CN107446822A - A kind of cultural method of rich oil scenedesmus obliquus - Google Patents
A kind of cultural method of rich oil scenedesmus obliquus Download PDFInfo
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Abstract
The invention provides a kind of rich oil scenedesmus obliquus growth rate, increase cell density, the cultural method for shortening incubation time and promotion oil and fat accumulation, is that with the addition of the biological flocculant zymocyte liquid containing a variety of compound flcos producing bacterias in the culture medium of culture scenedesmus obliquus.The method of the present invention can make frustule fast-growth; the scenedesmus obliquus cell density turned out is big; especially because it with the addition of biological flocculant zymotic fluid; so as to promote frustule fast breeding; so that frustule concentration increases, shorten incubation time, further reduce toxigenic capacity; fat content substantially increases, and is produced suitable for 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 scenedesmus obliquus.
Background technology
With becoming increasingly conspicuous for human society shortage of resources and environmental problem, the whole world is faced with energy shortage and ecological ring
The crisis that border is destroyed.Therefore, seeking a kind of green reproducible new energy turns into the section of countries in the world scientist's common concern
Knowledge is inscribed.Biodiesel is clean environment friendly regenerative resource, but because its cost of raw material is higher, current biological bavin
The price of oil remains above conventional diesel.And in numerous biodiesel raw materials, microalgae has more advantages, and microalgae is made with terrestrial
Thing is compared, and is had the characteristics that biomass is big, growth cycle is short, lipid content is high, is not take up arable land, it has also become bio-fuel-oil raw material
First choice.
Scenedesmus obliquus (Scenedesmus obliquas) is a kind of common freshwater microalgae, platymiscium circle, and Chlorophyta is green
Ball Cutleriales, Shan Zao sections, Scenedesmus (Zhao Xinyu, Liao Limin, Cheng Jie, wait urban domestic wastewater resources of the based on scenedesmus obliquus culture
Change research on utilization [J] fwaater resources protections, 2016,32 (3):44‐49.).In warm area, it is that one kind is distributed widely in lake
Fresh water monoplast green alga in the hydrostatic water body such as pool, reservoir, pond, marsh, because strong with easily survival, fertility, environment is resistance to
By property it is strong the features such as.Meanwhile by autotrophy or Heterotrophic culture scenedesmus obliquus, its condition of culture is controlled, alga cells can be a large amount of
Grease is accumulated, scenedesmus obliquus is one of hitherto known oil content highest algae kind, and production biodiesel is converted using microalgae grease,
Raw material as production biodiesel (Ji Xiang, Cheng Jie, Liao Limin, waits .1 strains scenedesmus obliquus to expand optimization [J] of condition of culture
Jiangsu's agriculture science, 2015,43 (10):303‐305.).Higher astaxanthin can be accumulated in scenedesmus obliquus body, can also be made
(Ling Shanfeng, Cai Fuhuan, Liu Yanwen .3.5mg/L Induced by Salicylic Acid scenedesmus obliquus is waited to accumulate for animal feed or food additives etc.
Molecule mechanism [J] Jiangsu's agriculture science of tired astaxanthin, 2016,44 (4):287‐290.).Therefore, the economic valency of scenedesmus obliquus
Value is higher, has preferable development prospect in the field such as the energy and food.
But subject matter existing for current scenedesmus obliquus pilot scale culture is slow-growing, and biomass is low, and in routine
It is vulnerable to the pollution of alien species in cultivating system, causes the collapse of cultivating system, it 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 close beneficial to scenedesmus obliquus growth rate, increase cell is improved
The scenedesmus obliquus culture medium of degree, so as to improve culture efficiency and culture systems stability, particularly suitable for the oblique of pilot scale culture
Raw grid algae culture medium.
