CN102093955A - Chlorella strain and application thereof - Google Patents
Chlorella strain and application thereof Download PDFInfo
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- CN102093955A CN102093955A CN2010105725254A CN201010572525A CN102093955A CN 102093955 A CN102093955 A CN 102093955A CN 2010105725254 A CN2010105725254 A CN 2010105725254A CN 201010572525 A CN201010572525 A CN 201010572525A CN 102093955 A CN102093955 A CN 102093955A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Abstract
The invention discloses a Chlorella strain and an application thereof. The Chlorella strain provided by the invention is Chlorella sp. YL-2 of which collection number is CGMCC No.4305. The Chlorella sp. YL-2 can well grow in sewage, and can efficiently remove nitrogen and phosphorus in sewage. The Chlorella sp. YL-2 is rich in oil and fat, and can be used for preparing biodiesel. Therefore, the Chlorella sp. YL-2 has wide application prospects in culture using sewage and biodiesel conversion system.
Description
Technical field
The present invention relates to a chlorella and application thereof, be specifically related to the application of this chlorella in bioenergy and sewage purification Application Areas.
Background technology
Energy dilemma and water resources crisis are the current two big crises that face in the world.According to relevant, global mineral fossil energy reserve: 1,500 hundred million tons in oil, but be 41 years working life; 5,500 hundred million tons of coals, but be 200 years working life; 1,100 hundred million tons of Sweet natural gases, but be 60 years working life.The socioeconomic continuous increase that develops rapidly with population, limited Freshwater resources have been subjected to severe contamination and destruction, and the whole world is being faced with the severe crisis of water scarcity.Therefore, reproducible bioenergy of development of new (biofuel) and saprobia advanced treatment new technology are to tackle the serious day by day energy dilemma and the effective measure of water resources crisis, also are the only ways of human kind sustainable development.
Utilizing little algae to prepare biofuel has become current domestic and international research focus, especially utilizes sewage to cultivate little algae, and preparing biofuel when purifying waste water is the advanced subject of this research field.Little algae has that growth is fast, harvesting time short, be rich in characteristics (annual fixed CO such as grease, photosynthetic utilising efficiency height
2Account for greatly global net photosynthesis output 40%), be the main raw material(s) of present alternative fossil energy.Therefore can absorb a large amount of nitrogen phosphoric in the micro algae growth process, can utilize the post-processing unit of sewage work to cultivate little algae and carry out denitrogenation dephosphorizing deep purifying sewage.
Can at present, the domestic development be cultivated little algae about sewage to prepare the correlation technique research report of biofuel less, obtain that suitable sewage is cultivated and to be rich in grease higher, and the little algae algae strain that is easy to transform the preparation biofuel is crucial.From existing report, the algae strain ubiquity culture condition that is utilized requires problems such as height, biological yield and grease yield are low, biofuel transforms instability to be difficult to realize using.Therefore, the good algae kind of screening has great importance for addressing the above problem.
Summary of the invention
The purpose of this invention is to provide a chlorella and application thereof.
Chlorella provided by the invention (Chlorella sp.), called after YL-2, separation is from the water sample in Qinghe, Beijing, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on November 09th, 2010 and (be called for short CGMCC, the address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City), culture presevation number is CGMCC No.4305.Chlorella YL-2 all can grow under the scope of the culture temperature of 4 ℃-37 ℃ (preferred 10 ℃-35 ℃, most preferably 28 ℃) and pH6-9 (preferred pH7).
Chlorella (Chlorella sp.) YL-2CGMCC No.4305 is called for short chlorella YL-2.
The present invention also protects a kind of seed culture medium that is used to cultivate chlorella (Chlorella sp.) YL-2, following solute water is settled to 1 liter obtains: 1ml A
5+ Co solution, 2g starch, 1.5g peptone, 1.5g NaNO
3, 0.04gK
2HPO
43H
2O, 0.075gMgSO
47H
2O, 0.036gCaCl
22H
2O, 0.006g citric acid, 0.006g ferric citrate amine, 0.001g Na
2EDTA and 0.02g Na
2CO
3Described A
5+ Co solution is settled to 1 liter with following solute water and obtains: 2.86g/L H
3BO
3, 1.81g/L MnCl
2H
2O, 0.222g/L ZnSO
47H
2O, 0.079g/LCuSO
45H
2O, 0.390g/L Na
2MoO
42H
2O, 0.049g/L Co (NO
3)
26H
2O.
