CN106834137A - A kind of arachidonic microbial fermentation processes - Google Patents
A kind of arachidonic microbial fermentation processes Download PDFInfo
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- CN106834137A CN106834137A CN201611268294.1A CN201611268294A CN106834137A CN 106834137 A CN106834137 A CN 106834137A CN 201611268294 A CN201611268294 A CN 201611268294A CN 106834137 A CN106834137 A CN 106834137A
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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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- 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
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- 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
A kind of arachidonic microbial fermentation processes, it is characterised in that solid medium is made up of following raw material：Beef extract-peptone, dusty yeast, maltose, vitamin A, vitamin K, sodium chloride, octyl methoxycinnamate, wheat flour；Fermentation medium is made up of following raw material：Maltose, beef extract-peptone, dusty yeast, sodium nitrate, potassium chloride, 12 hydrated sulfuric acid hydrogen potassium, vitamin B12, vitamin K, sodium acid carbonate, water.The arachidonic acid parent species obtained by solid material culture, the growth vigor of bacterium cell is strong, synchronism is preferable, physiological status is stable；By the optimization to zymotechnique, shorten fermentation series, simplify zymotechnique, effectively reduce fermenting and producing cost, arachidonic acid stable yield has recovery rate higher.
Description
Technical field
The present invention relates to a kind of arachidonic microbial fermentation processes, belong to field of microbial fermentation.
Background technology
Arachidonic acid, is all-cis formula-Arachidonic Acid, belongs to unrighted acid, wherein containing
Four carbon-to-carbon double bonds, a carbon-oxygen double bond, is higher unsaturated fatty acid.It is distributed widely in the animal kingdom, certain is present on a small quantity
In the glyceride of individual kind, can also be found in glycerophosphatide class.Can be synthesized by linoleic acid in human body.Infer that it is prostaglandin life
One of starting material of thing synthesis.
Arachidonic acid (Arachidonicacid) is Arachidonic acid, it be a kind of multivalence not
Saturated fatty acid, also referred to as Arachidonic acid, abbreviation ARA or AA.
Arachidonic acid is a kind of important essential fatty acid, itself can not be synthesized in vivo, it is necessary to supplied by food
Give, be the important precursor of human prostate's element synthesis, be also content highest in human body, be distributed a kind of most wide how unsaturated
Aliphatic acid, is primarily present in organ muscle and blood tissues, and being combined into struetural lipid with phosphatide plays an important role.Peanut four
Olefin(e) acid regulation cardiac excitability, participate in neuroendocrine, promote cell division, suppress platelet aggregation, anti-inflammatory, anticancer,
Lipid peroxidation inhibition, promote brain development and protection eyesight etc. the aspect there is the bioactivity of uniqueness, be widely used in food,
The field such as cosmetics, feed and its additive, babies ' formula milk powder, bio-pharmaceuticals.Arachidonic acid is widely present in animal body
In, adrenal gland, liver and sardine, egg yolk of animal etc. are mainly derived from, but content is relatively low, generally below 0.2% (wt/
wt)。
Because the arachidonic acid in human body is absorbed mainly by from the external world, so strengthening to arachidonic research and development
Have a very important role.Current people mainly obtain arachidonic acid using microbe fermentation method, and the expansion of seed is trained
Foster is the committed step of fermenting and producing, and the quality of parent species quality plays vital effect to the quality of production and yield, but existing
The most of parent species for having production arachidonic acid oil in technology are all to use liquid fermentation and culture method, in the product of acquisition still
Exist in grease that arachidonic acid content is relatively low, the growth of microorganism cycle is long, culture process is not up to optimization or fermentation institute
The problems such as culture medium cost of use is high, limits arachidonic large-scale industrial production.
The content of the invention
It is an object of the invention to provide a kind of arachidonic microbial fermentation processes, the liquid of its abandoning tradition is sent out
Ferment cultural method, using solid material culture arachidonic acid parent species, the growth vigor that obtains bacterium cell is strong, synchronism preferably, it is raw
Reason is in stable condition, can be mushroomed out after culture transferring to fermentation tank, shortens period of delay, and can keep the product recovery rate of stabilization.
The present invention is achieved in that described a kind of arachidonic microbial fermentation processes, its solid medium
It is made up of following raw material：Beef extract-peptone, dusty yeast, maltose, vitamin A, vitamin K, sodium chloride, methoxy cinnamic acid
Monooctyl ester, wheat flour.