The cultural method of scenedesmus obliquus provided by the present invention, be culture scenedesmus obliquus culture medium in the addition of it is compound
Type biological flocculant zymotic fluid, described zymotic fluid are by Klebsiella (Klebsiella sp.)；Rhizobium
Prepared by the fermentation of (Rhizobium sp.) and bacillus (Bacillus sp.).
Described Klebsiella (Klebsiella sp.), one kind of embodiment is preferably Klebsiella
(Klebsiella variicola)；
Described rhizobium (Rhizobium sp.) bacterium, one kind of embodiment is preferably radiation rhizobium
(Rhizobium radiobacter)；
Described bacillus (Bacillus sp.) bacterium, a kind of preferably spherical bacillus of embodiment
(Bacillus sphaeicus)。
Described zymocyte liquid, the fermentation medium concrete composition that it is used are as follows：Glucose 10g/ml, K2HPO4 5g/
L, MgSO4·7H2O 0.2g/L, urea 0.5g/L, H2PO42g/L, NaCl 0.1g/L, yeast extract 0.5g/L, pH 7.5.
Described scenedesmus obliquus culture medium, a kind of its specific formula is as follows：Glucose 10000-150000mg/L,
NaNO3150-250mg/L, MgSO4·7H2O 50-75mg/L, K2HPO450-175mg/L, MnCl2·4H2O 1.00-
1.44mg/L, Co (NO3)2·6H2O 0.35-0.49mg/L, H3BO38.50-11.4mg/L EDTA 30-50mg/L,
FeSO4·7H2O 2.50-4.98mg/L。
The present invention can make scenedesmus obliquus culture medium be beneficial to frustule fast-growth, the scenedesmus obliquus cell density turned out
Greatly, especially because with the addition of biological flocculant zymotic fluid.Nutrient formulation is reasonable, and formula components are simple, has saved certain culture
Cost；This culture medium can cultivate scenedesmus obliquus fast-growth under no light condition, i.e. heterotrophic growth can eliminate the limitation of light,
Expand the condition limitation of its application；Under this culture medium, economic microalgae scenedesmus obliquus secretes more autoflocculation material, is easy to wad a quilt with cotton
Retrogradation is formed sediment, and is advantageous to scenedesmus obliquus coagulation and recovery, reduces cost recovery；This culture medium may advantageously facilitate frustule fast breeding,
So that frustule concentration increases, culture systems stability is improved, is produced suitable for high-volume pilot scale culture, to produce in next step
Biodiesel improves biological raw material.
Brief description of the drawings
The shadow of Fig. 1 Different Nutritions concentration and its bioflocculant-producing bacteria composite flora concentration to scenedesmus obliquus fat content
Ring.
Embodiment
In order to which technical characteristic, purpose and the beneficial effect of the present invention is more clearly understood, 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 is described as follows with method below：
Various nutritive salt (in addition to glucose) used in the present invention can first be made into the higher mother liquor of concentration, generally
Mother liquor is configured to 1000 times of concentration, and when preparing mother liquor (1)-(7) number, medicine is prepared respectively, is deposited respectively；(8) number trace element
In medicine prepare according to quantity after mixed storage.After various mother liquors have been matched somebody with somebody, it is capped stores with Brown Glass Brown glass bottles and jars only respectively, it is labelled,
Indicate storage liquid title, prepare multiple, date etc., be stored in cold compartment of refrigerator.When preparing work liquid culture medium, first gently shake
Dynamic storage liquid, if it find that having precipitation, suspension or being polluted by Institute of Micro-biology, the storage liquid needs to match somebody with somebody again.Finally according to preparation
Culture volume with liquid-transfering gun from it is various storage liquid in take out required dosage respectively, needed during Ensure Liquid salt constantly plus
Water is simultaneously stirred, and various nutritive salt are added by the order provided in formula, be to avoid chemical reaction, are added distilled water diluting extremely
