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Method for extracting nannochloropsis oculata EPA polar lipid under assistance of enzyme treatment

A Nannochloropsis, auxiliary extraction technology, applied in the direction of fat production, fat oil/fat production, fat oil/fat refining, etc., can solve the problem of not being able to directly obtain pure EPA polar lipids and increase the extraction rate of Nannochloropsis polar lipids And other issues

Active Publication Date: 2021-05-14
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies in the prior art, the present invention provides a method for assisting the extraction of Nannochloropsis EPA polar lipids by enzyme treatment, by reselecting the enzyme species , and redesigned and optimized the enzymatic hydrolysis process and the overall extraction method steps, and finally broke the influence of the traditional Nannochloropsis cell wall, effectively increased the extraction rate of Nannochloropsis polar lipids, to solve the problem that the existing technology cannot extract from Microchloropsis Technical Problems of Obtaining Higher Purity EPA Polar Lipid Directly from Paeurococcus

Method used

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  • Method for extracting nannochloropsis oculata EPA polar lipid under assistance of enzyme treatment
  • Method for extracting nannochloropsis oculata EPA polar lipid under assistance of enzyme treatment
  • Method for extracting nannochloropsis oculata EPA polar lipid under assistance of enzyme treatment

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Experimental program
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Effect test

Embodiment 1

[0056] Mix Nannochloropsis with citric acid-sodium citrate buffer solution with pH=5 according to the ratio of solid to liquid 1:5, and add laccase so that the enzyme activity of laccase in the system is 0.24U / ml. Enzymolysis was carried out for 3 hours to obtain a Nannochloropsis enzymolysis suspension; the Nannochloropsis suspension was centrifuged at a centrifugation speed of 4000r / min and a centrifugation time of 20 minutes, and the supernatant was removed after centrifugation. Take the lower layer of sediment and dry at 65°C for 16 hours to obtain Nannochloropsis enzymatically hydrolyzed broken-wall algae powder; use Nannochloropsis enzymolyzed broken-walled algae powder according to the ratio of solid to liquid 1:15 and the concentration of 90 % ethanol is mixed, placed in a reaction kettle and sealed with nitrogen gas, heated to 70°C, extracted for 70 minutes, and extracted 8 times to obtain the Nannochloropsis polar lipid extract; extract the Nannochloropsis polar lipid...

Embodiment 2

[0058] Mix Nannochloropsis with citric acid-sodium citrate buffer solution with pH=5 according to the ratio of solid to liquid 1:5, and add laccase-cellulase complex enzyme to make the enzymatic activity of laccase and cellulase in the system 0.1U / ml and 100U / ml, respectively, and enzymolysis at 50°C for 3 hours to obtain a suspension of Nannochloropsis enzymolysis; centrifuge the suspension of Nannochloropsis at 4000r / min, Centrifuge at a centrifugation time of 20 minutes, remove the supernatant after centrifugation, and remove the lower layer of sediment at a drying temperature of 65°C for a drying time of 16 hours to obtain Nannochloropsis enzymatically broken wall algae powder; enzymatically hydrolyze Nannochloropsis The broken algae powder is mixed with 90% ethanol according to the ratio of material to liquid 1:15, placed in the reaction kettle and sealed with nitrogen gas, heated to 70°C, extracted for 70 minutes, and extracted 8 times to obtain Nannochloropsis Polar lip...

Embodiment 3

[0060] Mix Nannochloropsis with citric acid-sodium citrate buffer solution with pH=5 according to the ratio of solid to liquid 1:5, and add laccase-cellulase complex enzyme to make the enzymatic activity of laccase and cellulase in the system 1U / ml and 500U / ml, respectively, and enzymolysis at 50°C for 3 hours to obtain a Nannochloropsis enzymolysis suspension; centrifuge the Nannochloropsis hydrolysis suspension at a centrifugal speed of 4000r / min Centrifuge under the condition of 20 minutes, remove the supernatant after centrifugation, take the lower layer and precipitate at a drying temperature of 65°C, and dry for 16 hours to obtain Nannochloropsis enzymatically broken wall algae powder; The wall algae powder is mixed with 90% ethanol according to the ratio of solid to liquid at 1:15, placed in the reaction kettle and sealed with nitrogen gas, heated to 70°C, extracted for 70 minutes, and extracted 8 times to obtain Nannochloropsis. Lipid extract; Centrifuge the Nannochlor...

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Abstract

The invention relates to the technical field of biology, in particular to a method for extracting nannochloropsis oculata EPA polar lipid with assistance of enzyme treatment. The method comprises the following steps: 1, treating nannochloropsis oculata by using laccase to obtain nannochloropsis oculata enzymolysis suspension; 2, centrifuging the nannochloropsis oculata enzymolysis suspension, removing supernatant liquid, taking lower precipitate, and performing drying to obtain nannochloropsis oculata enzymolysis wall-broken algae powder; 3, carrying out subcritical extraction on the wall-broken nannochloropsis oculata powder to obtain a nannochloropsis oculata polar lipid extracting solution; 4, centrifuging the nannochloropsis oculata polar lipid extracting solution, taking supernatant liquid, and removing the solvent through vacuum evaporation to obtain the nannochloropsis oculata EPA polar lipid. The preparation process of the nannochloropsis oculata EPA polar lipid is safe, the method is convenient, and the extraction rate of the EPA polar lipid is remarkably improved compared with the prior art.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a method for extracting EPA polar lipid from Nannochloropsis algae assisted by enzyme treatment. Background technique [0002] Eicosapentaenoic Acid (EPA), as an omega-3 series of polyunsaturated fatty acids (Polyunsaturated fatty acids PUFAs) that cannot be synthesized by the human body, can prevent Alzheimer's disease, prevent brain function decline, and enhance immunity , Inhibit the synthesis of endogenous cholesterol and triglyceride, increase the activity of lipoprotein lipase and other biological activities. At present, most EPA products are mainly extracted from deep-sea fish oil and Antarctic krill oil as raw materials. [0003] The cell wall of Nannochloropsis is mainly composed of cellulose, hemicellulose, and lignin. The cell wall has a three-layer structure, and its strength is several times that of plant cell walls. The complex and tough cell wall reduces the e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C11B1/02C11B1/10C11B3/00
CPCC11B1/025C11B1/10C11B3/001
Inventor 张立伟郑竟成田华何东平张四红雷芬芬赵康宇张美兰
Owner WUHAN POLYTECHNIC UNIVERSITY
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