Specifically, being made up of following raw material and its parts by weight, contain in every 100g culture mediums：Beef extract-peptone 30-
50g, dusty yeast 5-10g, maltose 20-25g, vitamin A 0.5-1g, vitamin K 0.25-0.5g, sodium chloride 1.2g, methoxyl group
Cinnamic acid monooctyl ester 0.5-0.8g, wheat flour are supplied.Octyl methoxycinnamate can improve the production effect of culture medium in the medium
Rate, strengthens bacterial activity, prevents culture medium aging.
Fermentation medium is made up of following raw material：Maltose, beef extract-peptone, dusty yeast, sodium nitrate, potassium chloride, ten
Two hydrated sulfuric acid hydrogen potassium, vitamin B12, vitamin K, sodium acid carbonate, water.
Specifically, above-mentioned fermentation medium is made up of following raw material and its parts by weight, contain in every 100g culture mediums：Wheat
Bud sugar 10-15g, beef extract-peptone 30-50g, dusty yeast 10-15g, sodium nitrate 1-3g, potassium chloride 1-2g, 12 hydrated sulfuric acids
Hydrogen potassium 0.5-1g, vitamin B120.25-0.5g, vitamin K 0.1-0.2g, sodium acid carbonate 1-2g, water are supplied.
It is as described below present invention also offers a kind of specific steps of arachidonic microbial fermentation processes：
（1）Seed culture is expected in the common Mortierella fermentation of producing arachidonic acid admittedly
（ⅰ）Actication of culture：Activated using slant strains, test tube slant culture medium is PDA solid mediums, by producing arachidonic acid
Common Mortierella strain be connected to PDA inclined-planes, 24-26 DEG C is cultivated 2-3 days.
（ⅱ）Admittedly expect the preparation of seed culture medium：Above-mentioned solid medium dispensing is mixed in proportion, 80-100 DEG C of boiling
20-30min, dries to not conglomeration of scattering, and is dispensed into specification as in KShi bottles of 1000mL, 121 DEG C sterilize by 200g/ bottles
30min。
（ⅲ）Admittedly expect the culture of seed：Fresh slant strains are added into spore under the aseptic washings of 15-25mL, spore is made and is hanged
Liquid, pours into the solid material culture medium that sterilized, and jog makes its mixing of trying one's best, and is cultivated 2-3 days in 24-26 DEG C, respectively at 12h and 24h
Respectively shake up once, as the parent species of one grade fermemtation.
（2）Fermented and cultured
（ⅰ）The preparation of fermentation medium, it is 5.8-6.0,121 DEG C of sterilizings that above-mentioned fermentation medium each component is mixed into regulation pH
30min。
（ⅱ）Shake flask fermentation culture, i.e. sample fermented and cultured
With the above-mentioned spore for expecting the surface of the seed admittedly under the aseptic washings of 200-300ml, spore suspension is made, is 5- by percent by volume
10% inoculum concentration accesses Medium of shaking flask fermentation, and cultivation temperature is 26-30 DEG C, and shaking speed 150-200r/min is cultivated 3-5 days；
Wet thallus are collected after fermentation ends, the grease extracted after drying in dry mycelium carries out gas chromatographic detection, detects arachidonic acid
Recovery rate.
（ⅲ）Ferment tank culture, i.e., middle sample fermented and cultured：With above-mentioned solid material kind sublist under the aseptic washings of 200-300ml
The spore in face, is made spore suspension, is that 5-10% inoculum concentrations access one grade fermemtation tank by percent by volume, and fermentation temperature is 26-30
DEG C, cultivate 1.5-2.5 days；One grade fermemtation to the inoculum concentration percent by volume of second order fermentation is 10-20%, fermentation temperature early stage control
System promotes thalli growth at 26-30 DEG C, and temperature, to promote arachidonic acid oil synthesis, is adjusted downward to 18-20 DEG C, training by the later stage
The cycle of supporting is 3-4.5 days；The need for ensure thalli growth and Synthetic Oil, whole fermentation process is except auto-feeding is through 118 DEG C
The percentage by weight of sterilizing 25min is 40% glucose solution, and percentage by weight is that 28% ammoniacal liquor is used to control reduced sugar to exist
Preceding 40h is added auxiliary to be that 3-8g/L and pH is outside 5.8-6.0, to be additionally carried out 2 times and add auxiliary material manually after 6-8g/L, 40h
The material time is respectively fermentation 36h and 69h, and auxiliary material is respectively by glucose, soybean-cake flour, corn starch, olive oil, dipotassium hydrogen phosphate
And all kinds of trace element compositions, trace element can be vitamin B1, vitamin B2, vitamin B6, vitamin B12, nicotinic acid, cigarette
The combination of one or more in acid amide, calcium pantothenate, folic acid, biotin, chlorophyll；Wet thallus are collected after fermentation ends, is dried
The grease extracted afterwards in dry mycelium carries out gas chromatographic detection, detects arachidonic acid recovery rate.
