CN104278107B - A kind of method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil - Google Patents
A kind of method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil Download PDFInfo
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- CN104278107B CN104278107B CN201410556081.3A CN201410556081A CN104278107B CN 104278107 B CN104278107 B CN 104278107B CN 201410556081 A CN201410556081 A CN 201410556081A CN 104278107 B CN104278107 B CN 104278107B
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- 230000004151 fermentation Effects 0.000 title claims abstract description 107
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- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 title claims abstract description 77
- 229910052760 oxygen Inorganic materials 0.000 title claims abstract description 77
- 239000001301 oxygen Substances 0.000 title claims abstract description 77
- 235000021342 arachidonic acid Nutrition 0.000 title claims abstract description 47
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- 238000013019 agitation Methods 0.000 description 5
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention relates to a kind of method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil.Mortierella alpine trichoderma strain is amplified cultivation step by step through test tube slant culture medium and Shake flask medium, prepares seed liquor, be then inoculated in fermentation tank and carry out fermented and cultured, whole sweat is divided into earlier fermentation, ferment middle and 3 stages of fermentation later stage；Rotating speed is initially set 0rpm, and dissolved oxygen is initially set in 100%, at earlier fermentation so that dissolved oxygen is stable 40% 70% after declining；At ferment middle, dissolved oxygen is controlled 20% 40%；Phase after fermentation, dissolved oxygen is controlled 40% 70%.Fermentation ends can make Mortierella alpina dry cell weight, fat content, arachidonic acid account for the percentage composition of total grease, arachidonic specific yield can reach respectively 55g/L, the 54%th, 65%, 20.628g/L, arachidonic acid produces intensity and reaches 2.292g/ (L*d).
Description
Technical field
The present invention relates to a kind of method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil, belong to micro-
Technical field of biological fermentation.
Background technology
Since the nineties in 20th century, health diet and nutrition arrangement are increasingly paid attention to by people, and how unsaturated long-chain is
Aliphatic acid progresses into the sight line of people.Long-chain polyunsaturated fatty acid mainly includes arachidonic acid (ARA), 22 carbon six
Olefin(e) acid (DHA) and eicosapentaenoic acid (EPA).These aliphatic acid have not only played important work in component film structure of phospholipid composition
With, and can be as the precursor of composite signal molecule eicosanoid (including prostaglandin, thromboxane and leukotriene etc.)
Material, is widely used in medicine and cosmetics, especially infant food and nutritional field.
At present, the technique of Production by Microorganism Fermentation arachidonic acid oil comes into the industrial production stage.But, send out
Ferment method produces and is also encountered by problems, and such as fermentation method cycle length, inversion rate of glucose is low, cellular biomass is low, grease contains
Measuring low, saturated and monounsaturated fatty acids too high levels, especially in sweat, thalli morphology is difficult to control to.
The fermentation tank using in traditional mode of production mainly includes mechanical agitation type and gas-lifting type etc..Mechanical agitation type fermentation tank makes
High with power, energy consumption is big, and blade shearing force is relatively big simultaneously, has a significant impact thalli morphology, it is easy to smash thalline, in broken
Featheriness.On the other hand, airlift fermentor enters filtrated air as motive force with drum, does not rolls zymotic fluid, not shearing
Power, but fermentation later stage Fungal biodiversity length is to a certain extent, and zymotic fluid density is big, and viscosity is high, and the promotion of gas is to zymotic fluid
The influence degree seething mixing is obviously reduced, and dissolved oxygen is wayward, and thalli morphology is wayward equally.But, thalli morphology pair
The indexs such as Fungal biodiversity, fat content and arachidonic acid percentage composition have directly impact.
The general level of current Mortierella alpina fermentation producing arachidonic acid oil is: 10-16 days cycle, biomass 22-
37g/L, arachidonic acid accounts for the 33-60% of total fat content.One group of the highest experiment of the arachidonic acid content wherein reported
The content that 12.1 days cycles, biomass 38.3g/L, oil content 26.4g/L, arachidonic acid account for total grease reaches 75%.But,
Arachidonic production intensity is substantially at 0.3-1.45g/ (L*d), and the peak of arachidonic acid net content report is also only
For 18g/L.In view of fermentation period length, easily cause microbiological contamination, production intensity to be still within the problems such as reduced levels, have and carry further
The space rising.Additionally, the at present higher experiment of the arachidonic acid net content of report be substantially 250mL shaking flask, 1L, 5L,
Carry out in the level of 20L small fermentor, seldom have industry to apply preferable example to report in amplifying.
