CN107287125A - A kind of cultural method of chlorella pyrenoidosa - Google Patents
A kind of cultural method of chlorella pyrenoidosa Download PDFInfo
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- CN107287125A CN107287125A CN201710733936.9A CN201710733936A CN107287125A CN 107287125 A CN107287125 A CN 107287125A CN 201710733936 A CN201710733936 A CN 201710733936A CN 107287125 A CN107287125 A CN 107287125A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/12—Unicellular algae; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
Abstract
The present invention provides a kind of beneficial to raising chlorella pyrenoidosa growth rate, increase cell density, promote the cultural method of oil and fat accumulation, be the mixed fermentation bacterium solution for red bacterium category (Rhodobacter sp.) and Rhodopseudomonas (Rhodopseudomonas sp.) bacterium that inactivation is with the addition of in the culture medium of culture chlorella pyrenoidosa.The method of the present invention can make frustule fast-growth, and the chlorella pyrenoidosa cell density turned out is big, especially because with the addition of mixed bacteria liquid; so as to promote frustule fast breeding; so that frustule concentration increases, fat content substantially increases, suitable for the production of high-volume pilot scale culture.
Description
Technical field
The invention belongs to algae culture technical field, and in particular to a kind of cultural method of chlorella pyrenoidosa.
Background technology
Chlorella pyrenoidosa (Chlorella pyrenoidosa) belongs to Chlorophyta, Chroococcales, Chlorella, is this
Unique species with pyrenoids in platymiscium.Contain abundant protein, polysaccharide, unsaturated lipid in chlorella pyrenoidosa cell
Fat acid, dietary fiber, vitamin and trace element etc., are the mankind's excellent health foods and water with very high nutritive value
Produce breeding bait.New resource food is approved as by Ministry of Health of the People's Republic of China.Chlorella pyrenoidosa can be again as a class
Raw energy organism matter can accumulate grease under the condition of culture of light autotrophy and heterotrophism, effectively using solar energy and can have function to enter
Row fast-growth and accumulation grease, the grease of chlorella pyrenoidosa can be biological as the renewable sources of energy, for making biodiesel
Diesel oil has very big potentiality on substitute fossil fuels diesel oil.But current chlorella pyrenoidosa pilot scale culture has life
Long slow, biomass is low, the problem of fat content is low, is unfavorable for large-scale culture.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of thin beneficial to raising chlorella pyrenoidosa growth rate, increase
Born of the same parents' density, promotes the cultural method of oil and fat accumulation, so as to improve culture efficiency, is particularly suitable for the pyrenoids bead of pilot scale culture
Algae cultural method.
The cultural method of chlorella pyrenoidosa provided by the present invention, is added in the culture medium of culture chlorella pyrenoidosa
Red bacterium category (Rhodobacter sp.) and Rhodopseudomonas (Rhodopseudomonas sp.) bacterium of inactivation is added
Zymocyte liquid；
Described red bacterium category (Rhodobacter sp.), one kind of embodiment is preferably Rhodobacter capsulatus
(Rhodobacter capsulatus)；
Described Rhodopseudomonas (Rhodopseudomonas sp.) bacterium, one kind of embodiment is preferably marsh
Red pseudomonas (Rhodopseudomonas palustris)；
The culture medium of described zymocyte liquid, concrete composition is as follows：NH4Cl 1g/L、K2HPO4 0.5g/L、MgCl2
0.2g/L, dusty yeast 0.5g/L, sodium acetate 4g/L.Fermentation temperature is 30 DEG C, 24 hours continuous lights, and intensity is 10000Lux,
Fermentation time is 3 days, every kind of microbionation concentration 1 × 107Individual/mL.
Used chlorella pyrenoidosa culture medium, specific composition is as follows：Peptone 300-500mg/L, K2HPO4 16-
21mg/L, MgSO4·7H2O 60-90mg/L, CaCl2·2H2O 30-42mg/L, ferric citrate 8-16mg/L, NaHCO3
10-30mg/L, trace element solution 0.5-1.5ml/L.
Auximone and vitamin solution can also be added in above-mentioned culture medium.
Described auximone is heteroauxin, and vitamin solution is vitamin B2Solution.
More specifically, described cultural method, its temperature selected when cultivating is 30 DEG C, 24 hours continuous lights, intensity
For 10000Lux.