The beneficial effects of the invention are as follows the arachidonic acid parent species obtained by solid material culture, the growth of bacterium cell is lived
Power is strong, synchronism is preferable, physiological status is stable；By the optimization to zymotechnique, shorten fermentation series, simplify zymotechnique,
Fermenting and producing cost is effectively reduced, arachidonic acid stable yield has recovery rate higher.
Specific embodiment
A kind of arachidonic microbial fermentation processes of the present invention, are iurther discussed by following examples.
Embodiment 1
A kind of arachidonic microbial fermentation processes, its solid medium is made up of following raw material and its parts by weight, often
Contain in 100g culture mediums：Beef extract-peptone 45g, dusty yeast 6g, maltose 24g, vitamin A 0.6g, vitamin K 0.27g,
Sodium chloride 1.2g, octyl methoxycinnamate 0.7g, wheat flour are supplied.
A kind of arachidonic microbial fermentation processes, its fermentation medium is by following raw material and its weight portion array
Into, per 100g culture mediums in contain：Maltose 11g, beef extract-peptone 36g, dusty yeast 14g, sodium nitrate 2g, potassium chloride 2g,
12 hydrated sulfuric acid hydrogen potassium 0.8g, vitamin B120.29g, vitamin K 0.15g, sodium acid carbonate 2g, water are supplied.
Embodiment 2
A kind of arachidonic microbial fermentation processes, its solid medium is made up of following raw material and its parts by weight, often
Contain in 100g culture mediums：Beef extract-peptone 42g, dusty yeast 8g, maltose 25g, vitamin A 0.5g, vitamin K 0.5g,
Sodium chloride 1.2g, octyl methoxycinnamate 0.8g, wheat flour are supplied.
A kind of arachidonic microbial fermentation processes, its fermentation medium is by following raw material and its weight portion array
Into, per 100g culture mediums in contain：Maltose 10g, beef extract-peptone 50g, dusty yeast 15g, sodium nitrate 3g, potassium chloride 2g,
12 hydrated sulfuric acid hydrogen potassium 1g, vitamin B120.5g, vitamin K 0.2g, sodium acid carbonate 2g, water are supplied.
Embodiment 3
A kind of arachidonic microbial fermentation processes, its solid medium is made up of following raw material and its parts by weight, often
Contain in 100g culture mediums：Beef extract-peptone 30g, dusty yeast 5g, maltose 25g, retinol1 g, vitamin K 0.5g, chlorine
Change sodium 1.2g, octyl methoxycinnamate 0.5g, wheat flour to supply.
A kind of arachidonic microbial fermentation processes, its fermentation medium is by following raw material and its weight portion array
Into, per 100g culture mediums in contain：Maltose 15g, beef extract-peptone 37g, dusty yeast 15g, sodium nitrate 2.5g, potassium chloride
1.5g, 12 hydrated sulfuric acid hydrogen potassium 0.8g, vitamin B120.26g, vitamin K 0.17g, sodium acid carbonate 1.5g, water are supplied.
A kind of specific steps of arachidonic microbial fermentation processes, it is as described below：
（1）Seed culture is expected in the common Mortierella fermentation of producing arachidonic acid admittedly
（ⅰ）Actication of culture：Activated using slant strains, test tube slant culture medium is PDA solid mediums, by producing arachidonic acid
Common Mortierella strain be connected to PDA inclined-planes, 24-26 DEG C is cultivated 2-3 days.
（ⅱ）Admittedly expect the preparation of seed culture medium：Above-mentioned solid medium dispensing is mixed in proportion, 80-100 DEG C of boiling
20-30min, dries to not conglomeration of scattering, and is dispensed into specification as in KShi bottles of 1000mL, 121 DEG C sterilize by 200g/ bottles
30min。
（ⅲ）Admittedly expect the culture of seed：Fresh slant strains are added into spore under the aseptic washings of 15-25mL, spore is made and is hanged
Liquid, pours into the solid material culture medium that sterilized, and jog makes its mixing of trying one's best, and is cultivated 2-3 days in 24-26 DEG C, respectively at 12h and 24h
Respectively shake up once, as the parent species of one grade fermemtation.