Therefore, the drawback affecting thalli morphology in view of existing mechanical agitation type fermentation tank and airlift fermentor is right
In the fermentation that Mortierella alpina fermentation producing arachidonic acid oil such cycle length, fabulous oxygen and fermentation later stage viscosity significantly raise
System, optimizes suitable high cell density fermentation condition, provides out one can be suitable for Mortierella alpina thalline accumulation yield peanut
The novel process of tetraenoic acid grease, promotes Fungal biodiversity and arachidonic acid oil content, improves arachidonic acid and produces strong
Degree, and it is actually necessary to be committed to industrialized production.
Content of the invention
In view of the problems referred to above of prior art, the purpose of the present invention is to propose to one and send out based on Dissolved oxygen regulation Mortierella alpina
The method of ferment producing arachidonic acid oil, it is by with thalli morphology as target, in conjunction with mechanical agitation type fermentation tank and gas-lifting type
The advantage of fermentation tank, by the regulation and control to technique, improves fermentation productivity.
To achieve these goals, the technical solution used in the present invention is:
A kind of method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil, by Mortierella alpine trichoderma strain
Amplify cultivation through test tube slant and shaking flask step by step, prepare seed liquor, be then inoculated in fermentation tank and carry out fermented and cultured, sending out
During ferment, to control Mortierella alpina fermentation form as target, with dissolved oxygen parameter of fermenting as index, sweat is carried out point
Stage regulates and controls, and whole sweat is divided into earlier fermentation, ferment middle and 3 stages of fermentation later stage；At earlier fermentation, thalline
Biomass increases the vigorous stage, and rotating speed is initially set 0rpm, and dissolved oxygen is initially set in 100%, as cell concentration increases, molten
Oxygen can present downward trend, by rotating speed being improved 5rpm at interval of 4 hours, until 50rpm stops so that dissolved oxygen can after declining
Stable at 40%-70%；At ferment middle, in the oil and fat accumulation stage, control dissolved oxygen at 20%-40%；Phase after fermentation, arachidonic
In the acid biosynthesis stage, control dissolved oxygen at 40%-70%.
During the 0-72 hour for fermentation for the described earlier fermentation.
The control of described earlier fermentation dissolved oxygen is realized by air quantity and rotating speed control, and Boiler pressure control is at 1-3VVM(VVM: single
The tolerance being passed through in the volume unit minute of position), rotating speed controls at 0-50 rev/min.
During the 73-120 hour for fermentation for the described ferment middle.
The control of described ferment middle dissolved oxygen is realized by air quantity and rotating speed control, and Boiler pressure control, at 0.5-1.5VVM, turns
Speed controls at 50-200 rev/min.
During the 121-216 hour for fermentation for the described fermentation later stage.
Described fermentation later stage dissolved oxygen control is realized by air quantity and rotating speed control, and Boiler pressure control is in 1-3VVM, rotating speed control
System is at 100-200 rev/min.
The control of described earlier fermentation dissolved oxygen is realized by air quantity and rotating speed control, and Boiler pressure control is in 1-2VVM, rotating speed control
System, at 0-50 rev/min, controls dissolved oxygen between 40%-55%.
The control of described ferment middle dissolved oxygen is realized by air quantity and rotating speed control, and Boiler pressure control, at 0.8-1.2VVM, turns
Speed controls at 80-150 rev/min, controls dissolved oxygen between 20%-35%.
Described fermentation later stage dissolved oxygen control is realized by air quantity and rotating speed control, and Boiler pressure control is at 1.5-2VVM, rotating speed
Control, at 150-200 rev/min, controls dissolved oxygen at 40%-50%.
Mortierella alpine trichoderma strain employed in the present invention is Mortierella alpine trichoderma strain R807(CCTCC M 2012118)
Principle of adjustment and control of the present invention is as follows: fermentation step is divided into three phases (biomass accumulation stage, oil and fat accumulation rank
Section, arachidonic acid biosynthesis stage), and use different modulation process for the different stages, at biomass accumulation
In the stage, seed liquor has just tapped into fermentation tank, and thalline vigor is relatively low more fragile, not resistant to shearing, uses the training method of gas lift pot type,
High dissolved oxygen low sheraing, the beneficially accumulation of Mortierella alpina cell quantity.When entering the oil and fat accumulation stage, biomass reaches certain journey
Degree, single ventilating mode is not enough to form, to zymotic fluid, the effect seethed, coordinates suitable speed of agitator, the gas to zymotic fluid
Liquid mixed effect has well contribution.The phase arachidonic acid biosynthesis stage after fermentation, suitable reduction rotating speed and carrying
High air quantity, beneficially Δ 9 desaturase, Δ 12 desaturase, Δ 6 desaturase and Δ 5 desaturase play a role so that flower
The percentage composition of raw tetraenoic acid rises.
