CN102925502A - Industry method for producing arachidonic acid grease by using mortierella alpine - Google Patents
Industry method for producing arachidonic acid grease by using mortierella alpine Download PDFInfo
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- CN102925502A CN102925502A CN2011102281542A CN201110228154A CN102925502A CN 102925502 A CN102925502 A CN 102925502A CN 2011102281542 A CN2011102281542 A CN 2011102281542A CN 201110228154 A CN201110228154 A CN 201110228154A CN 102925502 A CN102925502 A CN 102925502A
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Abstract
The present invention provides an industry method for producing arachidonic acid grease by using mortierella alpine. The method is characterized by comprising the following steps: activating an original strain; carrying out seed culture; carrying out seed enlarge culture; and carrying out fermentation culture, wherein the fermentation culture comprises an initial fermentation stage, the seed enlarge culture liquid is inoculated in a fermentation tank to carry out stirring culture, glucose and edible oil are supplemented according to the situation, the initial fermentation stage is completed when bacterial concentration of the fermentation broth achieves 40% (by volume) and the dried mycelium total oil achieves 40 wt%, a post-fermentation stage starts, the fermentation broth is cooled to a temperature of 23-27 DEG C, sterile ammonia water is concurrently added to adjust the pH value to 8-9, standing fermentation is performed while the glucose content is controlled, the post-fermentation stage is completed when the arachidonic acid content is higher than 45 wt%, and the obtained fermentation broth is subjected to a fermentation treatment to obtain the grease containing the arachidonic acid. The industry method has the following characteristics that: a fermentation formula is simplified, specific yield of the arachidonic acid can be 10 gARA/L fermentation broth through the specific process control, a process is simple and easy to control, and industrial production is achieved.
Description
Technical field
The present invention relates to the arachidonic method of suitability for industrialized production, be specifically related under specific processing condition, utilize Mortierella alpina to expand and join the commercial run of producing arachidonic acid oil.
Background technology
Arachidonic acid (Arachidonic Acid, be called for short ARA) is n-6 series polyunsaturated fatty acid, it as synthesized human prostaglandin(PG), blood coagulation uh the precursor substance of alkane and leukotriene, have widely biological activity.Arachidonic acid also has important trophism, and is essential to baby's neural and physiological growth as lacto's natural component, comprised that by a plurality of expert bodies world health organisation recommendations adds in the infant formula powder as nutritious supplementary.Arachidonic acid is mainly derived from animal livers, fish oil, bovine adrenal etc., but arachidonic mass percentage concentration is very low in the animal tissues, only is 0.2%, and the source is very limited.Utilize the Production by Microorganism Fermentation arachidonic acid as the arachidonic new way of a kind of production, have and be not subjected to the advantages such as starting material and weather restriction, with short production cycle, technique simple, cost is lower, product content is high, separation and Extraction is easy, caused domestic and international researchist's extensive concern.
YASUHISA etc. research is found C/N than the form that can affect mycelia, when the C/N ratio less than 20 the time, size and the width of mycelium pellet are constant; The C/N ratio was greater than 20 o'clock, and the size of mycelium pellet and width can become large along with the increase of C/N ratio; The C/N ratio was at 15～20 o'clock, and arachidonic output is the highest.Higashiyama etc. have studied the impact of inorganic salt on Mortierella alpina mycelia form and ARA output, and the result shows: in the substratum that contains soyflour and glucose, add KH
2PO
4, Na
2SO
4, CaCl
2And MgCl
2Situation under, through optimizing, ARA output can reach 9.8g/L.Because arachidonic acid is present in the thalline lipid, the lipoid composition is the most important condition that increases the arachidonic acid yield in the increase thalline.The researchs such as A.Singh find to add analysis for soybean powder and vegetables oil can significantly improve thalline weight, fat content and the arachidonic acid yield that M.Alpina ATCC 3222 ferments, and ARA output reaches as high as 11.1g/L.Zhou Pengpeng etc. find that oil from Mucor does not have impact substantially on growth and the grease yield of Mortierella alpina, but can improve the content of ARA in grease, thereby can improve ARA output.