The present invention also protects a kind of method for preparing chlorella (Chlorella sp.) YL-2 seed liquor, is to cultivate chlorella (Chlorella sp.) YL-2CGMCC No.4305 in described seed culture medium, obtains seed liquor.
The temperature condition of cultivating chlorella (Chlorella sp.) YL-2CGMCC No.4305 specifically can be 4 ℃-37 ℃, and preferred 10 ℃-35 ℃, most preferably 28 ℃.The pH condition of cultivating chlorella (Chlorella sp.) YL-2CGMCC No.4305 specifically can be pH4-9, preferred pH6-9, most preferably pH7.
Chlorella (Chlorella sp.) YL-2CGMCC No.4305 can be used for sewage disposal.Described sewage disposal can be specifically that sewage is denitrogenated and/or sewage dephosphorization.
Chlorella (Chlorella sp.) YL-2CGMCC No.4305 can be used for preparing biofuel (fatty acid methyl ester).
The present invention also protects a kind of preparation method of bio-diesel oil, be with the grease that extracts among chlorella (Chlorella sp.) YL-2CGMCCNo.4305 as raw material, the preparation biofuel.Chlorella (Chlorella sp.) YL-2CGMCCNo.4305 can cultivate in sewage earlier before carrying out described extraction.Specifically described seed liquor can be inoculated in the described sewage and be cultivated.
Chlorella YL-2 well-grown in sewage can efficiently be removed the nitrogen phosphorus in the sewage simultaneously.Chlorella YL-2 self is rich in the grease height, can be used to prepare biofuel.In sum, chlorella YL-2 has broad application prospects in utilizing sewage cultivation and biofuel transformation system.
Description of drawings
Fig. 1 is the phylogenetic tree that makes up based on 18S rDNA partial sequence.
Fig. 2 is the growing state of chlorella YL-2 on different basic mediums.
Fig. 3 is for adding the influence of various carbon sources to chlorella YL-2 growth.
Fig. 4 is for adding the influence of various nitrogenous sources to chlorella YL-2 growth.
Fig. 5 is the influences of various pH conditions to chlorella YL-2 growth.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment if no special instructions, is ordinary method.Used test materials among the following embodiment if no special instructions, is to buy from routine biochemistry reagent shop and obtains.Quantitative test in following examples all is provided with repeated experiments three times, results averaged.
The isolation identification of embodiment 1, chlorella YL-2
One, sample collecting
Gather the water sample that contains green alga in July, 2010 from Qinghe, Beijing.
Two, the separation of chlorella YL-2 and screening
Separating step: the water sample of gathering is transferred in the sanitary sewage (taking from certain city domestic sewage treatment plant) through autoclaving (121 ℃, high pressure steam sterilization 20 minutes), place the illumination box enrichment culture.Carry out after the three-wheel enrichment culture, suitably dilute illumination cultivation behind algae liquid and the sewage agar semisolid medium mixing, choose single algae and fall within enlarged culturing in the small test tube.
Screening step: at first pure algae strain is inoculated in 48 hole enzyme plates (aseptic sewage 0.25mL is contained in every hole), place in the illumination box and cultivate, detect growing state with 540nm, select the algae strain faster of growing, and measure its fat content under the sewage culture condition.
The parameter of liquid culture in sewage: 28 ± 1 ℃; 1000～1200lux intensity of illumination; Light To Dark Ratio 14h: 10h; Regularly vibrate every day (2 times), avoid cell precipitation; Cultivated 7 days.
Obtain a strain and in sewage, grow comparatively fast, be rich in greasy pure algae strain, called after YL-2.
Three, the evaluation of chlorella YL-2
1, the identification of morphology of chlorella YL-2
Rounded or the elliposoidal of cellular form, cell dia 3-6 μ m has a tangible chromatophore, light green in the middle of the Individual existence, cell.