Beneficial effects of the present invention:
(1) more fragile for thalline early stage, the problems such as later stage density is excessive, the preferable and mechanical agitation in conjunction with gas lift tank dissolved oxygen
Tank blade can provide the advantages such as shearing.By the regulation and control to technique, improve fermentation productivity, arachidonic acid in 7.5L tank
Specific yield reach 20.628g/L zymotic fluid, arachidonic production intensity reaches 2.291g/ (L*d).
(2) technological operation that the present invention relates to is easy, beneficially industrialized production.At present in 7m and 25m fermentation
Tank level completes trail run, and obtains good income.
Detailed description of the invention
Below in conjunction with detailed description of the invention, describe the present invention in detail.Should be understood that these embodiments are merely to illustrate this
Bright rather than limit the scope of the present invention.In addition, it is to be understood that after having read the content that the present invention lectures, art technology
The present invention can be made various changes or modifications by personnel, and these equivalent form of values fall within the application appended claims equally and limited
Fixed scope.
Culture medium employed in following example is as follows:
Test tube slant culture medium (PDA slant medium): fresh peeled potatoes 200g, boils rear timing 30min, 4 layers
Filtered through gauze removes solid content, adds glucose 20g, agar 20g, is settled to 1L with distilled water, and pH is natural, 121 DEG C of sterilizings
30min。
Seed culture medium: glucose 30g/L, yeast extract 6g/L, KH2PO4 3g/L, pH are natural, 121 DEG C of sterilizing 30min.
Fermentation medium: glucose 80g/L, yeast extract 11g/L, KH2PO4 3.8g/L, NaNO33.4g/L, MgSO4·
7H2O 0.5g/L, pH are natural, 121 DEG C of sterilizing 30min.
Bacterial strain used in embodiment is: Mortierella alpine trichoderma strain R807(CCTCC M 2012118), it is possible to use its
He is similar to bacterial strain.
Embodiment 1 is based on Dissolved oxygen regulation strategy 7.5L tank Mortierella alpina fermentation producing arachidonic acid oil
1st, the preparation of the activation of bacterial classification and seed liquor: choose Mortierella alpine trichoderma strain R807(CCTCC M 2012118)
For starting strain, the bacterial classification of preservation is accessed in PDA slant medium, cultivate 10 days in incubator under the conditions of 25 DEG C, turn
It is connected to cultivate in constant temperature oscillator equipped with in the 500mL groove bottle of 100mL seed culture medium, inoculum concentration 10%(v/v, 10mL),
Condition of culture is 120rpm, temperature 25 DEG C, incubation time 1-2 days.
2nd, the seed liquor that step 1 obtains is inoculated in 7.5L tank (New Brunswick Scientific, USA), dress
Liquid measure 5L, inoculum concentration is 10%(v/v, 500mL), cultivation temperature 0-120h controls at 28 DEG C, and 121-216h controls at 18 DEG C,
Fermentation is initial, and seed liquor has just tapped into fermentation tank, has one section of laundering period, and initial air quantity is 2VVM, and speed of agitator is initial
Being 50 revs/min, dissolved oxygen initially sets 100%, during the 0-72 hour of fermentation, is earlier fermentation, and Fungal biodiversity increases vigorous
In the stage, after the increase of cell concentration, dissolved oxygen constantly declines, and being controlled by air quantity and rotating speed can after dissolved oxygen declines
It is stable that at 40%-70%, Boiler pressure control is initially set 0rpm at 1-2VVM, rotating speed, at interval of 4 hours, rotating speed is improved 5rpm,
Until 50rpm stops；During the 73-120 hour of fermentation, being ferment middle, this stage is the oil and fat accumulation stage, by air quantity and
Dissolved oxygen is controlled at 20%-40% by rotating speed control, and Boiler pressure control is at 1-1.5VVM, and rotating speed controls at 150-200 rev/min；Sending out
During the 121-216 hour of ferment, being the fermentation later stage, this stage is the arachidonic acid biosynthesis stage, controls dissolved oxygen
40%-70%, Boiler pressure control is at 1-2VVM, and rotating speed controls at 150-200 rev/min.Following table is with the control of different phase separation
It as a example by condition, is shown in Table 1.