Summary of the invention
The invention provides a kind of Mortierella alpina that utilizes and produce arachidonic commercial run, the method can make arachidonic unit output reach 10g ARA/L fermented liquid by specific technology controlling and process.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
Utilize Mortierella alpina to produce the commercial run of arachidonic acid oil, it is characterized in that, it may further comprise the steps:
(1) activation of original strain: with Mortierella alpine trichoderma strain slant culture, obtain the spore suspension of Mortierella alpina;
(2) seed culture: the spore suspension of Mortierella alpina is inoculated in jolting cultivation acquisition seed culture fluid in the shake-flask culture base;
(3) seed enlarged culturing: according to the size of final fermentor tank, with the seed culture fluid of step (2) enlarged culturing step by step, obtain the seed scale-up medium;
(4) fermentation culture comprises initial fermentation stage and secondary fermentation stage:
Initial fermentation stage:
The seed scale-up medium of step (3) is inoculated in carries out stir culture in the fermentor tank, the main raw material component of described fermention medium is glucose 4～6%, yeast powder 1～3%, municipal tap water by mass percentage, pH value 5～7, in this initial fermenting process, add glucose control glucose content at 3～3.5wt% by stream, and in the initial 16h of fermentation, adding edible oil according to the foam production, the add-on of edible oil by volume percentage ratio is counted 0.1～0.3% of fermentating liquid volume;
Reach 40% (volume ratio) when the fermented liquid bacterium is dense, when the total oil of dry mycelium reached 40wt%, initial fermenting process finished, and enters the secondary fermentation stage;
The secondary fermentation stage:
Above-mentioned fermented liquid is cooled to 25 ± 2 ℃, add simultaneously aseptic ammoniacal liquor, regulating the pH value is 8～9, standing for fermentation, add glucose control glucose content at 1.0～1.5wt% by stream during the fermentation, and after arachidonic acid content is higher than 40wt%, stop to add glucose; When arachidonic content was higher than 45wt% in the fermented liquid, the secondary fermentation stage finished;
(5) fermented liquid is obtained containing arachidonic grease through the fermentation aftertreatment.
Press such scheme, described step (1) is: with the Mortierella alpina inoculation to the PDA slant medium, 28 ± 1 ℃ of lower cultivations 8～10 days, choose the eugonic PDA culture medium flat plate of mycelia and spore, take off mycelia and spore on the substratum, be mixed with spore suspension with sterilized water.
Press such scheme, described step (2) is: spore suspension is inoculated in the shake-flask culture base, cultivate 48h 28 ± 1 ℃ of lower joltings, shaking table jolting rotating speed is 200 ± 20 rev/mins, the mass percent of main raw material component is glucose 4～6%, peptone 1～2%, yeast extract 2～4%, matrix distilled water in the substratum, and the pH value is 7 ± 0.2.
Press such scheme, the mass percent of main raw material component is glucose 5～10%, yeast powder 1～3%, edible oil 0.1～0.3%, municipal tap water in the seed culture medium of described step (3), nature pH value, and carry out next step enlarged culturing when reaching 20% (volume ratio) bacterium is dense.
Press such scheme, every grade of incubation time in the described step (3) is 24～32h, and culture temperature is 30 ± 2 ℃, and air flow quantity is 0.8～1.0m
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.15～0.2MPa; The stirring velocity of described stir culture is preferably 50～70 and turns/min.
Press such scheme, the initial leavening temperature in the described step (4) is controlled to be that 30 ± 2 ℃, pH value change naturally, air flow quantity is 0.8～1.0m
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.2～0.3MPa.
Press such scheme, the glucose that stream adds in the described step (4) is the aseptic glucose solution of 10～30wt%.
Press such scheme, the stirring velocity of initial fermentation stage is 50～70 to turn/min in the described step (4).
Press such scheme, the air flow quantity in secondary fermentation stage is 1.0～1.5m in the described step (4)
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.2～0.3MPa.
Press such scheme, described step (5) is successively through deactivation, the separation of bacterium liquid, dry, microbial oil extraction process with fermented liquid.
