CN110438173A - A kind of preparation method of arachidonic acid emulsion - Google Patents
A kind of preparation method of arachidonic acid emulsion Download PDFInfo
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- CN110438173A CN110438173A CN201910774878.3A CN201910774878A CN110438173A CN 110438173 A CN110438173 A CN 110438173A CN 201910774878 A CN201910774878 A CN 201910774878A CN 110438173 A CN110438173 A CN 110438173A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
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- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
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Abstract
The present invention provides a kind of preparation method of arachidonic acid emulsion, include the following steps: that (1) Mortierella alpina ferments；(2) fermentation liquid is subjected to filters pressing by sheet frame and obtains mycelia, then mycelia is cleaned, the filter cake for later forming mycelia crushes；(3) after the completion of crushing plus water reuses compound biological enzyme and carries out enzymolysis processing, maintains high-speed stirred in enzymolysis process, prevents conglomerate；(4) after the completion of enzymolysis processing, high speed shear is carried out to enzymolysis liquid using high speed shear head, up to arachidonic acid emulsion after homogeneous.Preparation method of the invention is directly obtained rich in arachidonic emulsion, it can be used directly as feedstuff or feed addictive after the emulsion is spray-dried, or emulsion directly extracts ARA grease after particular procedure, the risk that ARA is lost by high-temperature oxydation in smashed mycelia drying course in former traditional handicraft is reduced, and then ensure that the content and quality of ARA.
Description
Technical field
The present invention relates to arachidonic acid preparation technical fields, more particularly, to a kind of preparation of arachidonic acid emulsion
Method.
Background technique
Arachidonic acid (ARA), is all-cis formula-Arachidonic Acid, belongs to unsaturated fatty acid, wherein
Containing there are four carbon-to-carbon double bond, a carbon-oxygen double bond is higher unsaturated fatty acid.It is distributed widely in the animal kingdom, exists on a small quantity
In the glyceride of some kind, it can also find in glycerophosphatide class, can be synthesized by linoleic acid in human body.ARA is as breast milk
Component part, in neuro-physiology and neurology, the growth to baby be it is necessary, due to arachidonic in infants
The combined coefficient of acid is very low, and synthetic quantity is insufficient for the demand of body development, so must additionally supplement from food.
Substance rich in ARA is also used for feedstuff or feed addictive uses, but for the how unsaturateds rouge such as ARA
For fat acid, it can be again confronted with the risk of high-temperature oxydation in secondary processing process, obtain the content of ARA therein and quality not
To guarantee.
Summary of the invention
The object of the present invention is to provide a kind of preparation methods of arachidonic acid emulsion, solve more insatiable hungers such as existing ARA
The risk of high-temperature oxydation can be again confronted with during being processed with fatty acid, reduces the content of ARA therein and quality
Technical problem.
In order to achieve the above object, the invention provides the following technical scheme:
A kind of preparation method of arachidonic acid emulsion, includes the following steps:
(1) Mortierella alpina ferments；
(2) fermentation liquid is subjected to filters pressing by sheet frame and obtains mycelia, then mycelia is cleaned, later forms mycelia
Filter cake crush；
(3) after the completion of crushing plus water reuses compound biological enzyme and carries out enzymolysis processing, maintains high-speed stirred in enzymolysis process,
Prevent conglomerate；
(4) after the completion of enzymolysis processing, high speed shear is carried out to enzymolysis liquid using high speed shear head, it is high-pressure homogeneous rear up to institute
State arachidonic acid emulsion.
Further, in step (1), the preparation method of fermentation medium includes the following steps:
Prepare enriched medium:
In the distilled water for being 7.0 to pH value, 20~40mg/L of glucose, yeast powder 10~20mg/L, FeCl is added23~
20mg/L, Na2EDTA 1~10mg/L, H3BO31~10mg/L, MnCl2·4H2O 0.5~5mg/L, ZnSO4·7H2O 0.1
~0.6mg/L, CuSO4·5H2O 0.02~0.2mg/L, Na2MoO4·2H2O 0.01~0.8mg/L, CoCl2·6H2O
0.02~4mg/L obtains the enriched medium after the solution is heated to 40~60 DEG C of degree；
Prepare No. 1 growth medium
It takes 0.8~1.2mg/L of sodium glutamate, 0.8~1.2mg/L of propionic acid to be stirred, which is heated to 25~40
DEG C degree after obtain No. 1 growth medium；
Prepare No. 2 growth mediums
Take 9.5~10.5mg/L of sodium nitrate, 4~6mg/L of dipotassium hydrogen phosphate, 1.5~2.5mg/L of magnesium chloride, sodium chloride 19
~21mg/L, 0.8~1.2mg/L of yeast extract, is stirred, and obtains No. 2 lifes after which is heated to 40~60 DEG C of degree
Long culture medium；
By enriched medium obtained above, No. 1 growth medium and No. 2 growth mediums according to 1:0.3-0.5:0.2
Ratio pour into pH value be 7.0 clear water in stir evenly, after sterilizing be added Mortierella strain, stir evenly, in 20~30 DEG C
Degree, culture 3~4 days up to the fermentation medium.