Table 1
Note: just completed inoculation when because of Fungal biodiversity relatively low, be difficult to detection, therefore within 0 hour, do not sample detection biomass,
Fat content and arachidonic acid percentage composition.
Fermentation is initial, and seed liquor has just tapped into fermentation tank, has one section of laundering period, and initial air quantity is 2VVM, and speed of agitator is initial
Being 50 revs/min, dissolved oxygen initially sets 100%, and after cell concentration growth, dissolved oxygen constantly declines, and controls about 40%；
When fermented and cultured to 72 is little, Fungal biodiversity increases and slows down, and now intracellular grease starts synthesis, by Boiler pressure control
At 1VVM, speed of agitator controls at 150 revs/min, and dissolved oxygen controls 20%；
Fermented and cultured is to after 120 hours, and now in tank, cell concentration is excessive, and fermentation broth viscosity substantially rises, by rotating speed
Control is at 200 revs/min, and Boiler pressure control gas-liquid mixed effect in 2VVM, control fermentation tank, dissolved oxygen controls about 50%；
When fermented and cultured to 216 is little, fermentation ends, sampling detects the biomass of terminal, fat content and arachidonic
Acid percentage composition.Mortierella alpina dry cell weight, fat content, arachidonic acid percentage composition, arachidonic unit are produced
Amount can reach 56.1g/L, the 53.9%th, 68.22%, 20.628g/L respectively, and arachidonic acid produces intensity and reaches 2.292g/ (L*
d)。
Embodiment 2. is based on Dissolved oxygen regulation strategy 7m tank fermentation Mortierella alpina producing arachidonic acid
1st, the preparation of the activation of bacterial classification and seed liquor: choose Mortierella alpine trichoderma strain R807(CCTCC M 2012118)
For starting strain, the bacterial classification of preservation is accessed in PDA slant medium, cultivate 10 days in incubator under the conditions of 25 DEG C, turn
It is connected to cultivate in constant temperature oscillator equipped with in the 500mL groove bottle of 100mL seed culture medium, inoculum concentration 10%(v/v, 10mL),
Condition of culture is 120rpm, temperature 25 DEG C, and incubation time 1-2 days is prepared as primary seed solution.Then, according to inoculum concentration 10%
(v/v) accessing primary seed solution in 100L seeding tank, condition of culture is 200rpm, temperature 25 DEG C, incubation time 1 day, system
Standby one-tenth secondary seed solution.Then, being all linked in 3000L seeding tank by the secondary seed solution preparing, condition of culture is
100rpm, temperature 25 DEG C, incubation time 1 day, it is prepared as three grades of seed liquor.
2nd, the three grades of seed liquor obtaining step 1 are all linked in 7m fermentation tank, and cultivation temperature 0-120h controls 28
DEG C, 121-216h controls at 18 DEG C, and the control of remaining parameter process is shown in Table 2.The ratio of height to diameter 3:1 of fermentation tank wherein used, by
Nanjing Highke Bioengineering Equipment Co., Ltd. manufactures installs, and Jiangsu Tiankai Biotechnology Co., Ltd. uses and manages, and fills liquid
During measuring 5m, the 0-72 hour of fermentation, being earlier fermentation, Fungal biodiversity increases the vigorous stage, with the increase of cell concentration
After, dissolved oxygen constantly declines, and is controlled after dissolved oxygen declines by air quantity and rotating speed and can stablize at 40%-45%, Boiler pressure control
At 1-2VVM, rotating speed is initially set 0rpm, at interval of 4 hours, rotating speed is improved 5rpm, until 50rpm stops；73-in fermentation
During 120 hours, being ferment middle, this stage is the oil and fat accumulation stage, is controlled by air quantity and rotating speed and controls dissolved oxygen
20%-35%, Boiler pressure control is at 0.8-1.2VVM, and rotating speed controls at 80-150 rev/min；During the 121-216 hour of fermentation,
For fermentation the later stage, this stage is the arachidonic acid biosynthesis stage, controls dissolved oxygen at 40%-50%, and Boiler pressure control is at 1.5-
2VVM, rotating speed controls at 150-200 rev/min.Following table is as a example by the control condition of different phase separation, and control condition is shown in Table
2。
Table 2
Note: just completed inoculation when because of Fungal biodiversity relatively low, be difficult to detection, therefore within 0 hour, do not sample detection biomass,
Fat content and arachidonic acid percentage composition.