The method of the present invention can make frustule fast-growth, and the chlorella pyrenoidosa cell density turned out is big, especially
It is due to that with the addition of mixed bacteria liquid, so as to promote frustule fast breeding so that frustule concentration increases, fat content substantially increases
Plus, suitable for the production of high-volume pilot scale culture.
Brief description of the drawings
The influence figure of Fig. 1 different culture medias concentration and its flora to chlorella pyrenoidosa fat content.
Embodiment
In order to which technical characteristic, purpose and beneficial effect to the present invention are more clearly understood from, now to the skill of the present invention
Art scheme carry out it is described further below, but it is not intended that to the present invention can practical range restriction.
Raw material involved in the present invention and method are described as follows below：
It is higher that various nutritive salt (in addition to auximone and vitamin solution) used in the present invention can first be made into concentration
Mother liquor, generally mother liquor be configured to concentration 1000 times, prepare mother liquor (1)-(7) number when, medicine is prepared respectively, is deposited respectively
Put；(8) mixed storage after the medicine in number trace element is prepared according to quantity.After various mother liquors have been matched somebody with somebody, added respectively with Brown Glass Brown glass bottles and jars only
Lid storage, it is labelled, indicate storage liquid title, prepare multiple, date etc., be stored in cold compartment of refrigerator.Prepare working solution
During culture medium, storage liquid is first gently shaken, if it find that having precipitation, suspension or being polluted by Institute of Micro-biology, the storage liquid needs
Match somebody with somebody again.Finally according to the culture volume of preparation with liquid-transfering gun from various storage liquid required consumption, Ensure Liquid are taken out respectively
Need constantly to add water and stir during salt, and various nutritive salt are added by the order provided in formula, to avoid chemistry
Reaction, plus distilled water diluting is to required concentration.The culture medium prepared is adjusted with pH meter to 7.0, packing, after wrapping with
0.103MPa, 121 DEG C, autoclaving 20min.
Grease is extracted and assay：Dry algae powder M1 is weighed, it is 1 to add certain volume ratio：2 chloroform：Methanol solution, shakes
Mixing is swung, is extracted with ultrasonic disruption machine, the time is 20-30min；5min is centrifuged with 3000r/min again, takes chloroform layer molten
Liquid repeats to extract after grease 3 times in the glass centrifuge tube for weighing M2 in advance, the chloroform layer of collection is dried up in nitrogen and
Dried in vacuum desiccator, weigh to constant weight M3, carry out weighing calculating
Total lipid content is calculated as follows：Total fat (%)=100 × (M3-M2)/M1.
Nile red decoration method：The algae solution of certain volume exponential phase middle and later periods is taken, 8 000r/min centrifugation 5min are removed
Supernatant, frustule precipitation is washed twice with phosphate buffer, uses volume fraction to be resuspended for 20% dimethyl sulphoxide aqueous solution
Frustule, makes algae solution OD5400.8,40 DEG C of water-bath 20min of value, adding 15 μ L Nile red dyes by 1mL algae solutions, (mass concentration is
0.1mg/mL acetone solns), dyeing 5min, excitation wavelength 480nm are mixed, determines it in the fluorescence intensity of 575nm wavelength to sieve
Look into the size of algae kind total lipid content.
With reference to specific embodiment, the present invention will be described in detail.
Embodiment 1：Screening and structure of community of the chlorella pyrenoidosa with mixed bacterial cogeneration system are identified
The water sample extracted from the municipal sewage plant biochemistry pool activated sludge of Harbin, loads sterile sampling bag and takes back experiment
Room, microscope inspection observation finds that sample microbial and alga cells are more, corresponding microalgae and microalgae association flora is carried out rich
Collection, separation screening.Raw water sample is delivered and continuously cultivated in simulation biochemistry pool reactor, reactor is placed under natural lighting, with
Synthetic municipal wastewater well prepared in advance is mixed, long-time Air Exposure, and oxygen content is maintained at 6-8mg/L, 32 DEG C of cultures, pH
7.5COD is maintained at 400mg/L or so, TN 50mg/L or so, TP 10mg/L or so.Simulated by the BG11 culture mediums of improvement
Municipal sewage, it is following (mg/L) that it simulates sewage composition：Peptone 400, potassium dihydrogen phosphate 18.5, epsom salt 75.0, two
Water calcium chloride 36, ferric citrate 12, sodium acid carbonate 20 finally adds trace element 1mL.Wherein trace element formula is (mg/
L)：H3BO31660, MnCl2·4H2O 1860, ZnSO4·7H2O 220, Na2MoO4·2H2O 21, CuSO4·5H2O 80, Co
(NO3)2·6H2O 50, NiCl250, KI 30.
