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Technology for extracting arachidonic acid oil by aqueous enzymatic extraction

A technology of arachidonic acid and water enzymatic method, applied in the direction of producing fat, fat oil/fat production, etc., can solve the problems of difficult to realize industrialization, large amount of cooling water, difficult to extract in batches, etc., and achieves simple equipment, reduced operating costs, Applicable effect

Active Publication Date: 2010-10-27
NANJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, there are various shortcomings and deficiencies in these oil extraction methods: (1) the Soxhlet extraction method takes a long time, consumes a large amount of cooling water, and is difficult to extract in batches, and is only limited to laboratory-scale oil extraction
(2) The oil yield of the organic solvent method is low, and the oil quality is poor. For arachidonic acid, the residue of organic solvents has potential problems of food safety
(3) Ultrasonic extraction requires large power and small processing scale, making it difficult to realize industrialization
(4) Supercritical CO 2 Extraction, complex equipment, high production costs, limited to laboratory-scale oil extraction
Therefore, it is necessary to seek an oil extraction method with high oil yield, high oil quality, high efficacy, low energy consumption and easy industrialization. The extraction process of arachidonic acid oil from Mortierella alpina thalline using aqueous enzymatic method has not been found. Literature report

Method used

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  • Technology for extracting arachidonic acid oil by aqueous enzymatic extraction
  • Technology for extracting arachidonic acid oil by aqueous enzymatic extraction
  • Technology for extracting arachidonic acid oil by aqueous enzymatic extraction

Examples

Experimental program
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Embodiment 1

[0036] Embodiment 1: strain fermentation culture

[0037] The medium formula is as follows:

[0038] Seed medium: 100g / L glucose, 11g / L yeast powder, 8g / L corn steep liquor, 3.8g / L KH 2 PO 4 , the initial pH is 5.0-6.0.

[0039] Fermentation medium: 100g / L glucose, 11g / L yeast powder, 3.8g / L KH 2 PO 4 , 3.4g / L NaNO 3 , 0.5g / LMgSO 4 .7H 2 O. The initial pH is 5.5-6.5.

[0040] 1) Activation of strains

[0041] Before preparing the seeds, the PDA slant is used to activate the strains, and the activated strains are transferred to the PDA solid medium for 3-5 days to accumulate a large number of spores for future use.

[0042] 2) The seeds are enlarged once

[0043]After the spores of Mortierella alpina grown on the PDA solid medium were eluted with sterile water, they were inserted into a grooved shake flask filled with seed medium through aseptic operation. The liquid volume of the shake bottle is 50ml / 250ml. Seed culture conditions: the rotation speed of the shaker...

Embodiment 2

[0049] According to Example 1, ferment and cultivate the thallus, add distilled water to the fermentation liquid and dilute it by 2 volume multiples to make a thallus suspension, adjust the pH value to 6.0, divide it into 50mL test tubes according to 30mL equal amounts, add 20mg of helicase, and heat at 40°C Shake at constant temperature for 8 hours; add 4 mL of ethanol and 10 mL of n-hexane for the first extraction, centrifuge at 5000 rpm for 5 min to take the organic phase, add 10 mL of n-hexane to the remaining aqueous phase and solid phase for secondary extraction, and centrifuge at 5000 rpm for 5 min to get the organic phase , the combined organic phase was rotary evaporated to obtain 0.4854g of organic solvent and arachidonic acid oil, and the control group Soxhlet extraction method obtained 0.4994g of oil, and the oil yield was 97.2%.

Embodiment 3

[0051] According to Example 1, ferment and cultivate the thallus, add distilled water to the fermentation broth to dilute 2 volume multiples to make a thallus suspension, adjust the pH value to 6.0, divide into 50mL test tubes according to 30mL equal amounts, add 20mg of helicase, and heat at 37°C Shake at constant temperature for 8 hours; add 4.5mL of ethanol and 10mL of n-hexane for the first extraction, centrifuge at 5000rpm for 5min to take the organic phase, add 10mL of n-hexane to the remaining aqueous phase and solid phase for secondary extraction, centrifuge at 5000rpm for 5min to take the organic phase Phase, combined organic phase rotary evaporation to obtain organic solvent and arachidonic acid oil 0.4752g, control group Soxhlet extraction method to obtain oil 0.4994g, oil yield was 95.1%.

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Abstract

The invention discloses a technology for extracting arachidonic acid oil by an aqueous enzymatic extraction. The technology comprises the following steps: using Mortierella alpina as an original strain, fermenting and culturing, diluting by adding distilled water to obtain a thallus suspension, regulating the pH value of the suspension, adding a biological enzyme, oscillating at constant temperature for 6-10 hours, adding ethanol and normal hexane to carry out oil extraction, centrifugating to delaminate, taking out an organic solvent phase, adding normal hexane into the left water phase and solid phase to carry out secondary extraction, centrifugating to delaminate, combining with the organic solvent phase, and carrying out rotary evaporation on the organic solvent phase to obtain the oil and an organic solvent. The invention has the advantages of low energy consumption, low cost, high yield of arachidonic acid oil, high quality, simple reactor, easy batch large-scale application and the like.

Description

technical field [0001] The invention relates to a process for producing arachidonic acid (ARA) oil by using Mortierella alpina, and belongs to the field of bioengineering. Specifically, the method of using biological enzymes to break the cell wall of Mortierella alpina to extract arachidonic acid oil is a technical field of separation and extraction in the downstream process of bioengineering. Background technique [0002] Arachidonic acid (ARA, namely 5,8,11,14-eicosatetraenoic acid) belongs to the omega-6 series of long-chain polyunsaturated fatty acids (PUFAs), and is the most active and essential Polyunsaturated fatty acids have the functions of promoting intellectual development, improving human vision, lowering blood lipids, enhancing immunity and anti-cancer, etc., and are widely used in functional health care, biomedicine, cosmetics and feed additives. Since arachidonic acid oil is synthesized in a very small amount in adults and cannot be synthesized in infants and...

Claims

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Application Information

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IPC IPC(8): C11B1/00C11B1/10
Inventor 黄和尤江英彭超纪晓俊路金淼刘欣丛蕾蕾黎志勇
Owner NANJING UNIV OF TECH
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