drying is carried out, and the solvent is distilled off under reduced pressure to obtain partially hydrolyzed glycerides primarily enriched in n-3 polyunsaturated fatty acids as the main product. The hydration layer was acidified with 2mol / L HCl, and then the free fatty acids were extracted wit...

Embodiment 2

[0071] Embodiment 2 is different from embodiment 1 in that:

[0072] In step (1), the mass ratio of algae oil to buffer is 1:1, lipase enzyme powder is added according to the amount of 600U / g algae oil, and cyclodextrin is added according to the amount of 0.5 times the mass of enzyme powder; Under hydrolysis, after the hydrolysis rate reaches 40%, stop the reaction.

[0073] In step (2), according to the mass ratio of algae oil and buffer solution, add the amount containing 5mMCaCl 2 phosphate buffer solution, add lipase enzyme powder according to the amount of 600U / g algae oil, add cyclodextrin according to the amount of 0.5 times the mass of enzyme powder; carry out hydrolysis reaction at 35 ° C, after the hydrolysis rate reaches 20%, according to The amount of 0.5wt% algae oil is supplemented with Ca(OH) 2 , The total hydrolysis time is 48h.

[0074] In step (3), the ratio of fatty acid: urea: ethanol is 1:2:5 (w / w / v), blending at 60°C for 60 minutes, and inclusion at 4°...

Embodiment 3

[0078] Embodiment 3 is different from embodiment 1 in that:

[0079] In step (1), the mass ratio of algae oil to buffer is 1:0.8, lipase enzyme powder is added according to the amount of 1200U / g algae oil, and cyclodextrin is added according to an amount 1.5 times the mass of the enzyme powder; Under hydrolysis, after the hydrolysis rate reaches 59%, stop the reaction.

[0080] In step (2), according to the mass ratio of algae oil and buffer solution, add the amount containing 8mMCaCl 2 Phosphate buffer solution, add lipase enzyme powder according to the amount of 1200U / g algae oil, add cyclodextrin according to the amount of 1.5 times the mass of enzyme powder; carry out hydrolysis reaction under the condition of 40 ℃.

[0081] In step (3), the ratio of fatty acid: urea: ethanol is 1: 3.5: 9 (w / w / v), and they are blended at 70°C.

[0082] In step (4), the molar ratio of partially hydrolyzed glyceride to n-3 polyunsaturated fatty acid concentrate is 1:8, and immobilized lipa...

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Abstract

The invention relates to a method for preparing glyceride rich in algal oil n-3 polyunsaturated fatty acid through the enzyme process. The method comprises the following steps: taking microbial algal oil as raw material, carrying out hydrolysis of part or all of algal oil respectively through controlling the hydrolysis rate under the catalysis of lipases to obtain glyceride partially hydrolyzed and free fatty acid; enabling the free fatty acid to enrich n-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) through the urea adduct method to obtain an n-3 polyunsaturated fatty acid condensate; and taking the n-3 polyunsaturated fatty acid condensate and glyceride partially hydrolyzed as substrates, taking immobilized lipases as catalysts, and efficiently synthetizing glyceride rich in algal oil n-3 polyunsaturated fatty acid in an organic phase. The invention adopts a complete biological enzymatic enrichment process, the raw material is clean and safe, the process route is scientific and reasonable, the catalytic activity of enzyme is high, the catalyst can be recycled, the cost is low, the reaction condition is mild, the energy consumption is low, the environment is friendly, and the market prospect is wide.

Description

technical field [0001] The invention belongs to the technical field of biosynthesis and enzyme catalysis of polyunsaturated fatty acid glycerides, and relates to a method for enzymatically preparing glycerides rich in algae oil n-3 polyunsaturated fatty acids. A method for synthesizing glycerides with high n-3 polyunsaturated fatty acid content catalyzed by biological enzyme method. Background technique [0002] n-3 polyunsaturated fatty acids (n-3PUFA) play a pivotal role in human health and have received more and more attention in recent years. Its characteristic fatty acids DHA (cis-4, 7, 10, 13, 16, 19-Docosahexaenoic Acid, docosahexaenoic acid) and EPA (cis-5, 8, 11, 14, 17-Eicosapentaenoic Acid, 20 Carbapentaenoic acid) is favored by people because of its unique physiological functions. Among them, EPA has the effects of improving blood circulation, regulating blood lipids, lowering blood pressure, softening blood vessels, and anti-inflammation. It can be used to tre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/64
CPCY02P20/584
Inventor 谭天伟杨晓丹陈必强陶一峰崔彩霞钱轶欢
Owner BEIJING UNIV OF CHEM TECH
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