2, the Molecular Identification of chlorella YL-2
Collecting the cell of exponential phase of growth, extract test kit (the prosperous Bioisystech Co., Ltd of Beijing ancient cooking vessel state) with plant genome DNA and extract genomic dna, is template amplification 18S rDNA gene fragment with it.The pcr amplification system is 25 μ L, wherein contains 12.5 μ L PCR 2 * mix (TianGen), forward and reverse primer (10 μ mol/L) each 1 μ L, template DNA 1 μ L, two steaming aqua sterilisa 9.5 μ L.The pcr amplification program is: 95 ℃ of pre-sex change 3min; 94 ℃ of sex change 1min, 55 ℃ of renaturation 1min, 72 ℃ are extended 1.5min, totally 30 circulations; 72 ℃ are extended 10min.
The primer of 18S rDNA of being used to increase is a universal primer, and sequence is as follows:
Forward primer (18SF): 5 '-CCTGGTTGATCCTGCCAG-3 ';
Reverse primer (18SR): 5 '-TTGATCCTTCTGCAGGTTCA-3 '.
Pcr amplification product reclaims test kit with glue and reclaims after 1% agarose gel electrophoresis separates, transfer to Shanghai and give birth to the order-checking of worker Bioisystech Co., Ltd.Pcr amplification product is a 18s rDNA partial sequence, is total to 1742bp, and sequencing result is shown in the sequence 1 of sequence table.
Sequencing result is through NCBI website BLAST compare of analysis, and the Query coverage value of YL-2 and Chlorella vulgaris (FM205855) reaches 100%, and Max ident is 99.9%.Carry out the multisequencing homology analysis with MEGA 4.0 softwares, and constructing system is grown tree (see Fig. 1, the numerical value on the branch is the result of bootstrapping 1000 times).Algae YL-2 and Chlorella vulgaris gather well in a branch.
Based on the result of identification of morphology and Molecular Identification, YL-2 is accredited as chlorella (Chlorella sp.).YL-2 is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center with chlorella (Chlorella sp.), and preserving number is No.4305.
Determining of embodiment 2, chlorella YL-2 seed culture medium
One, the selection of basic medium
Utilize the sewage large-scale culturing micro-algae to need a certain amount of elite seed, therefore investigate and cultivate of the influence of the common substratum (BG11 substratum, SE substratum, aquatic No. 6 substratum and f/2 substratum) that adopts of chlorella algae strain YL-2 growth.
1, the preparation of substratum
(1) BG11 substratum
Preparation BG11 substratum (pH7.0).
BG11 substratum (pH7.0) is by NaNO
3, K
2HPO
43H
2O, MgSO
47H
2O, CaCl
22H
2O, citric acid (citricacid), ferric citrate amine (Ferric ammonium citrate), Na
2EDTA, Na
2CO
3, A
5+ Co solution and distilled water are formed; In every liter of substratum, contain 1ml A
5+ Co solution, 1.5g NaNO
3, 0.04g K
2HPO
43H
2O, 0.075g MgSO
47H
2O, 0.036g CaCl
22H
2O, 0.006g citric acid, 0.006g ferric citrate amine, 0.001gNa
2EDTA and 0.02g Na
2CO
3
A
5+ Co solution is made up of solute and water; Solute and concentration thereof are: 2.86g/L H
3BO
3, 1.81g/L MnCl
2H
2O, 0.222g/L ZnSO
47H
2O, 0.079g/L CuSO
45H
2O, 0.390g/L Na
2MoO
42H
2O, 0.049g/LCo (NO
3)
26H
2O.
(2) SE substratum
Preparation SE substratum (pH7.0).
SE substratum (pH7.0) is by NaNO
3, K
2HPO
43H
2O, MgSO
47H
2O, CaCl
22H
2O, KH
2PO
4, NaCl, FeCl
36H
2O, Fe-EDTA, soil extract, A
5Solution and distilled water are formed; In every liter of SE substratum, contain 1ml Fe-EDTA, 40ml soil extract, 1ml A
5Solution, 0.25g NaNO
3, 0.075g K
2HPO
43H
2O, 0.075g MgSO
47H
2O, 0.025g CaCl
22H
2O, 0.175g KH
2PO
4, 0.025g NaCl, 0.005gFeCl
36H
2O.
The preparation method of soil extract: get 0.5kg and do not execute overplumped garden soil, place triangular flask (or beaker), add 1000ml distilled water, seal bottleneck with porous plug, the boiling water heating is 2 hours in water-bath, cooling number hour filters under aseptic condition, gets supernatant liquor and adds sterile purified water to cumulative volume 1000ml; 4 ℃ of preservations are standby.
