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Method for extracting EPA from nannochloropsis oculata by compound enzyme method

A technology of Nannochloropsis and compound enzymes, which is applied in the separation/purification of carboxylic acid compounds, oil/fat production, organic chemistry, etc. It can solve the problems of unsatisfactory extraction purity, further improvement of process conditions, and limited process extraction conditions and other problems, to achieve the effects of less pollution, increased fluidity, easy and safe operation

Active Publication Date: 2018-11-16
QINGDAO LANGYATAI GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is reported in the prior art to extract by aqueous enzymatic method, yet its extraction purity is not ideal, that is to say, the process extraction conditions in the prior art are limited, and the process conditions need to be further improved

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The first step is to centrifuge the Nannochloropsis fermented liquid at a speed of 3600rpm for 20 minutes to obtain the supernatant and algae mud, and collect the algae mud; select activated carbon in the suction filter barrel: diatomaceous earth = 1:1 to build a filter cake , after the supernatant is suction-filtered and adsorbed, it is used to configure a new medium, which can be recycled to achieve clean production. 5.0 kg can process 200 liters of supernatant;

[0025] The second step, adding pH to the algae mud is a phosphate buffer solution with a pH of 7.5, the ratio of material to water is 1:6 (g / mL), the amount of chitosanase added is 0.8% of the weight of the algae mud, and the amount of fructose enzyme added is 0.8% of the weight of the algae mud. 0.5% by weight, the amount of alkaline protease added is 1.8% by weight of algae mud, hydrolyzed for 0.8h at an enzymolysis temperature of 40°C to obtain an enzymolysis solution;

[0026] The third step is to centri...

Embodiment 2

[0030] The first step is to centrifuge the Nannochloropsis fermented liquid at a speed of 3600rpm for 20 minutes to obtain the supernatant and algae mud, and collect the algae mud; select activated carbon in the suction filter barrel: diatomaceous earth = 1:1 to build a filter cake , after the supernatant is suction-filtered and adsorbed, it is used to configure a new medium, which can be recycled to achieve clean production. 5.0 kg can process 200 liters of supernatant;

[0031] The second step is to add a phosphate buffer solution with a pH of 7.5 to the algae mud, the ratio of material to water is 1:6 (g / mL), the compound enzyme is chitosanase, fructose and alkaline protease, and the amount of chitosanase added 0.7% of the weight of the algae mud, the amount of fructose added is 0.6% of the weight of the algae mud, the amount of alkaline protease added is 1.9% of the weight of the algae mud, and the hydrolysis temperature is 40°C for 0.8 hours to obtain an enzymolyzed soluti...

Embodiment 3

[0036] The first step is to centrifuge the Nannochloropsis fermented liquid at a speed of 3600rpm for 20 minutes to obtain the supernatant and algae mud, and collect the algae mud; use activated carbon in the suction filter bucket: diatomaceous earth = 1:1 to build a filter cake , after the supernatant is suction-filtered and adsorbed, it is used to configure a new medium, which can be recycled to achieve clean production. 5.0 kg can process 200 liters of supernatant;

[0037] The second step is to add a phosphate buffer solution with a pH of 7.5 to the algae mud, the ratio of material to water is 1:6 (g / mL), the compound enzyme is hemicellulase, alkaline protease and trypsin, and the addition of hemicellulase The amount is 0.7% of the weight of the algae mud, the amount of alkaline protease added is 0.6% of the weight of the algae mud, the amount of trypsin added is 1.9% of the weight of the algae mud, and the hydrolysis temperature is 50°C for 0.6h to obtain the enzymolyzed s...

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Abstract

The invention discloses a method for extracting EPA from nannochloropsis oculata by a compound enzyme method. The method comprises the steps: subjecting nannochloropsis oculata fermentation liquor tocentrifugation, and collecting nannochloropsis oculata mud; adding a certain volume of buffer solution into the nannochloropsis oculata mud, then, adding a compound enzyme, and carrying out a reaction, so as to obtain an enzymolysis solution; subjecting the enzymolysis solution to centrifugation so as to obtain first free oil and an emulsion, subjecting the emulsion to further demulsified centrifugation so as to obtain second free oil, merging the first free oil and the second free oil, so as to obtain crude oil; and further purifying the obtained crude oil through a supercritical CO2 purification method, thereby obtaining the EPA. According to the method, the nannochloropsis oculata is treated by the compound enzyme, cell structures of microalgae are subjected to enzymolysis destruction under certain reaction conditions, meanwhile, the degradation action on complexes such as lipopolysaccharides and lipoproteins is strengthened, the fluidity of fat and oil is improved, and the fat andoil can be liberated. According to the method, the extraction purity of the EPA is 95% or more.

Description

technical field [0001] The invention relates to the technical field of biological extraction, in particular to a method for extracting EPA in Nannochloropsis by a compound enzyme method. Background technique [0002] EPA is Eicosapentaenoic acid (Eicosapentaenoic Acid), which has various physiological functions such as promoting cell metabolism, lowering cholesterol, relieving arthritis, preventing and treating cardiovascular and cerebrovascular diseases, and is widely used in human health care and emerging pharmaceutical industries. Natural EPA was mainly obtained from fish oil in the past, but the composition of fish oil is relatively complex, so it is difficult to extract high-purity EPA, which makes it difficult to exert its efficacy; and due to the reduction of fish resources worldwide, it is extracted from fish oil EPA has been unable to meet the increasing market demand. [0003] Nannochloropsis sp. is a marine unicellular microalgae with small algal cells with a dia...

Claims

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Application Information

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IPC IPC(8): C07C51/42C07C51/48C07C57/03C11B1/02C11B1/04C12P7/64
CPCC07C51/42C07C51/48C11B1/025C11B1/04C12P7/6427C07C57/03Y02P20/54
Inventor 李悦明许丽娜徐建春夏修峦徐炳政
Owner QINGDAO LANGYATAI GRP
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