Fermentation is initial, and seed liquor has just tapped into fermentation tank, has one section of laundering period, and initial air quantity is 1.5VVM, at the beginning of speed of agitator
Beginning is 0, and every four hours, regulation frequency converter made rotating speed increase by 5 turns, and till reaching 50 revs/min, dissolved oxygen initially sets 100%,
After cell concentration growth, dissolved oxygen constantly declines, and can control about 45%；
When fermented and cultured to 72 is little, Fungal biodiversity increases and slows down, and now intracellular grease starts synthesis, by Boiler pressure control
At 1VVM, speed of agitator controls at 100 revs/min, and dissolved oxygen controls 20%；
Fermented and cultured is to after 120 hours, and now in tank, cell concentration is excessive, and fermentation broth viscosity substantially rises, by rotating speed
Control is at 160 revs/min, and Boiler pressure control gas-liquid mixed effect in 1.7VVM, control fermentation tank, dissolved oxygen controls about 40%；
When fermented and cultured to 216 is little, fermentation ends, sampling detects the biomass of terminal, fat content and arachidonic
Acid percentage composition.Mortierella alpina dry cell weight, fat content, arachidonic acid percentage composition, arachidonic unit are produced
Amount can reach 53.08g/L, the 57.05%th, 62.37%, 18.887g/L respectively, and arachidonic acid produces intensity and reaches 2.099g/
(L*d)。
Embodiment 3. is based on Dissolved oxygen regulation strategy 25m tank fermentation Mortierella alpina producing arachidonic acid
1st, the preparation of the activation of bacterial classification and seed liquor: choose Mortierella alpine trichoderma strain R807(CCTCC M 2012118)
For starting strain, the bacterial classification of preservation is accessed in PDA slant medium, cultivate 10 days in incubator under the conditions of 25 DEG C, turn
It is connected to cultivate in constant temperature oscillator equipped with in the 500mL groove bottle of 100mL seed culture medium, inoculum concentration 10%(v/v, 10mL),
Condition of culture is 120rpm, temperature 25 DEG C, and incubation time 1-2 days is prepared as primary seed solution.Then, according to inoculum concentration 10%
(v/v) accessing primary seed solution in 150L seeding tank, condition of culture is 200rpm, temperature 25 DEG C, incubation time 1 day, system
Standby one-tenth secondary seed solution.Then, being all linked in 5000L seeding tank by the secondary seed solution preparing, condition of culture is
100rpm, temperature 25 DEG C, incubation time 1 day, it is prepared as three grades of seed liquor.
2nd, the three grades of seed liquor obtaining step 1 are all linked in 25m fermentation tank, and cultivation temperature 0-120h controls
28 DEG C, 121-216h controls at 18 DEG C, during the 0-72 hour of fermentation, is earlier fermentation, and Fungal biodiversity increases the vigorous stage,
After the increase of cell concentration, dissolved oxygen constantly declines, and is controlled after dissolved oxygen declines can be stablized by air quantity and rotating speed
At 40%-55%, Boiler pressure control is initially set 0rpm at 1-2VVM, rotating speed, at interval of 4 hours, rotating speed is improved 5rpm, until
50rpm stops；During the 73-120 hour of fermentation, being ferment middle, this stage is the oil and fat accumulation stage, by air quantity and rotating speed
Dissolved oxygen is controlled at 20%-35% by control, and Boiler pressure control is at 0.8-1.2VVM, and rotating speed controls at 80-150 rev/min；In fermentation
During 121-216 hour, being the fermentation later stage, this stage is the arachidonic acid biosynthesis stage, controls dissolved oxygen at 40%-
50%, Boiler pressure control is at 1.5-2VVM, and rotating speed controls at 150-200 rev/min.Following table is with the control strip of different phase separation
As a example by part, the control of parameter process is shown in Table 3.Fermentation tank ratio of height to diameter 2.5:1 wherein used, is set by Chaoyang, yiyang, hunan city lightization
Standby factory manufactures installs, and Jiangsu Tiankai Biotechnology Co., Ltd. uses and manages, liquid amount 21m, and the control of zymotechnique is shown in Table
3:
Table 3
Note: just completed inoculation when because of Fungal biodiversity relatively low, be difficult to detection, therefore within 0 hour, do not sample detection biomass,
Fat content and arachidonic acid percentage composition.