A
5Solution is made up of solute and water; Solute and concentration thereof are: H
3BO
32.86g/L, MnCl
24H
2O 1.81g/L, ZnSO
47H
2O 0.22g/L, CuSO
45H
2O 0.009g/L, (NH
4)
6Mo
7O
244H
2O 0.004g/L.
The preparation method of Fe-EDTA: with lg Na
2EDTA is dissolved in 50ml distilled water; With 8lmg FeCl
36H
2O is dissolved in the HCl aqueous solution of 50ml 0.1M; Above two solution are mixed.
(3) aquatic No. 6 substratum
Prepare aquatic No. 6 substratum (pH7.0).
SE substratum (pH7.0) is by NH
2CONH
2, MgSO
47H
2O, NaHCO
3, KCl, FeSO
4, CaCl
2, soil extract and distilled water forms; In every liter of substratum, contain 0.5ml soil extract, 0.133g/L NH
2CONH
2, 0.1g/LMgSO
47H
2O, 0.1g/L NaHCO
3, 0.033g/L KCl, 0.2g/L FeSO
4, 0.03g/L CaCl
2
The preparation method of soil extract is with step 2.
(4) f/2 substratum
Preparation f/2 substratum (pH7.0).
F/2 substratum (pH7.0) is by NaHCO
3, NaCl, KH
2PO
4, KNO
3, NH
2CONH
2, MgSO
47H
2O, f/2 trace element solution, f/2 vitamin solution and distilled water are formed; In every liter of substratum, contain 1ml f/2 trace element solution, 1ml f/2 vitamin solution, 12g NaHCO
3, 1g NaCl, 0.02g KH
2PO
4, 0.1g KNO
3, 0.03gNH
2CONH
2, 0.02g MgSO
47H
2O.
The preparation method of f/2 trace element solution: with ZnSO
44H
2O 0.023g, MnCl
24H
2O 0.078g, CuSO
45H
2O 0.010g, FeC
6H
5O
75H
2O 0.0039g, Na
2MoO
42H
2O 0.0073g, CoCl
26H
2O0.012g, Na
2EDTA 0.00435g is dissolved in deionized water, and is settled to 1000ml with deionized water.
The preparation method of f/2 vitamin solution: with vitamin B12 0.0005g, vitaminB10 .1g is dissolved in deionized water, and is settled to 1000ml with deionized water.
2, the selection of basic medium
Chlorella YL-2 is seeded to BG11 substratum, SE substratum, aquatic No. 6 substratum or f/2 substratum respectively (with OD
540nmTo be 0.5 chlorella YL-2 be inoculated in each substratum the OD of chlorella YL-2 in substratum with 20% volume ratio to value
540nmValue is about 0.15), cultivated 8 days for 28 ℃.
Detect the OD of each nutrient solution
540nmValue the results are shown in Figure 2.Chlorella YL-2 growth differences in four kinds of basic mediums is bigger, and it is the fastest to grow when adopting the BG11 culture medium culturing, has obtained bigger biomass (OD when cultivating 8 days
540nmReach 1.13), and growth is the poorest in the f/2 substratum.Therefore, the BG11 substratum is comparatively suitable basic medium.
Two, chlorella YL-2 seed culture medium determines
For making the faster and better growth of chlorella YL-2, on the BG11 medium base, carry out the optimization of nitrogenous source, carbon source.
Add different carbon sources on the BG11 medium base, the concentration of each carbon source in substratum is 2g.Chlorella YL-2 is seeded to each substratum respectively (with OD
540nmTo be 0.5 chlorella YL-2 be inoculated in each substratum the OD of chlorella YL-2 in substratum with 20% volume ratio to value
540nmValue is about 0.15), cultivated 8 days for 28 ℃.Detect the OD of each nutrient solution
540nmValue the results are shown in Figure 3.
Add different nitrogen sources on the BG11 medium base, the concentration of each nitrogenous source in substratum is 1.5g.Chlorella YL-2 is seeded to each substratum respectively (with OD
540nmTo be 0.5 chlorella YL-2 be inoculated in each substratum the OD of chlorella YL-2 in substratum with 20% volume ratio to value
540nmValue is about 0.15), cultivated 8 days for 28 ℃.Detect the OD of each nutrient solution
540nmValue the results are shown in Figure 4.