Required concentration.The culture medium prepared is adjusted to 7.0 with pH meter, packing, with 0.103MPa after wrapping, 121 DEG C, and autoclaving
20min。
Grease extracts and assay：Dry algae powder M1 is weighed, it is 1 to add certain volume ratio：2 chloroform：Methanol solution, shake
Mixing is swung, is extracted with ultrasonic disruption machine, time 20-30min；5min is centrifuged with 3000r/min again, takes chloroform layer molten
Liquid is repeated after extracting 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, is removed
Supernatant, frustule precipitation is washed with phosphate buffer twice, uses volume fraction to be resuspended for 20% dimethyl sulphoxide aqueous solution
Frustule, make algae solution OD5400.8,40 DEG C of water-bath 20min of value, adding 15 μ L Nile red dyes by 1m L algae solutions, (mass concentration is
0.1mg/m L 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 scenedesmus obliquus with mixed bacterial cogeneration system are identified
The water sample extracted from Harbin City biological flocculant manufacturing enterprise waste water treatment process aeration tank, load sterile take
Sample bag takes back laboratory, and microscope inspection observation finds that sample microbial and alga cells are more, to corresponding microalgae and microalgae companion
Raw flora is enriched with, separation screening.Raw water sample is launched in the reactor of simulation aeration biochemical pond and continuously cultivated, reactor is put
Under natural lighting, mixed with simulation bioflocculation agent production waste water well prepared in advance, long-time Air Exposure, oxygen content is protected
Hold in 4-5mg/L, 32 DEG C of cultures, pH 7.5, COD are maintained at 800mg/L or so, TN 90mg/L or so, TP 20mg/L or so.
Bioflocculation agent production waste water is simulated by the BG11 culture mediums of improvement, its simulated wastewater composition is following (mg/L)：Peptone
900, potassium dihydrogen phosphate 21.5, epsom salt 75.0, sodium acid carbonate 20, finally add micro- 1mL.It is wherein micro-
It is formulated as (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 500mg/L, addition 200mg/L glucose is as benefit
Fill carbon source.EGR is set in aeration tank, and 1.5L/min, equivalent 5h can be with circulation primary water samples so that water body flows back and mixes
It is even, continuous aeration pond culture more than 20d after, obtain one it is activity stabilized, the active bacterium algae that can be resistant to certain organic concentrations is total to
Raw system (as described below), number of viable maintains 107Individual/more than mL, microalgae quantity maintain 105Individual/mL.At the place of waste water
During reason, water sample starting COD be 200mg/L, the method that biochemical treatment use steps up waste water ratio, make functional flora by
Step adapts to high concentration simulation bioflocculation agent production waste water.The simulated wastewater of first generation domestication system addition 1/4, supplements 3/4
Distilled water, it is inoculated with flora, inoculum concentration 20% (v/v), 30 DEG C of shaking tables, 160rpm shaken cultivations.COD degradation rate is after 2d
50.56%, continue culture COD and keep stabilization no longer to decline.It is forwarded in second generation domestication culture medium, improves waste water ratio.
The simulated wastewater of 50% volume adds the distilled water of 50% volume, inoculum concentration 20%, 30 DEG C of shaking table cultures.After 2d detect COD from
Initial 412mg/L is reduced to 143.67mg/L, degradation rate 65.13%, and so on, into forth generation domestication system, trains
Support all simulation bioflocculation agent production waste waters of base, Biochemical Treatment such as table 1 institute of the active microalgae symbiosis flora to waste water
Show.COD is down to 131.36mg/L or so and keeps activity stabilized in sewage, and COD clearances are 84.00%, are given birth to raw sewage factory
It is close to change pond effluent quality, illustrates to have obtained one group of stable helotism system, has in terms of its COD in reduction system aobvious
Write effect.