According to testing result, when the COD in cultivating system is reduced to 300mg/L, addition 100mg/L glucose is used as benefit
Fill carbon source.EGR is set in aeration tank, and 1.5L/min, equivalent 5h can be with circulation primary water sample so that water body flows back and mixes
It is even, after the culture of continuous aeration pond is more than 20d, obtain one it is activity stabilized, the active bacterium algae that can be resistant to certain organic concentrations is common
Raw system (as described below), number of viable maintains 109Individual/more than mL, microalgae quantity maintains 107Individual/mL.At the place of waste water
During reason, water sample starting COD is 100mg/L, biochemical treatment using the method that steps up waste water ratio, make functional flora by
Step adapts to high concentration municipal wastewater environment.The simulated wastewater of first generation domestication system addition 1/4, the distilled water of supplement 3/4, inoculation
Flora, inoculum concentration 20% (v/v), 30 DEG C of shaking tables, 160rpm shaken cultivations.COD degradation rate is 40.25% after 2d, continues to cultivate
COD keeps stable and no longer declined.It is forwarded in second generation domestication culture medium, improves waste water ratio.The simulated wastewater of 50% volume
Add the distilled water of 50% volume, inoculum concentration 20%, 30 DEG C of shaking table cultures.Detect that COD is reduced from initial 210.5mg/L after 2d
To 68.75mg/L, degradation rate is 67.34%, and so on, into forth generation domestication system, all simulation municipal administrations of culture medium
Sewage, active microalgae symbiosis flora is as shown in table 1 to the Biochemical Treatment of waste water.COD is down to 37.53mg/L or so in sewage
And keeping activity stabilized, COD clearances are 91.00%, close with raw sewage factory biochemistry pool effluent quality, illustrate to have obtained one group
There is remarkable result in terms of stable helotism system, its COD in reduction system.
The helotism system acclimation sewage effect table of table 1
The mixing microalgae flora obtained using pellet fraction circumfluence method to enrichment is screened, is identified and isolated from, and is utilized
Form and Molecular tools carry out COMMUNITY STRUCTURE to the helotism system, find to be primarily present in bacteria flora it is following into
Member：Red bacterium category (Rhodobacter sp.), Rhodopseudomonas (Rhodopseudomonas sp.), bacillus genus
(Paenibacillus sp.), enterobacteria (Enterobacter sp.), color salt bacillus (Chromohalobacter sp.),
Chryseobacterium sp (Chryseobacterium sp.), sum to more than 80%.Wherein red bacterium category (Rhodobacter sp.) and
Rhodopseudomonas (Rhodopseudomonas sp.) ratio highest, respectively reaches 21% and 19%, next to that class gemma bar
Pseudomonas (Paenibacillus sp.) and enterobacteria (Enterobacter sp.) ratio are 11%, color salt bacillus
(Chromohalobacter sp.) and Chryseobacterium sp (Chryseobacterium sp.) account for 10% and 9% respectively.It is active thin
In bacteria microorganism group, red bacterium category (Rhodobacter sp.), Rhodopseudomonas (Rhodopseudomonas sp.) and
Bacillus genus (Paenibacillus sp.) is widely present in nature and major sewage treatment plant's biochemistry pools, autotrophy
Or can be grown under the conditions of different oxygen, participate in the conversion of the materials such as carbon, nitrogen and P elements and circulate.Enterobacteria (Enterobacter)
And color salt bacillus (Chromohalobacter) has certain halophagia, with certain degradation of organic substances function, in all kinds of dirts
The microorganism being widely present in water.There is the function bacterium of municipal administration and chemical wastewater treatment in Chryseobacterium sp (Chryseobacterium)
In group, while the organic matter such as aromatic hydrocarbons and benzene class that can degrade.
By the above method and means, it is found that microalgae main species include two kinds of microalgaes：Chlorella (Chlorella sp.)