（2）Fermented and cultured
（ⅰ）The preparation of fermentation medium, it is 5.8-6.0,121 DEG C of sterilizings that above-mentioned fermentation medium each component is mixed into regulation pH
30min。
（ⅱ）Shake flask fermentation culture, i.e. sample fermented and cultured
With the above-mentioned spore for expecting the surface of the seed admittedly under the aseptic washings of 200-300ml, spore suspension is made, is 5- by percent by volume
10% inoculum concentration accesses Medium of shaking flask fermentation, and cultivation temperature is 26-30 DEG C, and shaking speed 150-200r/min is cultivated 3-5 days；
Wet thallus are collected after fermentation ends, the grease extracted after drying in dry mycelium carries out gas chromatographic detection, detects arachidonic acid
Recovery rate.
（ⅲ）Ferment tank culture, i.e., middle sample fermented and cultured：With above-mentioned solid material kind sublist under the aseptic washings of 200-300ml
The spore in face, is made spore suspension, is that 5-10% inoculum concentrations access one grade fermemtation tank by percent by volume, and fermentation temperature is 26-30
DEG C, cultivate 1.5-2.5 days；One grade fermemtation to the inoculum concentration percent by volume of second order fermentation is 10-20%, fermentation temperature early stage control
System promotes thalli growth at 26-30 DEG C, and temperature, to promote arachidonic acid oil synthesis, is adjusted downward to 18-20 DEG C, training by the later stage
The cycle of supporting is 3-4.5 days；The need for ensure thalli growth and Synthetic Oil, whole fermentation process is except auto-feeding is through 118 DEG C
The percentage by weight of sterilizing 25min is 40% glucose solution, and percentage by weight is that 28% ammoniacal liquor is used to control reduced sugar to exist
Preceding 40h is added auxiliary to be that 3-8g/L and pH is outside 5.8-6.0, to be additionally carried out 2 times and add auxiliary material manually after 6-8g/L, 40h
The material time is respectively fermentation 36h and 69h, and auxiliary material is respectively by glucose, soybean-cake flour, corn starch, olive oil, dipotassium hydrogen phosphate
And all kinds of trace element compositions, trace element can be vitamin B1, vitamin B2, vitamin B6, vitamin B12, nicotinic acid, cigarette
The combination of one or more in acid amide, calcium pantothenate, folic acid, biotin, chlorophyll；Wet thallus are collected after fermentation ends, is dried
The grease extracted afterwards in dry mycelium carries out gas chromatographic detection, detects arachidonic acid recovery rate.
Embodiment 4
A kind of arachidonic microbial fermentation processes, its solid medium is made up of following raw material and its parts by weight, often
Contain in 100g culture mediums：Beef extract-peptone 40g, dusty yeast 6g, maltose 22g, vitamin A 0.9g, vitamin K 0.28g,
Sodium chloride 1.2g, octyl methoxycinnamate 0.6g, wheat flour are supplied.
A kind of arachidonic microbial fermentation processes, its fermentation medium is by following raw material and its weight portion array
Into, per 100g culture mediums in contain：Maltose 12g, beef extract-peptone 47g, dusty yeast 14g, sodium nitrate 2g, potassium chloride 1g,
12 hydrated sulfuric acid hydrogen potassium 0.6g, vitamin B120.37g, vitamin K 0.19g, sodium acid carbonate 1.8g, water are supplied.
A kind of specific steps of arachidonic microbial fermentation processes, it is as described below：
（1）Seed culture is expected in the common Mortierella fermentation of producing arachidonic acid admittedly
（ⅰ）Actication of culture：Activated using slant strains, test tube slant culture medium is PDA solid mediums, by producing arachidonic acid
Common Mortierella strain be connected to PDA inclined-planes, 24-26 DEG C is cultivated 2-3 days.
（ⅱ）Admittedly expect the preparation of seed culture medium：Above-mentioned solid medium dispensing is mixed in proportion, 80-100 DEG C of boiling
20-30min, dries to not conglomeration of scattering, and is dispensed into specification as in KShi bottles of 1000mL, 121 DEG C sterilize by 200g/ bottles
30min。
（ⅲ）Admittedly expect the culture of seed：Fresh slant strains are added into spore under the aseptic washings of 15-25mL, spore is made and is hanged
Liquid, pours into the solid material culture medium that sterilized, and jog makes its mixing of trying one's best, and is cultivated 2-3 days in 24-26 DEG C, respectively at 12h and 24h
Respectively shake up once, as the parent species of one grade fermemtation.