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：Rhizobium (Rhizobium sp.), bacillus (Bacillus sp.), Klebsiella (Klebsiella
Sp.), enterobacteria (Enterobacter sp.), Aeromonas (Aeromonas sp.), Chryseobacterium sp
(Chryseobacterium sp.), sum to more than 80%.Wherein rhizobium (Rhizobium sp.) and bacillus
(Bacillus sp.) ratio highest, respectively reaches 20% and 18%, next to that Klebsiella (Klebsiella sp.),
Ratio reaches 15%, enterobacteria (Enterobacter sp.), Aeromonas (Aeromonas sp.) and Chryseobacterium sp
(Chryseobacterium sp.) accounts for 10%, 9% and 9% respectively.In activated bacterial microbiologic population, rhizobium
(Rhizobium sp.), bacillus (Bacillus sp.) and Klebsiella (Klebsiella sp.) are deposited extensively
It in nature and major sewage treatment plant's biochemistry pool, can be grown under the conditions of autotrophy or different oxygen, participate in carbon, nitrogen and phosphorus member
The materials such as element convert and circulation, can produce the active material with flocculation.Enterobacteria (Enterobacter) and gas
Zygosaccharomyces (Aeromonas sp.) has certain degradation of organic substances function, it may have the effect of denitrogenation dephosphorizing, is all kinds of sewage
In the microorganism that is widely present.Chryseobacterium sp (Chryseobacterium) is the function in municipal administration and food wastewater handling process
Flora, while can be with process part persistent organic pollutants.
By the above method and means, it is found that microalgae main species include two kinds of microalgaes：Grid algae (Scenedesmus sp.)
And chlorella (Chlorella sp.), wherein scenedesmus obliquus account for the 80% of sum.By Nile red decoration method to two plants of microalgaes
Fat content measure is carried out, it is found that grid algae fat content reaches 21.76%, total fat yield reaches 115.6mg/ (Ld), bead
Algae fat content reaches 17.25%, and total fat yield reaches 95.4mg/ (Ld).The present invention passes through 18S rDNA equimoleculars biology
Learn to do section to be identified, determine that the oil-rich microalgae grid algae (Scenedesmus sp.) in the inventive method is scenedesmus obliquus
(Scenedesmus obliquus)。
Embodiment 2：Compound flora and activity checking
Simulate bacterium for producing flocculant group rhizobium (Rhizobium sp.), bacillus in embodiment 1
(Bacillus sp.) and Klebsiella (Klebsiella sp.) proportion of composing, and scenedesmus obliquus (Scenedesmus
Obliquus) compounded.Wherein rhizobium (Rhizobium sp.) is from specific kind radiation rhizobium (Rhizobium
Radiobacter), bacillus (Bacillus sp.) is Bacillus sphaericus (Bacillus from specific kind
Sphaeicus), Klebsiella (Klebsiella sp.) is Klebsiella (Klebsiella from specific kind
variicola).Wherein single bacterium (algae) is activated with LB culture mediums respectively, and cell concentration is adjusted after activation, and scenedesmus obliquus addition is dense
Degree is more than three kinds of bacterium for producing flocculant concentration, two orders of magnitude, three kinds of bacterium for producing flocculant, i.e. rhizobium (Rhizobium
Sp.), bacillus (Bacillus sp.) and Klebsiella (Klebsiella sp.) are according to 4:4:3 are inoculated with.
Above bacterium algae is inoculated into embodiment 1 and simulated in bioflocculation agent production waste water, after cultivating 7 days, compounds obtained microorganism
Immobilization cell is according to fat content detection method described in embodiment, and analysis compounding flora is to scenedesmus obliquus fat content
Influence.Culture end retreads raw grid algae fat content for 25.58%, and grows stable, and algae cell density reaches 4.5 × 107Individual/
ML, total fat yield reach 138.7mg/ (Ld).The composite flora that the above results show to prepare demonstrates function bacterium in embodiment 1
Group is to the facilitation of scenedesmus obliquus Lipid-producing, while it is close to illustrate that composite flora that three kinds of single bacteriums compound to obtain grows for algae
The influence of degree, fat content and COD of sewage removal efficiency is better than the helotism system screened in embodiment 1, is respectively increased
900%, 17.56% and 7.14%.