And grid algae (Scenedesmus sp.), wherein chlorella account for sum 90%.Two plants of microalgaes are entered by Nile red decoration method
Row fat content is determined, it is found that chlorella fat content reaches 21.25%, total fat yield reaches 113.8mg/ (Ld), grid algae
Fat content reaches 18.76%, and total fat yield reaches 87.8mg/ (Ld).The present invention passes through 16S rRNA equimolecular biology
Means are identified that it is chlorella pyrenoidosa to determine the oil-rich microalgae chlorella (Chlorella sp.) in the inventive method
(Chlorella pyrenoidosa)。
Embodiment 2：Compound flora and activity checking
Simulate the red bacterium category (Rhodobacter sp.) of photosynthetic bacteria microbiologic population in embodiment 1 and red pseudomonas
Belong to the proportion of composing of (Rhodopseudomonas sp.), and chlorella pyrenoidosa (Chlorella pyrenoidosa) is carried out
Compounding.Wherein red bacterium category (Rhodobacter sp.) plants Rhodobacter capsulatus (Rhodobacter from specific
Capsulatus), Rhodopseudomonas (Rhodopseudomonas sp.) is planted as damp red pseudomonas from specific
(Rhodopseudomonas palustris).Wherein single bacterium (algae) is activated with LB culture mediums respectively, cell is adjusted after activation dense
Degree, and chlorella pyrenoidosa addition concentration is more than two kinds of photosynthetic bacteria concentration, two orders of magnitude, compounds obtained microorganism bacterium algae
System is according to fat content detection method described in embodiment, and analysis compounding flora is to chlorella pyrenoidosa fat content
Influence.It is 35.43% that culture, which terminates rear chlorella pyrenoidosa fat content, and growth is stable, and algae cell density reaches 5 × 107
Individual/mL, total fat yield reaches 133.9mg/ (Ld).The above results show that the composite flora prepared demonstrates work(in embodiment 1
Energy flora grows to the facilitation of chlorella pyrenoidosa Lipid-producing while two kinds of single bacteriums of explanation compound obtained flora for algae
The influence of density and fat content and COD of sewage removal efficiency is better than the helotism system screened in embodiment 1, respectively
Improve 400%, 66.73% and 7.79%.
Embodiment 3：Different culture media concentration and its flora concentration are to chlorella pyrenoidosa growth and the optimization of fat content
The chlorella pyrenoidosa being formulated as follows is configured with sterilized water and optimizes wild Oryza species：Peptone 300mg/L, KH2PO4
16mg/L, MgSO4·7H2O 60mg/L, CaCl2·2H2O 30mg/L, ferric citrate 8mg/L, NaHCO310mg/L, it is micro
Element Solution 0.5ml/L；Also include auximone heteroauxin 1.0mg/L and vitamin B2Solution 0.5mg/L.Including pod membrane
Red bacterium (Rhodobacter capsulatus), red damp red pseudomonas (Rhodopseudomonas palustris) it is mixed
Microflora fermentation liquid 10mL is closed, two of which strain inoculation proportioning is 1:1.Its mixed bacterial zymotic fluid concrete composition is as follows：NH4Cl
1g/L、K2HPO4 0.5g/L、MgCl20.2g/L, dusty yeast 0.5g/L, sodium acetate 4g/L.Fermentation temperature is 30 DEG C, 24 hours
Continuous light, intensity is 10000Lux, and fermentation time is 3 days, inoculative proportion 1：1, every kind of microbionation concentration 1 × 107Individual/
mL.The chlorella pyrenoidosa in growth period of taking the logarithm is inoculated into the culture medium and cultivated, 24 hours continuous lights, and intensity is about
10000Lux, cultivation temperature controls 30 degrees centigrades, shakes twice manually daily, prevents from precipitating adherent, periodic logging culture
The cell concentration of chlorella pyrenoidosa in base.BG-11 culture mediums are used in addition as control medium, not additional Mixed Microbes mass-sending
Zymotic fluid and special dietary material are used as control experiment group.Each experimental group algae density value is measured in the exponential phase middle and later periods, in detail
It is shown in Table 2；Chlorella pyrenoidosa fat content is measured simultaneously, Fig. 1 is referred to.