（2）Fermented and cultured
（ⅰ）The preparation of fermentation medium, it is 5.8-6.0,121 DEG C of sterilizings that above-mentioned fermentation medium each component is mixed into regulation pH
30min。
（ⅱ）Shake flask fermentation culture, i.e. sample fermented and cultured
With the above-mentioned spore for expecting the surface of the seed admittedly under the aseptic washings of 200-300ml, spore suspension is made, is 5- by percent by volume
10% inoculum concentration accesses Medium of shaking flask fermentation, and cultivation temperature is 26-30 DEG C, and shaking speed 150-200r/min is cultivated 3-5 days；
Wet thallus are collected after fermentation ends, the grease extracted after drying in dry mycelium carries out gas chromatographic detection, detects arachidonic acid
Recovery rate.
（ⅲ）Ferment tank culture, i.e., middle sample fermented and cultured：With above-mentioned solid material kind sublist under the aseptic washings of 200-300ml
The spore in face, is made spore suspension, is that 5-10% inoculum concentrations access one grade fermemtation tank by percent by volume, and fermentation temperature is 26-30
DEG C, cultivate 1.5-2.5 days；One grade fermemtation to the inoculum concentration percent by volume of second order fermentation is 10-20%, fermentation temperature early stage control
System promotes thalli growth at 26-30 DEG C, and temperature, to promote arachidonic acid oil synthesis, is adjusted downward to 18-20 DEG C, training by the later stage
The cycle of supporting is 3-4.5 days；The need for ensure thalli growth and Synthetic Oil, whole fermentation process is except auto-feeding is through 118 DEG C
The percentage by weight of sterilizing 25min is 40% glucose solution, and percentage by weight is that 28% ammoniacal liquor is used to control reduced sugar to exist
Preceding 40h is added auxiliary to be that 3-8g/L and pH is outside 5.8-6.0, to be additionally carried out 2 times and add auxiliary material manually after 6-8g/L, 40h
The material time is respectively fermentation 36h and 69h, and auxiliary material is respectively by glucose, soybean-cake flour, corn starch, olive oil, dipotassium hydrogen phosphate
And all kinds of trace element compositions, trace element can be vitamin B1, vitamin B2, vitamin B6, vitamin B12, nicotinic acid, cigarette
The combination of one or more in acid amide, calcium pantothenate, folic acid, biotin, chlorophyll；Wet thallus are collected after fermentation ends, is dried
The grease extracted afterwards in dry mycelium carries out gas chromatographic detection, detects arachidonic acid recovery rate.
Described above not limitation of the present invention, the present invention is also not limited to the example above.The art it is common
In essential scope of the invention, change, remodeling, addition or the replacement made should also belong to protection of the invention to technical staff
Scope.
Claims (5)
1. a kind of arachidonic microbial fermentation processes, it is characterised in that solid medium is made up of following raw material：Beef
Cream peptone, dusty yeast, maltose, vitamin A, vitamin K, sodium chloride, octyl methoxycinnamate, wheat flour.
2. a kind of arachidonic microbial fermentation processes according to claim 1, it is characterised in that solid medium
It is made up of following raw material and its parts by weight, is contained in every 100g culture mediums：Beef extract-peptone 30-50g, dusty yeast 5-10g,
Maltose 20-25g, vitamin A 0.5-1g, vitamin K 0.25-0.5g, sodium chloride 1.2g, octyl methoxycinnamate 0.5-
0.8g, wheat flour are supplied.
3. a kind of arachidonic microbial fermentation processes according to claim 1, it is characterised in that fermentation medium
It is made up of following raw material：Maltose, beef extract-peptone, dusty yeast, sodium nitrate, potassium chloride, 12 hydrated sulfuric acid hydrogen potassium, dimension life
Plain B12, vitamin K, sodium acid carbonate, water.
4. a kind of arachidonic microbial fermentation processes according to claim 3, it is characterised in that fermentation medium
It is made up of following raw material and its parts by weight, is contained in every 100g culture mediums：Maltose 10-15g, beef extract-peptone 30-
50g, dusty yeast 10-15g, sodium nitrate 1-3g, potassium chloride 1-2g, 12 hydrated sulfuric acid hydrogen potassium 0.5-1g, vitamin B120.25-
0.5g, vitamin K 0.1-0.2g, sodium acid carbonate 1-2g, water are supplied.