Biological flocculant main component prepared by analysis shows is polysaccharide and a small amount of polypeptide, protein, lipid and its
Compound, by the mixed fermentation of three kinds of bacterium for producing flocculant, make polysaccharide of fermentation broth and protein classes more abundant, three kinds of bacterium it
Between there is symplastic growth effect, promote a large amount of secretions of metabolin each other, particular with active material-signal of specific function
Molecular species.Signaling molecule is a kind of exocytosis material of regulation biochemical function of microorganism secretion, when signaling molecule exists
Reach certain threshold value in culture environment, the expression of microorganism correlation biochemical function will be influenceed, promote growth metabolism.And above-mentioned life
Polysaccharide that thing flocculant contains, short chain polypeptides class material and the AHLs classes material (signaling molecule) that contains are that microalgae is quick
Growth and the promotive factor of oil and fat accumulation, will play value-added effect to micro algae growth.
Embodiment 3：Different Nutrition concentration and biological flocculant addition concentration to scenedesmus obliquus grow and fat content it is excellent
Change
The scenedesmus obliquus being formulated as follows with sterilized water configuration optimizes wild Oryza species：Glucose 10000mg/L, NaNO3
150mg/L, MgSO4·7H2O 50mg/L, K2HPO450mg/L, MnCl2·4H2O 1.00mg/L, Co (NO3)2·6H2O
0.35mg/L, H3BO38.50mg/L, EDTA 30mg/L, FeSO4·7H2O 2.50mg/L.Addition includes rhizobium
(Rhizobium sp.) radiation rhizobium (Rhizobium radiobacter), bacillus (Bacillus sp.) ball
Shape bacillus (Bacillus sphaeicus) and Klebsiella (Klebsiella sp.) Klebsiella
The mixed bacterial zymotic fluid 10mL of (Klebsiella variicola), wherein three kinds of strain inoculation proportionings are 4:4:3.It is mixed
Flora biological flocculant zymotic fluid concrete composition and fermentation condition are as follows：Glucose 10g/ml, K2HPO45g/L, MgSO4·
7H2O 0.2g/L, urea 0.5g/L, H2PO42g/L, NaCl 0.1g/L, yeast extract 0.5g/L, pH 7.5.Three kinds of flocculants
Producing strains press 4：4：3 ratios are inoculated into seed culture fluid, expand culture bacterium solution used for before by required fermentation liquid measure
10% seed culture fluid prepared, shaking table 140r/min, as 30 DEG C, fermentation time 24h, biological flocculant zymotic fluid.Take pair
The scenedesmus obliquus in number growth period, which is inoculated into the culture medium, to be cultivated, and 24 hours continuous lights, intensity is about 8000Lux, training
The degrees centigrade of temperature control 30 is supported, shakes twice manually daily, prevents from precipitating adherent, scenedesmus obliquus in periodic logging culture medium
Frustule concentration.BG-11 culture mediums are used to make as control medium, not additional mixed bacterial biological flocculant zymotic fluid in addition
For control experiment group.Each experimental group algae density value is measured in the exponential phase middle and later periods, refers to table 2；Simultaneously to scenedesmus obliquus oil
Fat content is measured, and refers to Fig. 1.