The different flora matched proportion densities of table 2 and nutrient concentration are to chlorella pyrenoidosa growth effect contrast table
Embodiment 4：Different culture media concentration and its flora concentration are to chlorella pyrenoidosa growth and the optimization of fat content
The chlorella pyrenoidosa being formulated as follows is configured with sterilized water and optimizes wild Oryza species：Peptone 300mg/L, KH2PO4
16mg/L, MgSO4·7H2O 60mg/L, CaCl2·2H2O 30mg/L, ferric citrate 8mg/L, NaHCO310mg/L, it is micro
Element Solution 0.5ml/L；Also include auximone heteroauxin 1.0mg/L and vitamin B2Solution 0.5mg/L.Including red thin
Pseudomonas (Rhodobacter sp.) and Rhodopseudomonas (Rhodopseudomonas sp.) mixed bacterial zymotic fluid 30mL,
Mixed bacterial zymotic fluid group is into, fermentation condition and bacterium algae inoculative proportion be the same as Example 3.Take the logarithm the chlorella pyrenoidosa in growth period
It is inoculated into the culture medium and is cultivated, 24 hours continuous lights, intensity is about 10000Lux, cultivation temperature controls 30 degrees Celsius
Left and right, shakes twice manually daily, prevents from precipitating adherent, the cell concentration of chlorella pyrenoidosa in periodic logging culture medium.Separately
Outer to use BG-11 culture mediums as control medium, not additional mixed bacterial zymotic fluid and special dietary material are used as control
Experimental group.Each experimental group algae density value is measured in the exponential phase middle and later periods, table 3 is referred to；Simultaneously to chlorella pyrenoidosa grease
Content is measured, and refers to Fig. 1.
The different flora matched proportion densities of table 3 and nutrient concentration are to chlorella pyrenoidosa growth effect contrast table
Embodiment 5：Different culture media concentration and its flora concentration are to chlorella pyrenoidosa growth and the optimization of fat content
The chlorella pyrenoidosa being formulated as follows is configured with sterilized water and optimizes wild Oryza species：Peptone 500mg/L, KH2PO4
21mg/L, MgSO4·7H2O 90mg/L, CaCl2·2H2O 42mg/L, ferric citrate 16mg/L, NaHCO330mg/L, it is micro-
Secondary element solution 1.5ml/L；Also include auximone heteroauxin 5.0mg/L and vitamin B2Solution 1.0mg/L.Including red
Bacterium belongs to (Rhodobacter sp.) and Rhodopseudomonas (Rhodopseudomonas sp.) mixed bacterial zymotic fluid
10mL, mixed bacterial zymotic fluid group is into, fermentation condition and bacterium algae inoculative proportion be the same as Example 3.Take the logarithm the pyrenoids in growth period
Chlorella is inoculated into the culture medium and cultivated, 24 hours continuous lights, and intensity is about 10000Lux, cultivation temperature control 30
Degrees centigrade, shakes twice manually daily, prevents that precipitation is adherent, the cell of chlorella pyrenoidosa is dense in periodic logging culture medium
Degree.BG-11 culture mediums are used as control medium, not additional mixed bacterial zymotic fluid and special dietary material in addition.
The exponential phase middle and later periods measures each experimental group algae density value, refers to table 4；Chlorella pyrenoidosa fat content is surveyed simultaneously
It is fixed, refer to Fig. 1.
The different flora matched proportion densities of table 4 and nutrient concentration are to chlorella pyrenoidosa growth effect contrast table
Embodiment 6：Different culture media concentration and its flora concentration are to chlorella pyrenoidosa growth and the optimization of fat content
The chlorella pyrenoidosa being formulated as follows is configured with sterilized water and optimizes wild Oryza species：Peptone 500mg/L, KH2PO4
21mg/L, MgSO4·7H2O 90mg/L, CaCl2·2H2O 42mg/L, ferric citrate 16mg/L, NaHCO330mg/L, it is micro-
Secondary element solution 1.5ml/L；Also include auximone heteroauxin 5.0mg/L and vitamin B2Solution 1.0mg/L.Including red
Bacterium belongs to (Rhodobacter sp.) and Rhodopseudomonas (Rhodopseudomonas sp.) mixed bacterial zymotic fluid
30mL, mixed bacterial zymotic fluid group is into, fermentation condition and bacterium algae inoculative proportion be the same as Example 3.Take the logarithm the pyrenoids in growth period
Chlorella is inoculated into the culture medium and cultivated, 24 hours continuous lights, and intensity is about 10000Lux, cultivation temperature control 30
Degrees centigrade, shakes twice manually daily, prevents that precipitation is adherent, the cell of chlorella pyrenoidosa is dense in periodic logging culture medium
Degree.BG-11 culture mediums are used as control medium, not additional mixed bacterial zymotic fluid and special dietary material in addition.
The exponential phase middle and later periods measures each experimental group algae density value, refers to table 5；Chlorella pyrenoidosa fat content is surveyed simultaneously
It is fixed, refer to Fig. 1.