Further, the sterilization method of fermentation medium is selected from high through 0.2 μm of aperture membrane filtration sterilizing or high temperature
One or both of pressure sterilizing.
Further, in step (3), by weight, the compound biological enzyme include 60 parts of biological enzyme, 22 parts of chitosan,
15 parts of sodium metasilicate, 6 parts of octyl glucoside, 4 parts of emulsifier, 2 parts of water retention agent, 6 parts of zinc gluconate.
Further, the biological enzyme includes dextranase, cellulase, amylase, beta-glucosidase, protease, paddy
Glutamine turns one of ammonia or a variety of.
The preparation method of arachidonic acid emulsion of the invention prepares arachidonic acid using Mortierella alpina mycelia,
It after everfermentation and enzymatic hydrolysis, directly obtains after carrying out high pressure homogenization rich in arachidonic emulsion, the emulsion is through spraying
It can be used directly as feedstuff or feed addictive after drying or emulsion directly extracts ARA after particular procedure
Grease reduces the risk that ARA is lost by high-temperature oxydation in smashed mycelia drying course in former traditional handicraft, and then guarantees
The content and quality of ARA.
Specific embodiment
Technical solution of the present invention is clearly and completely described below, it is clear that described embodiment is only this
Invention a part of the embodiment, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art exist
Every other embodiment obtained under the premise of creative work is not made, shall fall within the protection scope of the present invention.
Embodiment 1
In a kind of preferable embodiment of the present invention, the preparation method of the arachidonic acid emulsion includes the following steps:
(1) Mortierella alpina ferments；
The preparation method of fermentation medium includes the following steps:
Prepare enriched medium:
In the distilled water for being 7.0 to pH value, glucose 25mg/L, yeast powder 10mg/L, FeCl is added215mg/L,
Na2EDTA 3mg/L, H3BO32mg/L, MnCl2·4H2O 1mg/L, ZnSO4·7H2O 0.2mg/L, CuSO4·5H2O
0.06mg/L, Na2MoO4·2H2O 0.1mg/L, CoCl2·6H2O 0.1mg/L obtains institute after the solution is heated to 60 DEG C of degree
State enriched medium；
Prepare No. 1 growth medium
It takes sodium glutamate 0.8mg/L, propionic acid 0.8mg/L to be stirred, which is heated to obtaining after 40 DEG C of degree described
No. 1 growth medium；
Prepare No. 2 growth mediums
Take sodium nitrate 9.5mg/L, dipotassium hydrogen phosphate 4mg/L, magnesium chloride 1.5mg/L, sodium chloride 19mg/L, yeast extract
0.8mg/L is stirred, and obtains No. 2 growth mediums after which is heated to 40 DEG C of degree；
By enriched medium obtained above, No. 1 growth medium and No. 2 growth mediums according to the ratio of 1:0.3:0.2
Example is poured into the clear water that pH value is 7.0 and is stirred evenly, and Mortierella strain is added after sterilizing, stirs evenly, and is spent in 30 DEG C, culture 3
It is up to the fermentation medium.The sterilization method of fermentation medium is selected from and is sterilized through 0.2 μm of aperture membrane filtration.
(2) fermentation liquid is subjected to filters pressing by sheet frame and obtains mycelia, then mycelia is cleaned, later forms mycelia
Filter cake crush；
(3) after the completion of crushing plus water reuses compound biological enzyme and carries out enzymolysis processing, maintains high-speed stirred in enzymolysis process,
Prevent conglomerate；
By weight, the compound biological enzyme includes 60 parts of biological enzyme, 22 parts of chitosan, 15 parts of sodium metasilicate, octyl glucose
6 parts of glycosides, 4 parts of emulsifier, 2 parts of water retention agent, 6 parts of zinc gluconate.The biological enzyme includes dextranase, amylase, β-Portugal
Polyglycoside enzyme and protease.