The BG11 substratum is adjusted to different initial pH, chlorella YL-2 is seeded to each substratum respectively (with OD
540nmTo be 0.5 chlorella YL-2 be inoculated in each substratum the OD of chlorella YL-2 in substratum with 20% volume ratio to value
540nmValue is about 0.15), cultivated 8 days for 28 ℃.Detect the OD of each nutrient solution
540nmValue the results are shown in Figure 5.
The result shows that optimum carbon source is a starch, and optimum nitrogen source is a peptone, and best pH is 7.0 (all can grow under the scope of pH6-9).
According to the optimum result of nitrogenous source, carbon source, determine the prescription of seed culture medium, preparation seed culture medium (pH7.0).
Seed culture medium is settled to 1 liter with following solute water and obtains: add 1ml A
5+ Co solution, 2g starch, 1.5g peptone, 1.5g NaNO
3, 0.04g K
2HPO
43H
2O, 0.075g MgSO
47H
2O, 0.036g CaCl
22H
2O, 0.006g citric acid, 0.006g ferric citrate amine, 0.001g Na
2EDTA and 0.02g Na
2CO
3
A
5+ Co solution is settled to 1 liter with following solute water and obtains: 2.86g/L H
3BO
3, 1.81g/LMnCl
2H
2O, 0.222g/L ZnSO
47H
2O, 0.079g/L CuSO
45H
2O, 0.390g/L Na
2MoO
42H
2O, 0.049g/L Co (NO
3)
26H
2O.
The growing state of embodiment 3, chlorella YL-2
Raw waste water is taken from certain city domestic sewage treatment plant, and through precipitation, unsterilised, condition of water quality sees Table 1.
Table 1 raw waste water water quality situation
| Water-quality guideline | Desired value |
| Total nitrogen (mg/L) | 2091.63 |
| Ammonia nitrogen (mg/L) | 349.09 |
| Total phosphorus (mg/L) | 84.2 |
| CODcr(mg/L) | 3502 |
| pH | 6.9 |
1, the cultivation of chlorella YL-2
In the seed culture medium of embodiment 2 preparations, cultivate chlorella YL-2 to logarithmic phase (OD for 28 ℃
540nmBe about 0.8), be forwarded to (the bottled 500ml raw waste water of each 1000ml triangle) in the triangular flask with 20% (volume ratio), 28 ℃, intensity of illumination 1000～1200lux, Light To Dark Ratio 14h: cultivate 7 days (growing into lag phase) under the 10h condition.
2, biomass and fat content are measured
Get nutrient solution and place centrifuge tube, the centrifugal 3min of 5000r/min, abandoning supernatant with the uncovered baking oven that places of centrifuge tube, is dried to constant weight for 80 ℃.Every liter of nutrient solution obtains the dry frond (biomass is 0.57g/L) of 0.57g.
Dry frond ground fully (weight is W for the algae powder
0), add solvent (sherwood oil: ether=2: 1; Volume ratio), 50 ℃ of lixiviates 5 hours, during suitable vibration mixing; After extracting end, add 10% (quality percentage composition) potassium hydroxide aqueous solution sedimentation cell, shake up the centrifugal 10min of static back 6000r/min, collect supernatant liquor in the centrifuge tube of drying and constant weight, boil off unnecessary solvent rapidly in 60 ℃ of water-baths, (weight is W to obtain the chlorella grease
1), the fat content (W of calculating chlorella
1/ W
0* 100%).The result shows that chlorella YL-2 cultivates the back fat content in raw waste water higher, is about 39.2%.
Certain city domestic sewage treatment plant is taken from second-level settling pond water outlet (B-grade sewage), and unsterilised, condition of water quality sees Table 2.