Beneficial effect of the present invention:
(1) the inventive method only uses yeast powder and glucose as the Primary Fermentation raw material, has simplified fermentating formula, can effectively control the proferment material;
(2) by modulation process, can improve fermentation productivity, make the ARA output of every liter of fermented liquid reach 10g/L.The processing step of concrete regulation and control has: fermentation step is divided into initial fermentation stage and secondary fermentation stage, and for the different different control technique of stage employing, adopt stir culture in seed enlarged culturing and initial fermentation stage, be beneficial to the accumulation of fungal cell progenitor cells number, add vegetables oil at initial fermentation stage simultaneously, can be used on the one hand the control of earlier fermentation bio foam, also can be used as on the other hand the synthetic precursor of microbial oil and the supply of carbon source; Stop to stir in the secondary fermentation stage, standing for fermentation is conducive to the accumulation of mould grease and ARA content; The ammoniacal liquor that the secondary fermentation stage uses namely is pH adjusting agent, also is nitrogen source supplementing agent; Content by glucose in the controlled fermentation liquid can promote the accumulation of grease and the ability that arachidonic acid transforms during the fermentation; Adopt tall can to compress into capable fermentation culture, be conducive to improve dissolved oxygen level, thereby improve fermentation level;
(3) simple process is easy to control, realizes suitability for industrialized production.
Embodiment
Embodiment 1:
The activation of 1 original strain: will use peace doubly to manage the Mortierella alpina inoculation of preservation to the PDA slant medium, 28 ± 1 ℃ of lower cultivations 8 days, choose the eugonic PDA culture medium flat plate of mycelia and spore, take off mycelia and spore on the substratum, make spore suspension with sterilized water;
2 seed culture: spore suspension is inoculated in the shake-flask culture base, cultivate 48h 28 ± 1 ℃ of lower joltings, shaking table jolting rotating speed is 200 ± 20 rev/mins, the mass percent of main raw material component is glucose 4%, peptone 1%, yeast extract 3%, matrix distilled water in the liquid nutrient medium, and the pH value is 7.2;
3 seed enlarged culturing: final fermentor tank volume is 50m
3, so the selected seed tank is respectively 10m
3, 1m
3, the shake-flask seed fermented liquid of 1.5L step (2) is inoculated into 1m
3Seed enlarge in the fermentor tank, the mass percent of main raw material component is respectively glucose 6%, yeast powder 1.3%, edible oil 0.15%, municipal tap water in the seed culture medium, natural pH value, air capacity is controlled at 0.8～1.0m as required
3/ m
3Fermented liquid/min, the tank internal pressure is controlled at 0.17MPa, cultivates 24h at 30 ± 2 ℃, and the bacterium of fermentation cylinder for fermentation liquid is dense to be 25% (volume ratio), and this seed fermentation liquid is inoculated into 10m
3Seed enlarge fermentor tank and carry out the enlarged culturing second time, the mass percent that this seed enlarges seed culture medium main raw material component in the fermentor tank is glucose 5%, yeast powder 2%, edible oil 0.2%, municipal tap water, nature pH value, air capacity is controlled at 0.8～1.0m as required
3/ m
3Fermented liquid/min, the tank internal pressure is controlled at 0.19MPa, and stirring velocity is controlled to be 55 and turns/min, and behind the fermentation culture 28h, the bacterium of fermentation cylinder for fermentation liquid is dense to be 30%;
(4) the seed scale-up medium second time with step (3) is inoculated into 50m
3Fermentor tank in initially ferment, the main raw material component of this fermentation cylinder for fermentation substratum is glucose 4.6%, yeast powder 1.5%, municipal tap water by mass percentage, regulating the pH value with sodium hydroxide solution is 6.23, batch volume 25m
3Leavening temperature is controlled at that 30 ± 2 ℃, pH value change naturally, air capacity need to be controlled at 0.8～1.0m
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.25MPa, stirring velocity 60 turns/min, the control glucose content is at 3～3.5wt% in this initial fermenting process, glucose content is lower than 3wt% in fermented liquid, begins stream and adds concentration and stop when glucose content reaches 3.5wt% at the aseptic glucose solution of 10～30wt%; And in the initial 16h of fermentation, adding edible oil according to the foam production, edible oil add-on by volume percentage ratio is counted 0.1～0.3wt% of fermentating liquid volume;
Initial zymotechnique control sees the following form 1:
Table 1
Annotate: when bacterium is dense when reaching 40% (volume ratio), the microorganism cells number in the fermented liquid has reached requirement, and next step then is the stage of cell transformation accumulation grease, and the dense variation of bacterium is not very large, then need not dense the proceeding of bacterium to be detected again.