(4) after the completion of enzymolysis processing, high speed shear is carried out to enzymolysis liquid using high speed shear head, up to the flower after homogeneous
Raw tetraenoic acid emulsion.
Embodiment 2
The fermentation medium preparation method that Mortierella alpina ferments in the present embodiment includes the following steps:
Prepare enriched medium:
In the distilled water for being 7.0 to pH value, glucose 30mg/L, yeast powder 10mg/L, FeCl is added25mg/L,
Na2EDTA 2mg/L, H3BO31mg/L, MnCl2·4H2O 2mg/L, ZnSO4·7H2O 0.5mg/L, CuSO4·5H2O
0.15mg/L, Na2MoO4·2H2O 0.2mg/L, CoCl2·6H2O 0.2mg/L obtains institute after the solution is heated to 50 DEG C of degree
State enriched medium；
Prepare No. 1 growth medium
It takes sodium glutamate 1.0mg/L, propionic acid 0.8mg/L to be stirred, which is heated to obtaining after 40 DEG C of degree described
No. 1 growth medium；
Prepare No. 2 growth mediums
Take sodium nitrate 10mg/L, dipotassium hydrogen phosphate 6mg/L, magnesium chloride 2mg/L, sodium chloride 20mg/L, yeast extract 1.1mg/
L is stirred, and obtains No. 2 growth mediums after which is heated to 60 DEG C of degree；
By enriched medium obtained above, No. 1 growth medium and No. 2 growth mediums according to the ratio of 1:0.4:0.2
Example is poured into the clear water that pH value is 7.0 and is stirred evenly, and Mortierella strain is added after sterilizing, stirs evenly, and is spent in 30 DEG C, culture 3
It is up to the fermentation medium.The sterilization method of fermentation medium is selected from through 0.2 μm of aperture membrane filtration sterilizing and height
Warm high pressure sterilization.
In step (3), by weight, compound biological enzyme includes 60 parts of biological enzyme, 22 parts of chitosan, 15 parts of sodium metasilicate, pungent
6 parts of base glucoside, 4 parts of emulsifier, 2 parts of water retention agent, 6 parts of zinc gluconate.The biological enzyme includes dextranase, cellulose
Enzyme and amylase.
Other steps are the same as embodiment 1.
Embodiment 3
The fermentation medium preparation method that Mortierella alpina ferments in the present embodiment includes the following steps:
Prepare enriched medium:
In the distilled water for being 7.0 to pH value, glucose 35mg/L, yeast powder 15mg/L, FeCl is added23mg/L,
Na2EDTA 6mg/L, H3BO33mg/L, MnCl2·4H2O 2mg/L, ZnSO4·7H2O 0.4mg/L, CuSO4·5H2O
0.1mg/L, Na2MoO4·2H2O 0.3mg/L, CoCl2·6H2O 0.3mg/L obtains institute after the solution is heated to 60 DEG C of degree
State enriched medium；
Prepare No. 1 growth medium
It takes sodium glutamate 1.2mg/L, propionic acid 0.8mg/L to be stirred, which is heated to obtaining after 30 DEG C of degree described
No. 1 growth medium；
Prepare No. 2 growth mediums
Take sodium nitrate 10.5mg/L, dipotassium hydrogen phosphate 5mg/L, magnesium chloride 2.5mg/L, sodium chloride 21mg/L, yeast extract
1.2mg/L is stirred, and obtains No. 2 growth mediums after which is heated to 60 DEG C of degree；
By enriched medium obtained above, No. 1 growth medium and No. 2 growth mediums according to the ratio of 1:0.5:0.2
Example is poured into the clear water that pH value is 7.0 and is stirred evenly, and Mortierella strain is added after sterilizing, stirs evenly, and is spent in 30 DEG C, culture 4
It is up to the fermentation medium.The sterilization method of fermentation medium is selected from through 0.2 μm of aperture membrane filtration sterilizing and height
Warm high pressure sterilization.