Table 2 B-grade sewage water quality situation
| Water-quality guideline | Desired value |
| Total nitrogen (mg/L) | 2326.21 |
| Ammonia nitrogen (mg/L) | 1.73 |
| Total phosphorus (mg/L) | 1.17 |
| CODcr(mg/L) | 25.3 |
| Suspended substance (mg/L) | 9.6 |
| Turbidity | 1.85 |
| pH | 7.02 |
1, the cultivation of chlorella YL-2
In the seed culture medium of embodiment 2 preparations, cultivate chlorella YL-2 to logarithmic phase (OD for 28 ℃
540nmBe about 0.8), be forwarded to (the bottled 500ml B-grade sewage of each 1000ml triangle) in the triangular flask with 20% (volume ratio), 28 ℃, intensity of illumination 1000～1200lux, Light To Dark Ratio 14h: cultivate 8 days (growing into stationary phase) under the 10h condition, regularly vibrate every day (2 times), avoid cell precipitation.
2, the mensuration of removal efficiency of nitrogen and phosphorus
Get nutrient solution and carry out determination of total nitrogen content (GB11894-89, the mensuration of water quality total nitrogen alkalescence alkaline potassium per-sulfate digestion ultraviolet spectrophotometry) and total phosphorus determination (GB/T 11893-1989, the mensuration ammonium molybdate spectrophotometry of water quality total phosphorus), the results are shown in Table 3 (3 bottles mean values).
Total nitrogen and total phosphorous behind the table 3 cultivation chlorella YL-2 in the B-grade sewage
3, biomass and fat content are measured
Measuring method is with the step 2 of embodiment 3
Every liter of nutrient solution obtains 0.45g exsiccant frond (biomass is 0.45g/L).
The fat content of chlorella is 42.2%.
The result shows, chlorella YL-2 fat content is higher, and well-grown in B-grade sewage has stronger removal effect to nitrogen (phosphorus source deficiency cause nitrogen removal rate lower), phosphorus, is a strain very potential little algae in the production application of sewage disposal coupled biological oil.
1, the cultivation of chlorella YL-2
2, preparation chlorella grease
Get nutrient solution and place centrifuge tube, the centrifugal 3min of 5000r/min, abandoning supernatant with the uncovered baking oven that places of centrifuge tube, is dried to constant weight for 80 ℃.
Dry frond fully ground be the algae powder, add solvent (sherwood oil: ether=2: 1; Volume ratio), 50 ℃ of lixiviates 5 hours, during suitable vibration mixing; After extracting end, add 10% (quality percentage composition) potassium hydroxide sedimentation cell, shake up the centrifugal 10min of static back 6000r/min, collect supernatant liquor in the centrifuge tube of drying and constant weight, in 60 ℃ of water-baths, boil off unnecessary solvent rapidly, obtain the chlorella grease.
3, preparation biofuel
Chlorella grease and methyl alcohol are carried out transesterification, obtain fatty acid methyl ester, i.e. biofuel.Processing condition: alcohol oil rate 30: 1 (amount of substance ratio); Catalyzer is KOH, and consumption is 1% of a chlorella oil quality, and temperature of reaction is 50 ℃, reaction times 1h.Reaction finishes the back centrifugation, obtains fatty acid methyl ester behind the upper strata underpressure distillation excessive methanol, and lower floor is a byproduct glycerine.
Repeated experiments is set three times, results averaged.The greasy quality * 100% of the quality/chlorella of the yield=fatty acid methyl ester of fatty acid methyl ester (biofuel).The yield of fatty acid methyl ester (biofuel) is 69.6%.
Claims (10)
1. chlorella (Chlorella sp.) YL-2, its deposit number is CGMCC No.4305.
2. be used to cultivate the seed culture medium of chlorella (Chlorella sp.) YL-2, following solute water be settled to 1 liter obtain: 1ml A
5+ Co solution, 2g starch, 1.5g peptone, 1.5g NaNO
3, K O.04g
2HPO
43H
2O, 0.075g MgSO
47H
2O, 0.036g CaCl
22H
2O, 0.006g citric acid, 0.006g ferric citrate amine, 0.001gNa
2EDTA and 0.02g Na
2CO
3
Described A
5+ Co solution is settled to 1 liter with following solute water and obtains: 2.86g/L H
3BO
3, 1.81g/LMnCl
2H
2O, 0.222g/L ZnSO
47H
2O, 0.079g/L CuSO
45H
2O, 0.390g/L Na
2MoO
42H
2O, 0.049g/L Co (NO
3)
26H
2O.