When fermentation culture to dense 40% (volume ratio) that reach of bacterium, the total oil of dry mycelium reaches 40wt%, initial fermenting process finishes;
The secondary fermentation stage: use fermentor tank external circulation heat exchanging system, the fermented liquid temperature of above-mentioned initial fermentation is down to 25 ± 2 ℃ in 24h, the aseptic ammoniacal liquor that in this process, adds simultaneously 0.01～0.02% (weight percentage), regulate pH value to 8.25, and use fermentor tank inner coil pipe interchanger to keep the fermented liquid temperature at 25 ± 2 ℃, air capacity is controlled at 1.0～1.5m as requested
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.25MPa, continue fermentation, control during the fermentation glucose content at 1.0～1.5wt%, beginning stream when glucose content is lower than 1.0wt%, to add concentration be that the aseptic glucose solution of 10～30wt% stops when glucose content reaches 1.5wt%, stops to mend sugar after arachidonic content is higher than 40wt%:
The secondary fermentation technology controlling and process sees the following form 2:
Table 2
When arachidonic content during greater than 45wt%, the secondary fermentation process can finish;
5 fermentation aftertreatments:
Deactivation: after fermentation ends, by high-temperature short-time sterilization equipment, fermented liquid is carried out the high-temperature instantaneous deactivation, the deactivation temperature is 122 ± 2 ℃, the deactivation 18s that holds time;
Bacterium liquid separates: the temperature of regulating fermented liquid by plate-type heat exchanger is 48 ± 2 ℃, sets sheet frame pressure 12MPa, and extrusion time 120s by sheet frame separate fermentation liquid, obtains filter cake, and the filter cake that then separates by pulverizer pulverizing sheet frame is wet thallus;
Dry: use the fluidized drying tower that wet thallus is carried out drying, obtain dry mycelium, 70～80 ℃ of control inlet temperature, every crowd of dry wet thalline weight 600kg, time of drying 120 ± 20min;
The statistics production data is as follows:
Fermentating liquid volume 41m
3
Dry mycelium weight is 1854kg;
The dry mycelium moisture content is 6.5wt%;
The total oil-contg of dry mycelium is 51wt%;
Arachidonic acid content 48.5wt% in the total oil of dry mycelium;
Unit fermented liquid output: amount/fermentating liquid volume of ARA in the dry mycelium=10.35gARA/L fermented liquid
Microbial oil extracts:
Use the arachidonic grease that contains in the acetone extract dry cell of microorganism, extraction temperature is not higher than 60 ℃, and a coextraction 7 times uses respectively amounts of acetone to see Table 4, for:
Table 4
| Extraction times | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| Acetone consumption m 3 | 4 | 3 | 2 | 2 | 2 | 2 | 2 |
Embodiment 2:
The activation of 1 original strain: will use peace doubly to manage the Mortierella alpina inoculation of preservation to the PDA slant medium, 28 ± 1 ℃ of lower cultivations 9 days, choose the eugonic PDA culture medium flat plate of mycelia and spore, take off mycelia and spore on the substratum, make spore suspension with sterilized water;
2 seed culture: spore suspension is inoculated in the shake-flask culture base, cultivate 48h 28 ± 1 ℃ of lower joltings, shaking table jolting rotating speed is 200 ± 20 rev/mins, the mass percent of main raw material component is glucose 5%, peptone 1.5%, yeast extract 2%, matrix distilled water in the liquid nutrient medium, and the pH value is 7.0;
3 seed enlarged culturing: final fermentor tank volume is 200m
3, so seeding tank is respectively 50m
3, 5m
3, 500L, the seed that 1L shake-flask seed fermented liquid is inoculated into 500L enlarges in the fermentor tank, cultivates 24h under 30 ± 2 ℃ temperature, and the bacterium of fermentation cylinder for fermentation liquid is dense to be 20% (volume ratio), and this fermented liquid is inoculated into 5m