5. a kind of arachidonic microbial fermentation processes according to claim 1, it is characterised in that specific steps, such as
It is lower described：
1）Seed culture is expected in the common Mortierella fermentation of producing arachidonic acid admittedly
ⅰ）Actication of culture：Activated using slant strains, test tube slant culture medium is PDA solid mediums, by producing arachidonic acid
Common Mortierella strain be connected to PDA inclined-planes, 24-26 DEG C is cultivated 2-3 days.
ⅱ）Admittedly expect the preparation of seed culture medium：Above-mentioned solid medium dispensing is mixed in proportion, 80-100 DEG C of boiling 20-
30min, dries to not conglomeration of scattering, and is dispensed into KShi bottles that specification is 1000mL by 200g/ bottles, 121 DEG C of sterilizing 30min.
ⅲ）Admittedly expect the culture of seed：Fresh slant strains are added into spore under the aseptic washings of 15-25mL, spore suspension is made, poured into
Sterilized solid material culture medium, and jog makes its mixing of trying one's best, and is cultivated 2-3 days in 24-26 DEG C, and one is respectively shaken up respectively at 12h and 24h
It is secondary, as the parent species of one grade fermemtation.
2）Fermented and cultured
ⅰ）The preparation of fermentation medium, it is 5.8-6.0,121 DEG C of sterilizings that above-mentioned fermentation medium each component is mixed into regulation pH
30min。
ⅱ）Shake flask fermentation culture, i.e. sample fermented and cultured
With the above-mentioned spore for expecting the surface of the seed admittedly under the aseptic washings of 200-300ml, spore suspension is made, is 5- by percent by volume
10% inoculum concentration accesses Medium of shaking flask fermentation, and cultivation temperature is 26-30 DEG C, and shaking speed 150-200r/min is cultivated 3-5 days；
Wet thallus are collected after fermentation ends, the grease extracted after drying in dry mycelium carries out gas chromatographic detection, detects arachidonic acid
Recovery rate.
ⅲ）Ferment tank culture, i.e., middle sample fermented and cultured：With the above-mentioned the surface of the seed of material admittedly under the aseptic washings of 200-300ml
Spore, is made spore suspension, is that 5-10% inoculum concentrations access one grade fermemtation tank by percent by volume, and fermentation temperature is 26-30 DEG C,
Culture 1.5-2.5 days；One grade fermemtation to the inoculum concentration percent by volume of second order fermentation is 10-20%, and fermentation temperature beforehand control exists
26-30 DEG C, promote thalli growth, temperature, to promote arachidonic acid oil synthesis, is adjusted downward to 18-20 DEG C, culture week by the later stage
Phase is 3-4.5 days；The need for ensure thalli growth and Synthetic Oil, whole fermentation process is except auto-feeding is through 118 DEG C of sterilizings
The percentage by weight of 25min is 40% glucose solution, and percentage by weight is that 28% ammoniacal liquor is used to control reduced sugar in preceding 40h
For after 6-8g/L, 40h for 3-8g/L and pH be 5.8-6.0 outside, be additionally carried out 2 times and add auxiliary material manually, when adding auxiliary material
Between be respectively fermentation 36h and 69h, auxiliary material is respectively by glucose, soybean-cake flour, corn starch, olive oil, dipotassium hydrogen phosphate and each
Class trace element composition, trace element can be vitamin B1, vitamin B2, vitamin B6, vitamin B12, nicotinic acid, nicotinic acid
The combination of one or more in amine, calcium pantothenate, folic acid, biotin, chlorophyll；Wet thallus are collected after fermentation ends, after drying
Extracting the grease in dry mycelium carries out gas chromatographic detection, detects arachidonic acid recovery rate.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102373244A (en) * | 2011-11-30 | 2012-03-14 | 厦门金达威集团股份有限公司 | Microorganism fermentation method for arachidonic acid |
| CN104531787A (en) * | 2013-09-13 | 2015-04-22 | 厦门大学 | Method for preparing arachidonic acid (ARA) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102373244A (en) * | 2011-11-30 | 2012-03-14 | 厦门金达威集团股份有限公司 | Microorganism fermentation method for arachidonic acid |
| CN104531787A (en) * | 2013-09-13 | 2015-04-22 | 厦门大学 | Method for preparing arachidonic acid (ARA) |
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