In step (3), by weight, compound biological enzyme includes 60 parts of biological enzyme, 22 parts of chitosan, 15 parts of sodium metasilicate, pungent
6 parts of base glucoside, 4 parts of emulsifier, 2 parts of water retention agent, 6 parts of zinc gluconate.The biological enzyme includes dextranase and albumen
Enzyme.
Other steps are the same as embodiment 1.
Embodiment 4
The fermentation medium preparation method that Mortierella alpina ferments in the present embodiment includes the following steps:
Prepare enriched medium:
In the distilled water for being 7.0 to pH value, glucose 40mg/L, yeast powder 10mg/L, FeCl is added23mg/L,
Na2EDTA 8mg/L, H3BO31mg/L, MnCl2·4H2O 1mg/L, ZnSO4·7H2O 0.6mg/L, CuSO4·5H2O
0.2mg/L, Na2MoO4·2H2O 0.3mg/L, CoCl2·6H2The solution is heated to obtaining after 40 DEG C of degree described by O 1mg/L
Enriched medium；
Prepare No. 1 growth medium
It takes sodium glutamate 0.8mg/L, propionic acid 1.2mg/L to be stirred, which is heated to obtaining after 40 DEG C of degree described
No. 1 growth medium；
Prepare No. 2 growth mediums
Take sodium nitrate 9.5mg/L, dipotassium hydrogen phosphate 6mg/L, magnesium chloride 2mg/L, 19~21g/L of sodium chloride, yeast extract
1.2mg/L is stirred, and obtains No. 2 growth mediums after which is heated to 60 DEG C of degree；
By enriched medium obtained above, No. 1 growth medium and No. 2 growth mediums according to the ratio of 1:0.4:0.2
Example is poured into the clear water that pH value is 7.0 and is stirred evenly, and Mortierella strain is added after sterilizing, stirs evenly, and is spent in 30 DEG C, culture 4
It is up to the fermentation medium.The sterilization method of fermentation medium is selected from through 0.2 μm of aperture membrane filtration sterilizing and height
Warm high pressure sterilization.
In step (3), by weight, compound biological enzyme includes 60 parts of biological enzyme, 22 parts of chitosan, 15 parts of sodium metasilicate, pungent
6 parts of base glucoside, 4 parts of emulsifier, 2 parts of water retention agent, 6 parts of zinc gluconate.The biological enzyme includes dextranase, protease
Turn ammonia with glutamine.
Other steps are the same as embodiment 1.
Embodiment 5
The fermentation medium preparation method that Mortierella alpina ferments in the present embodiment includes the following steps:
Prepare enriched medium:
In the distilled water for being 7.0 to pH value, glucose 20mg/L, yeast powder 20mg/L, FeCl is added215mg/L,
Na2EDTA 3mg/L, H3BO33mg/L, MnCl2·4H2O 3mg/L, ZnSO4·7H2O 0.2mg/L, CuSO4·5H2O
0.1mg/L, Na2MoO4·2H2O 0.2mg/L, CoCl2·6H2O 0.2mg/L obtains institute after the solution is heated to 60 DEG C of degree
State enriched medium；
Prepare No. 1 growth medium
It takes sodium glutamate 1.0mg/L, propionic acid 1.2mg/L to be stirred, which is heated to obtaining after 40 DEG C of degree described
No. 1 growth medium；
Prepare No. 2 growth mediums
Take sodium nitrate 10.5mg/L, dipotassium hydrogen phosphate 4mg/L, magnesium chloride 2mg/L, sodium chloride 21mg/L, yeast extract
1.2mg/L is stirred, and obtains No. 2 growth mediums after which is heated to 60 DEG C of degree；
By enriched medium obtained above, No. 1 growth medium and No. 2 growth mediums according to the ratio of 1:0.5:0.2
Example is poured into the clear water that pH value is 7.0 and is stirred evenly, and Mortierella strain is added after sterilizing, stirs evenly, and is spent in 30 DEG C, culture 4
It is up to the fermentation medium.The sterilization method of fermentation medium is selected from through 0.2 μm of aperture membrane filtration sterilizing and height
Warm high pressure sterilization.
In step (3), by weight, compound biological enzyme includes 60 parts of biological enzyme, 22 parts of chitosan, 15 parts of sodium metasilicate, pungent
6 parts of base glucoside, 4 parts of emulsifier, 2 parts of water retention agent, 6 parts of zinc gluconate.The biological enzyme includes dextranase, β-grape
Glycosidase, protease and glutamine turn ammonia.