3Fermentor tank in carry out the enlarged culturing 28h second time after, the bacterium of fermentation cylinder for fermentation liquid is dense to be 24% (volume ratio), continues fermented liquid is inoculated into 50m
3Fermentor tank in carry out for the third time enlarged culturing 24h after, the bacterium of fermentation cylinder for fermentation liquid is dense to be 30% (volume ratio), air capacity all was controlled at 0.8～1.0m as required when three grades of seeding tanks spread cultivation
3/ m
3Fermented liquid/min, the tank internal pressure is controlled at respectively 0.15MPa, 0.17MPa and 0.19MPa, and stirring velocity is 57 to turn/min.When three grades of seeding tanks spread cultivation in the seed culture medium mass percent of main raw material component be respectively glucose 6%, 6% and 5%, yeast powder 1.5%, 1.3% and 2%, edible oil 0.1%, 0.15% and 0.2%, municipal tap water, natural pH value;
The 4 for the third time seed scale-up mediums with step (3) are inoculated into 500m
3Fermentor tank in initially ferment, the main raw material component of this fermentation cylinder for fermentation substratum is glucose 5%, yeast powder 2%, municipal tap water by mass percentage, regulating the pH value with sodium hydroxide solution is 6.35, batch volume 120m
3Fermentation jar temperature is controlled at 30 ± 2 ℃, pH value change naturally, air capacity need to be controlled at 0.8～1.0m
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.27MPa, stirring velocity 65 turns/min, the control glucose content is at 3～3.5wt% in this initial fermenting process, glucose content is lower than 3wt% in fermented liquid, begins stream and adds concentration and stop when glucose content reaches 3.5wt% at the aseptic glucose solution of 10～30wt%; And in fermentation 16h, adding edible oil according to the foam production, edible oil add-on by volume percentage ratio is counted 0.1～0.3wt% of fermentating liquid volume;
Initial zymotechnique control sees the following form 5:
Table 5
When fermentation culture to dense 40% (volume ratio) that reach of fermented liquid bacterium, the total oil of dry mycelium reaches 40wt%, initial fermenting process finishes;
The secondary fermentation stage: use fermentor tank external circulation heat exchanging system, the fermented liquid temperature is down to 25.3 ℃ in 24h, simultaneously in this process, add ammoniacal liquor, regulate pH value to 8.1, and use fermentor tank inner coil pipe interchanger to keep the fermented liquid temperature at 25 ± 2 ℃, air capacity is controlled at 1.0～1.5m as requested
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.3MPa, continue fermentation, control during the fermentation glucose content at 1.0～1.5wt%, when glucose content is lower than 1.0wt%, beginning stream, to add concentration be that the aseptic glucose solution of 10～30wt% stops when glucose content reaches 1.5wt%, stops to mend sugar after arachidonic content is higher than 40wt%:
The secondary fermentation technology controlling and process sees the following form 6:
Table 6
When arachidonic acid content reached 45wt% in the dry mycelium grease, the secondary fermentation process can finish;
5 fermentation aftertreatments:
Deactivation: after fermentation ends, by high-temperature short-time sterilization equipment, fermented liquid is carried out the high-temperature instantaneous deactivation, the deactivation temperature is 122 ± 2 ℃, the deactivation 15s that holds time;
Bacterium liquid separates: is adjusted to 48 ± 2 ℃ by plate-type heat exchanger, sets sheet frame pressure 10MPa, and extrusion time 120s, by sheet frame separate fermentation liquid, obtaining filter cake then is wet thallus by the filter cake that pulverizer pulverizing sheet frame separates.