The Different Nutrition concentration of table 2 and biological flocculant add concentration to scenedesmus obliquus growth effect contrast table
Embodiment 4：Different Nutrition concentration and biological flocculant addition concentration to scenedesmus obliquus grow and fat content it is excellent
Change
The scenedesmus obliquus being formulated as follows with sterilized water configuration optimizes wild Oryza species：Glucose 150000mg/L, NaNO3
250mg/L, MgSO4·7H2O 75mg/L, K2HPO4175mg/L, MnCl2·4H2O 1.44mg/L, Co (NO3)2·6H2O
0.49mg/L, H3BO311.4mg/L, EDTA 50mg/L, FeSO4·7H2O 4.98mg/L.Add mixed bacterial bioflocculation
Agent zymotic fluid 10mL, mixed bacterial zymotic fluid group is into, fermentation condition and bacterium algae inoculative proportion with embodiment 3.Take the logarithm growth period
Scenedesmus obliquus be inoculated into the culture medium and cultivated, 24 hours continuous lights, intensity is about 8000Lux, cultivation temperature control
30 degrees centigrades are made, shakes twice manually daily, prevents from precipitating adherent, the frustule of scenedesmus obliquus in periodic logging culture medium
Concentration.Using BG-11 culture mediums in addition, not additional mixed bacterial biological flocculant zymotic fluid is as control as control medium
Experimental group.Each experimental group algae density value is measured in the exponential phase middle and later periods, refers to table 3；Simultaneously to scenedesmus obliquus fat content
It is measured, refers to Fig. 1.
The Different Nutrition concentration of table 3 and biological flocculant add concentration to scenedesmus obliquus growth effect contrast table
Embodiment 5：Different Nutrition concentration and biological flocculant addition concentration to scenedesmus obliquus grow and fat content it is excellent
Change
The scenedesmus obliquus being formulated as follows with sterilized water configuration optimizes wild Oryza species：Glucose 10000mg/L, NaNO3
150mg/L, MgSO4·7H2O 50mg/L, K2HPO450mg/L, MnCl2·4H2O 1.00mg/L, Co (NO3)2·6H2O
0.35mg/L, H3BO38.50mg/L, EDTA 30mg/L, FeSO4·7H2O 2.50mg/L.Add mixed bacterial bioflocculation
Agent zymotic fluid 20mL, mixed bacterial zymotic fluid group is into, fermentation condition and bacterium algae inoculative proportion with embodiment 3.Take the logarithm growth period
Scenedesmus obliquus be inoculated into the culture medium and cultivated, 24 hours continuous lights, intensity is about 8000Lux, cultivation temperature control
30 degrees centigrades are made, shakes twice manually daily, prevents from precipitating adherent, the frustule of scenedesmus obliquus in periodic logging culture medium
Concentration.Using BG-11 culture mediums in addition, not additional mixed bacterial biological flocculant zymotic fluid is as control as control medium
Experimental group.Each experimental group algae density value is measured in the exponential phase middle and later periods, refers to table 4；Simultaneously to scenedesmus obliquus fat content
It is measured, refers to Fig. 1.
The Different Nutrition concentration of table 4 and biological flocculant add concentration to scenedesmus obliquus growth effect contrast table
Embodiment 6：Different Nutrition concentration and biological flocculant addition concentration to scenedesmus obliquus grow and fat content it is excellent
Change
The scenedesmus obliquus being formulated as follows with sterilized water configuration optimizes wild Oryza species：Glucose 150000mg/L, NaNO3
250mg/L, MgSO4·7H2O 75mg/L, K2HPO4175mg/L, MnCl2·4H2O 1.44mg/L, Co (NO3)2·6H2O
0.49mg/L, H3BO311.4mg/L, EDTA 50mg/L, FeSO4·7H2O 4.98mg/L.Add mixed bacterial bioflocculation
Agent zymotic fluid 20mL, mixed bacterial zymotic fluid group is into, fermentation condition and bacterium algae inoculative proportion with embodiment 3.Take the logarithm growth period
Scenedesmus obliquus be inoculated into the culture medium and cultivated, 24 hours continuous lights, intensity is about 8000Lux, cultivation temperature control
30 degrees centigrades are made, shakes twice manually daily, prevents from precipitating adherent, the frustule of scenedesmus obliquus in periodic logging culture medium
Concentration.Using BG-11 culture mediums in addition, not additional mixed bacterial biological flocculant zymotic fluid is as control as control medium
Experimental group.Each experimental group algae density value is measured in the exponential phase middle and later periods, refers to table 5；Simultaneously to scenedesmus obliquus fat content
It is measured, refers to Fig. 1.