Other steps are the same as embodiment 1.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (5)
1. a kind of preparation method of arachidonic acid emulsion, characterized by the following steps:
(1) Mortierella alpina ferments；
(2) fermentation liquid is subjected to filters pressing by sheet frame and obtains mycelia, then mycelia is cleaned, the filter for later forming mycelia
Cake crushes；
(3) after the completion of crushing plus water reuses compound biological enzyme and carries out enzymolysis processing, maintains high-speed stirred in enzymolysis process, prevents
Conglomerate；
(4) after the completion of enzymolysis processing, high speed shear is carried out to enzymolysis liquid using high speed shear head, it is high-pressure homogeneous rear up to the flower
Raw tetraenoic acid emulsion.
2. preparation method according to claim 1, it is characterised in that: in step (1), the preparation method packet of fermentation medium
Include following steps:
Prepare enriched medium:
In the distilled water for being 7.0 to pH value, 20~40mg/L of glucose, yeast powder 10~20mg/L, FeCl is added23~20mg/
L, Na2EDTA 1~10mg/L, H3BO31~10mg/L, MnCl2·4H2O 0.5~5mg/L, ZnSO4·7H2O 0.1~
0.6mg/L, CuSO4·5H2O 0.02~0.2mg/L, Na2MoO4·2H2O0.01~0.8mg/L, CoCl2·6H2O 0.02~
4mg/L obtains the enriched medium after the solution is heated to 40~60 DEG C of degree；
Prepare No. 1 growth medium
It takes 0.8~1.2mg/L of sodium glutamate, 0.8~1.2mg/L of propionic acid to be stirred, which is heated to 25~40 DEG C of degree
After obtain No. 1 growth medium；
Prepare No. 2 growth mediums
Take 9.5~10.5mg/L of sodium nitrate, 4~6mg/L of dipotassium hydrogen phosphate, 1.5~2.5mg/L of magnesium chloride, sodium chloride 19~
21mg/L, 0.8~1.2mg/L of yeast extract, is stirred, which is heated to obtain No. 2 growths after 40~60 DEG C of degree
Culture medium；
By enriched medium obtained above, No. 1 growth medium and No. 2 growth mediums according to the ratio of 1:0.3-0.5:0.2
Example is poured into the clear water that pH value is 7.0 and is stirred evenly, and Mortierella strain is added after sterilizing, stirs evenly, and is spent in 20~30 DEG C, training
3~4 days are supported up to the fermentation medium.
3. preparation method according to claim 2, it is characterised in that: be selected from warp for the sterilization method of fermentation medium
One or both of 0.2 μm of aperture membrane filtration sterilizing or autoclave sterilization.
4. preparation method according to claim 1, it is characterised in that: in step (3), by weight, the compound bio
Enzyme include 60 parts of biological enzyme, 22 parts of chitosan, 15 parts of sodium metasilicate, 6 parts of octyl glucoside, 4 parts of emulsifier, 2 parts of water retention agent,
6 parts of zinc gluconate.
5. the preparation method according to claim 4, it is characterised in that: the biological enzyme includes dextranase, cellulase, shallow lake
Powder enzyme, beta-glucosidase, protease, glutamine turn one of ammonia or a variety of.
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| CN112608851A (en) * | 2020-12-30 | 2021-04-06 | 润科生物工程（福建）有限公司 | Method for extracting oil contained in mortierella alpina |
| WO2021104164A1 (en) * | 2019-11-26 | 2021-06-03 | 瞿瀚鹏 | Mortierella alpina and use thereof, and microbial oil rich in ara at position sn-2, preparation method therefor and use thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2021104164A1 (en) * | 2019-11-26 | 2021-06-03 | 瞿瀚鹏 | Mortierella alpina and use thereof, and microbial oil rich in ara at position sn-2, preparation method therefor and use thereof |
| US12071647B2 (en) | 2019-11-26 | 2024-08-27 | Hanpeng QU | Mortierellaalpina strain and use thereof, microbial oil containing ARA at SN-2 position and preparation and uses thereof |
| CN112608851A (en) * | 2020-12-30 | 2021-04-06 | 润科生物工程（福建）有限公司 | Method for extracting oil contained in mortierella alpina |
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Application publication date: 20191112 |