Dry: use the fluidized drying tower that wet thallus is carried out drying, obtain dry mycelium, 70～80 ℃ of control inlet temperature, every crowd of dry wet thalline weight 600kg, time of drying 120 ± 20min;
The statistics production data is as follows:
Fermentating liquid volume 170m
3
Dry mycelium weight is 7430kg;
The moisture content of dry mycelium is 5.76wt%;
Total oil-contg 50.5wt% of dry mycelium;
Arachidonic content 49.3wt% in the total oil of dry mycelium;
Unit fermented liquid output: amount/fermentating liquid volume of ARA in the dry mycelium=10.25g ARA/L fermented liquid
Microbial oil extracts:
Use in the acetone extract dry cell of microorganism and contain arachidonic grease, extraction temperature is not higher than 60 ℃, and a coextraction 6 times uses respectively amounts of acetone to see Table 7, for:
Table 7
| Extraction times | 1 | 2 | 3 | 4 | 5 | 6 |
| Acetone consumption m 3 | 14 | 7 | 4 | 4 | 4 | 4 |
Claims (10)
1. utilize Mortierella alpina to produce the commercial run of arachidonic acid oil, it is characterized in that, it may further comprise the steps:
(1) activation of original strain: with Mortierella alpine trichoderma strain slant culture, obtain the spore suspension of Mortierella alpina;
(2) seed culture: the spore suspension of Mortierella alpina is inoculated in jolting cultivation acquisition seed culture fluid in the shake-flask culture base;
(3) seed enlarged culturing: according to the size of final fermentor tank, with the seed culture fluid of step (2) enlarged culturing step by step, obtain the seed scale-up medium;
(4) fermentation culture comprises initial fermentation stage and secondary fermentation stage:
Initial fermentation stage:
The seed scale-up medium of step (3) is inoculated in carries out stir culture in the fermentor tank, the chief component composition of described fermention medium is glucose 4 ~ 6%, yeast powder 1 ~ 3%, municipal tap water by mass percentage, pH value 5 ~ 7, in this initial fermenting process, add glucose control glucose content at 3 ~ 3.5wt% by stream, and in the initial 16h of fermentation, adding edible oil according to the foam production, the add-on of edible oil by volume percentage ratio is counted 0.1 ~ 0.3% of fermentating liquid volume;
Reach the 40%(volume ratio when the fermented liquid bacterium is dense), when the total oil of dry mycelium reached 40wt%, initial fermenting process finished, and enters the secondary fermentation stage;
The secondary fermentation stage:
Above-mentioned fermented liquid is cooled to 25 ± 2 ℃, add simultaneously aseptic ammoniacal liquor, regulating the pH value is 8 ~ 9, standing for fermentation, add glucose control glucose content at 1.0 ~ 1.5wt% by stream during the fermentation, and after arachidonic acid content is higher than 40wt%, stop to add glucose; When arachidonic content was higher than 45wt% in the fermented liquid, the secondary fermentation stage finished;
(5) fermented liquid is obtained containing arachidonic grease through the fermentation aftertreatment.
2. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1, it is characterized in that, described step (1) is: with the Mortierella alpina inoculation to the PDA slant medium, 28 ± 1 ℃ of lower cultivations 8 ~ 10 days, choose the eugonic PDA culture medium flat plate of mycelia and spore, take off mycelia and spore on the substratum, be mixed with spore suspension with sterilized water.
3. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1, it is characterized in that, described step (2) is: spore suspension is inoculated in the shake-flask culture base, cultivate 48h 28 ± 1 ℃ of lower joltings, shaking table jolting rotating speed is 200 ± 20 rev/mins, the mass percent of main raw material component is glucose 4 ~ 6%, peptone 1 ~ 2%, yeast extract 2 ~ 4%, matrix distilled water in the substratum, and the pH value is 7 ± 0.2.
4. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1, it is characterized in that, the mass percent of main raw material component is glucose 5 ~ 10%, yeast powder 1 ~ 3%, edible oil 0.1 ~ 0.3%, municipal tap water in the seed culture medium of described step (3), nature pH value, and in the dense 20%(of the reaching volume ratio of bacterium) carries out next step enlarged culturing the time.
5. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1 is characterized in that, every grade of incubation time in the described step (3) is 24 ~ 32h, and culture temperature is 30 ± 2 ℃, and air flow quantity is 0.8 ~ 1.0 m
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.15 ~ 0.2MPa.
6. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1 is characterized in that, the initial leavening temperature in the described step (4) is controlled to be that 30 ± 2 ℃, pH value change naturally, air flow quantity is 0.8 ~ 1.0m
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.2 ~ 0.3MPa.
7. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1 is characterized in that, the glucose that stream adds in the described step (4) is the aseptic glucose solution of 10 ~ 30wt%.
8. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1 is characterized in that, the stirring velocity of initial fermentation stage is 50 ~ 70 to turn/min in the described step (4).
9. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1 is characterized in that, the air flow quantity in secondary fermentation stage is 1.0 ~ 1.5m in the described step (4)
3/ m
3Fermented liquid/min, the tank internal pressure is at 0.2 ~ 0.3MPa.
10. the commercial run that utilizes Mortierella alpina to produce arachidonic acid oil according to claim 1 is characterized in that, described step (5) is successively through deactivation, the separation of bacterium liquid, dry, microbial oil extraction process with fermented liquid.
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| CN103451247A (en) * | 2013-09-13 | 2013-12-18 | 厦门大学 | Preparation method of arachidonic acid |
| CN103882071A (en) * | 2014-03-14 | 2014-06-25 | 嘉必优生物工程（武汉）有限公司 | Microbial oil and preparation method thereof |
| CN104651423A (en) * | 2015-01-28 | 2015-05-27 | 江南大学 | Method for fermentation production of arachidonic acid by adopting two-stage pH control |
| CN104805139A (en) * | 2015-03-20 | 2015-07-29 | 湖北产业技术创新与育成中心 | Method for increasing yield of ARA (arachidonic acid) produced through fermentation of Mortierella alpina by adding porphyridium |
| CN108410916A (en) * | 2018-05-30 | 2018-08-17 | 湖北福星生物科技有限公司 | The production method of arachidonic acid oil |
| CN109666710A (en) * | 2018-12-29 | 2019-04-23 | 嘉必优生物技术（武汉）股份有限公司 | Arachidonic acid oil product, selenium-rich Mortierella alpina and the preparation method of selenium-rich |
| CN110129384A (en) * | 2018-02-08 | 2019-08-16 | 浙江海正药业股份有限公司 | A kind of arachidonic preparation method |
| CN110438173A (en) * | 2019-08-21 | 2019-11-12 | 湖北福星生物科技有限公司 | A kind of preparation method of arachidonic acid emulsion |
| CN111575323A (en) * | 2017-02-23 | 2020-08-25 | 内蒙古金达威药业有限公司 | Method for extracting ARA crude oil |
| CN112625999A (en) * | 2020-12-30 | 2021-04-09 | 嘉必优生物技术（武汉）股份有限公司 | Method for domesticating filamentous fungi through continuous fermentation |
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| CN103451247B (en) * | 2013-09-13 | 2015-08-19 | 厦门大学 | A kind of arachidonic preparation method |
| CN103882071B (en) * | 2014-03-14 | 2016-10-05 | 嘉必优生物技术（武汉）股份有限公司 | A kind of microbial oil and preparation method thereof |
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| CN104805139A (en) * | 2015-03-20 | 2015-07-29 | 湖北产业技术创新与育成中心 | Method for increasing yield of ARA (arachidonic acid) produced through fermentation of Mortierella alpina by adding porphyridium |
| CN111575323A (en) * | 2017-02-23 | 2020-08-25 | 内蒙古金达威药业有限公司 | Method for extracting ARA crude oil |
| CN110129384A (en) * | 2018-02-08 | 2019-08-16 | 浙江海正药业股份有限公司 | A kind of arachidonic preparation method |
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| CN108410916A (en) * | 2018-05-30 | 2018-08-17 | 湖北福星生物科技有限公司 | The production method of arachidonic acid oil |
| CN109666710A (en) * | 2018-12-29 | 2019-04-23 | 嘉必优生物技术（武汉）股份有限公司 | Arachidonic acid oil product, selenium-rich Mortierella alpina and the preparation method of selenium-rich |
| CN110438173A (en) * | 2019-08-21 | 2019-11-12 | 湖北福星生物科技有限公司 | A kind of preparation method of arachidonic acid emulsion |
| CN112625999A (en) * | 2020-12-30 | 2021-04-09 | 嘉必优生物技术（武汉）股份有限公司 | Method for domesticating filamentous fungi through continuous fermentation |
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