3. a method for preparing chlorella (Chlorella sp.) YL-2 seed liquor is to cultivate chlorella (Chlorella sp.) YL-2CGMCC No.4305 in the described seed culture medium of claim 2, obtains seed liquor.
4. method according to claim 3 is characterized in that: the temperature condition of described cultivation chlorella (Chlorella sp.) YL-2CGMCC No.4305 is 4 ℃-37 ℃, preferred 10 ℃-35 ℃, and most preferably 28 ℃; The pH condition of described cultivation chlorella (Chlorella sp) YL-2CGMCC No.4305 is pH4-9, preferred pH6-9, most preferably pH7.
5. chlorella (Chlorella sp) YL-2CGMCC No.4305 is in Application of Sewage.
6. method as claimed in claim 6 is characterized in that: described sewage disposal is that sewage is denitrogenated and/or sewage dephosphorization.
7. the application of chlorella (Chlorella sp) YL-2CGMCC No.4305 in the preparation biofuel.
8. one kind prepares method of bio-diesel oil, be with the grease that extracts among chlorella (Chlorella sp.) the YL-2CGMCC No.4305 as raw material, the preparation biofuel.
9. method as claimed in claim 8 is characterized in that: chlorella (Chlorella sp.) YL-2CGMCCNo.4305 cultivates in sewage earlier before carrying out described extraction.
10. method as claimed in claim 9 is characterized in that: described cultivation is cultivated for claim 3 or 4 described seed liquor are inoculated in the described sewage.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105725254A CN102093955B (en) | 2010-12-03 | 2010-12-03 | Chlorella strain and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105725254A CN102093955B (en) | 2010-12-03 | 2010-12-03 | Chlorella strain and application thereof |
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| CN102746992A (en) * | 2012-07-16 | 2012-10-24 | 哈尔滨工业大学 | Method for culturing chlorella by heterotrophism with sludge hydrolysate |
| WO2013013433A1 (en) * | 2011-07-22 | 2013-01-31 | 中国科学院烟台海岸带研究所 | Chlorella mutant strain and application thereof |
| CN103468578A (en) * | 2013-10-14 | 2013-12-25 | 哈尔滨工业大学 | Chlorella strain for synchronously treating municipal sewage and implementing grease accumulation |
| CN103571754A (en) * | 2013-05-09 | 2014-02-12 | 北京林业大学 | Chlorella Chlorella sp.HQ culture method and application in water purification and oil production |
| CN103952312A (en) * | 2013-12-31 | 2014-07-30 | 上海交通大学 | Freshwater Chlorella sorokiniana GS30 and application thereof |
| CN103952313A (en) * | 2013-12-31 | 2014-07-30 | 上海交通大学 | Imnetic algae strain Chlorella sorokiniana HN01 and application thereof |
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| CN104109634A (en) * | 2014-06-11 | 2014-10-22 | 武汉大学 | A construction method of a system for degrading azo dyes in water by algae microorganisms |
| CN105733951A (en) * | 2014-12-10 | 2016-07-06 | 中国农业大学 | A chlorella strain producing grease and applications thereof |
| CN105925484A (en) * | 2016-06-29 | 2016-09-07 | 广东海洋大学 | Normal temperature preservation method for oocystis |
| CN105985917A (en) * | 2015-02-06 | 2016-10-05 | 中国农业大学 | Method for improving biomass of chlorella in swine wastewater |
| CN107686815A (en) * | 2017-10-17 | 2018-02-13 | 中国水产科学研究院渔业机械仪器研究所 | A kind of method that chlorella is cultivated using food garbage |
| CN108359609A (en) * | 2018-03-15 | 2018-08-03 | 石家庄君乐宝乳业有限公司 | High survivability chlorella CV-6, its screening technique and its application |
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| CN110157622A (en) * | 2019-05-15 | 2019-08-23 | 中国辐射防护研究院 | A kind of screening technique of the chlorella algae strain for production of biodiesel |
| CN111500462A (en) * | 2020-04-13 | 2020-08-07 | 青岛旭能生物工程有限责任公司 | Method for industrial culture of nannochloropsis |
| CN113462577A (en) * | 2021-08-09 | 2021-10-01 | 中国科学院成都生物研究所 | Mutant chlorella and application thereof in sewage treatment |
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