Fermentation is initial, and seed liquor has just tapped into fermentation tank, has one section of laundering period, and initial air quantity is 1.5VVM, at the beginning of speed of agitator
Beginning is 0, and every four hours, regulation frequency converter made rotating speed increase by 5 turns, and till reaching 50 revs/min, dissolved oxygen initially sets 100%,
After cell concentration growth, dissolved oxygen constantly declines, and can control about 60%；
When fermented and cultured to 72 is little, Fungal biodiversity increases and slows down, and now intracellular grease starts synthesis, by Boiler pressure control
At 1VVM, speed of agitator controls at 80 revs/min, and dissolved oxygen can control 35%；
Fermented and cultured is to after 120 hours, and now in tank, cell concentration is excessive, and fermentation broth viscosity substantially rises, by rotating speed
Control is at 150 revs/min, and Boiler pressure control gas-liquid mixed effect in 1.7VVM, control fermentation tank, dissolved oxygen controls about 40%；
When fermented and cultured to 216 is little, fermentation ends, sampling detects the biomass of terminal, fat content and arachidonic
Acid percentage composition.Mortierella alpina dry cell weight, fat content, arachidonic acid percentage composition, arachidonic unit are produced
Amount can reach 49g/L, the 50.42%th, 63.86%, 15.777g/L respectively, and arachidonic acid produces intensity and reaches 1.753g/ (L*
d)。
Comprehensive above example is it can be seen that utilize of the present invention producing based on the fermentation of Dissolved oxygen regulation Mortierella alpina to spend
The method of raw tetraenoic acid grease so that biomass, fat content, arachidonic acid percentage composition, and arachidonic acid unit
The many aspects such as yield and production intensity are all improved largely.
Claims (4)
1. the method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil, by Mortierella alpine trichoderma strain warp
Cross test tube slant culture medium and Shake flask medium amplifies cultivation step by step, prepare seed liquor, be then inoculated in fermentation tank and carry out sending out
Ferment cultivate, it is characterised in that: during the fermentation, with control Mortierella alpina fermentation form as target, with dissolved oxygen parameter of fermenting
For index, sweat is regulated and controled stage by stage, whole sweat is divided into earlier fermentation, ferment middle and fermentation later stage
3 stages；Rotating speed is initially set 0rpm, and dissolved oxygen is initially set in 100%, and as cell concentration increases, dissolved oxygen can present decline
Trend, by first stepping up rotating speed, and combines the method for operating improving air quantity, at earlier fermentation so that dissolved oxygen is steady after declining
It is scheduled on 40%-70%；At ferment middle, control dissolved oxygen at 20%-40%；Phase after fermentation, control dissolved oxygen at 40%-70%；Institute
During stating the 0-72 hour for fermentation for the earlier fermentation；The control of described earlier fermentation dissolved oxygen is realized by air quantity and rotating speed control,
Boiler pressure control is at 1-3VVM, and rotating speed controls at 0-50 rev/min；During the 73-120 hour for fermentation for the described ferment middle；Institute
Stating the control of ferment middle dissolved oxygen to be realized by air quantity and rotating speed control, Boiler pressure control controls at 50-at 0.5-1.5VVM, rotating speed
200 revs/min；During the 121-216 hour for fermentation for the described fermentation later stage；Described fermentation later stage dissolved oxygen control by air quantity and
Rotating speed control realizes, Boiler pressure control is at 1-3VVM, and rotating speed controls at 100-200 rev/min.
2. the method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil according to claim 1, its
It is characterised by: the control of described earlier fermentation dissolved oxygen is realized by air quantity and rotating speed control, and Boiler pressure control is in 1-2VVM, rotating speed control
System, at 0-50 rev/min, controls dissolved oxygen between 40%-55%.
3. the method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil according to claim 1, its
It is characterised by: the control of described ferment middle dissolved oxygen is realized by air quantity and rotating speed control, and Boiler pressure control, at 0.8-1.2VVM, turns
Speed controls at 80-150 rev/min, controls dissolved oxygen between 20%-35%.
4. the method based on Dissolved oxygen regulation Mortierella alpina fermentation producing arachidonic acid oil according to claim 1, its
It is characterised by: described fermentation later stage dissolved oxygen control is realized by air quantity and rotating speed control, and Boiler pressure control is at 1.5-2VVM, rotating speed
Control, at 150-200 rev/min, controls dissolved oxygen at 40%-50%.
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