The Different Nutrition concentration of table 5 and biological flocculant add concentration to scenedesmus obliquus 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 can make scenedesmus obliquus fast breeding, scenedesmus obliquus is entered increased logarithmic phase in advance, shorten its incubation time.Can from Fig. 1
To find out, each oblique raw grid algae oil fat content balance control medium BG-11 of embodiment optimal culture conditions has carrying for conspicuousness
Height, thus illustrate that scenedesmus obliquus cultural method provided by the invention has obvious culture advantage, more conducively frustule is quick
Growth, while promote fat content to improve.
The schematical embodiment of the present invention is the foregoing is only, is not limited to the scope of the present invention.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 scenedesmus obliquus, it is characterised in that described cultural method is the culture in culture scenedesmus obliquus
Compound biological flocculant zymotic fluid is added in base, described zymotic fluid is by Klebsiella (Klebsiella sp.)；
Prepared by rhizobium (Rhizobium sp.) and bacillus (Bacillus sp.) fermentation.
2. the method as described in claim 1, it is characterised in that described Klebsiella (Klebsiella sp.) is gram
The primary Salmonella of thunder (Klebsiella variicola).
3. the method as described in claim 1, it is characterised in that described rhizobium (Rhizobium sp.) bacterium is to put
Penetrate shape rhizobium (Rhizobium radiobacter).
4. the method as described in claim 1, it is characterised in that described bacillus (Bacillus sp.) bacterium is ball
Shape bacillus (Bacillus sphaeicus).
5. the method as described in claim 1, it is characterised in that described compound biological flocculant zymotic fluid, what it was used
The composition of fermentation medium is as follows：Glucose 10g/ml, K2HPO45g/L, MgSO4·7H2O 0.2g/L, urea 0.5g/L,
H2PO42g/L, NaCl 0.1g/L, yeast extract 0.5g/L, pH 7.5.
6. the method as described in claim 1, it is characterised in that the culture medium of described culture scenedesmus obliquus, its formula are as follows：
Glucose 10000-150000mg/L, NaNO3150-250mg/L, MgSO4·7H2O 50-75mg/L, K2HPO4 50-175mg/
L, MnCl2·4H2O 1.00-1.44mg/L, Co (NO3)2·6H2O 0.35-0.49mg/L, H3BO38.50-11.4mg/L
EDTA 30-50mg/L, FeSO4·7H2O 2.50-4.98mg/L。
7. a kind of culture medium of scenedesmus obliquus, it is characterised in that sent out in described culture medium added with compound biological flocculant
Zymotic fluid, described zymotic fluid are by Klebsiella (Klebsiella sp.)；Rhizobium (Rhizobium sp.) and bud
It is prepared by spore Bacillus (Bacillus sp.) fermentation.
8. culture medium as claimed in claim 7, it is characterised in that the primary Bordetella of described thunder (Klebsiella sp.) bacterium
For Klebsiella (Klebsiella variicola).
9. culture medium as claimed in claim 7, it is characterised in that described rhizobium (Rhizobium sp.) bacterium is
Radiation rhizobium (Rhizobium radiobacter).
10. culture medium as claimed in claim 7, it is characterised in that described bacillus (Bacillus sp.) bacterium
For Bacillus sphaericus (Bacillus sphaeicus).
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| CN109504716A (en) * | 2019-01-29 | 2019-03-22 | 西北农林科技大学 | A kind of cultural method promoting scenedesmus obliquus oil-producing |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108220165A (en) * | 2017-12-26 | 2018-06-29 | 仲恺农业工程学院 | A kind of method for strengthening microalgae removal breeding wastewater carbon nitrogen and phosphorus pollution using association flora |
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Application publication date: 20171208 |