The different flora matched proportion densities of table 5 and nutrient concentration are to chlorella pyrenoidosa growth effect contrast table
Find out from result above, cultivated using cultural method provided by the present invention and culture medium ratio using independent BG-11
Base rapidly can breed chlorella pyrenoidosa, chlorella pyrenoidosa is entered increased logarithmic phase in advance, when shortening its culture
Between.It will be seen from figure 1 that chlorella pyrenoidosa fat content contrast control medium BG- under each embodiment optimal culture conditions
11 have the raising of conspicuousness, thus illustrate that the chlorella pyrenoidosa cultural method that the present invention is provided has obvious culture advantage,
The more conducively fast-growth of frustule, while promoting fat content to improve.
Schematical embodiment of the invention is the foregoing is only, the scope of the present invention is not limited to.It is any
Those skilled in the art, made equivalent variations and modification on the premise of the design of the present invention and principle is not departed from,
The scope of protection of the invention should be belonged to.
Claims (10)
1. a kind of cultural method of chlorella pyrenoidosa, it is characterised in that described cultural method is in culture pyrenoids bead
The red bacterium category (Rhodobacter sp.) and Rhodopseudomonas of inactivation are with the addition of in the culture medium of algae
The zymocyte liquid of (Rhodopseudomonas sp.) bacterium.
2. the method as described in claim 1, it is characterised in that described red bacterium belongs to (Rhodobacter sp.) bacterium and is
For Rhodobacter capsulatus (Rhodobacter capsulatus).
3. the method as described in claim 1, it is characterised in that described Rhodopseudomonas (Rhodopseudomonas
Sp.) bacterium, is Rhodopseudomonas palustris (Rhodopseudomonas palustris).
4. the method as described in claim 1, it is characterised in that described zymocyte liquid, its culture medium for using of fermenting is matched somebody with somebody
Than as follows：NH4Cl 1g/L、K2HPO4 0.5g/L、MgCl20.2g/L, dusty yeast 0.5g/L, sodium acetate 4g/L.
5. the method as described in claim 1, it is characterised in that described culture medium, its proportioning is as follows：Peptone 300-
500mg/L, K2HPO416-21mg/L, MgSO4·7H2O 60-90mg/L, CaCl2·2H2O 30-42mg/L, ferric citrate
8-16mg/L, NaHCO310-30mg/L, trace element solution 0.5-1.5ml/L.
6. the method as described in claim 1 or 5, it is characterised in that in described culture medium added with auximone and/or
Vitamin solution.
7. method as claimed in claim 6, it is characterised in that described auximone is heteroauxin, vitamin solution
For vitamin B2Solution.
8. the method as described in claim 1, it is characterised in that cultivation temperature during described culture is 30 DEG C, is held within 24 hours
Continuous illumination, intensity is 10000Lux.
9. a kind of culture medium for being used to cultivate chlorella pyrenoidosa, it is characterised in that with the addition of inactivation in described culture medium
Red bacterium belongs to the zymocyte liquid of (Rhodobacter sp.) and Rhodopseudomonas (Rhodopseudomonas sp.) bacterium.
10. culture medium as claimed in claim 9, it is characterised in that in described culture medium also added with auximone and/
Or vitamin solution.
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| CN108587920A (en) * | 2018-07-20 | 2018-09-28 | 中国科学院武汉植物园 | A method of utilizing acetic acid/sodium acetate mixotrophic cultivation microalgae |
| CN110144368A (en) * | 2019-06-20 | 2019-08-20 | 哈尔滨工业大学 | A method of it maintains persistently to produce hydrogen after chlorella cells are dead |
| CN110157621A (en) * | 2019-05-07 | 2019-08-23 | 天津科技大学 | A kind of preparation method of the highly enriched long-acting preservative agent of microalgae living cells |
| CN112457994A (en) * | 2020-11-27 | 2021-03-09 | 齐鲁工业大学 | Method for promoting growth of chlorella pyrenoidosa by using volatile fatty acid |
| CN113512567A (en) * | 2021-06-29 | 2021-10-19 | 安徽师范大学 | Method for improving oil yield of microalgae in photoheterotrophic system |
| CN116555039A (en) * | 2023-05-13 | 2023-08-08 | 华南理工大学 | Quick culture method of high-quality and high-biomass chlorella pyrenoidosa |
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| CN110157621A (en) * | 2019-05-07 | 2019-08-23 | 天津科技大学 | A kind of preparation method of the highly enriched long-acting preservative agent of microalgae living cells |
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