CN104328053A - Scenedesmus capable of highly yielding oil as well as culture method and application thereof - Google Patents
Scenedesmus capable of highly yielding oil as well as culture method and application thereof Download PDFInfo
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Abstract
The invention relates to scenedesmus capable of highly yielding oil as well as a culture method and application thereof. An alga variety separated from a eutrophic water region is subjected to 18S rDNA sequence analysis and microstructure morphology identification so that the alga is determined as the member of Scenedesmus sp. and the sequence number is KF999043. The alga strain is named as Scenedesmus quadricauda SDEC-8 and is preserved in China Center for Type Culture Collection on October 8, 2014; and the preservation number is CCTCC NO: M2014448. An oil production culture method comprises the following steps: separating single alga cells of the Scenedesmus quadricauda SDEC-8 and carrying out aeration enlarged cultivation on obtained pure cells under laboratory conditions. The Scenedesmus quadricauda SDEC-8 contains rich high-quality oil; and the oil content can be up to 31.8% of the dry weight of the cells and the content of saturated fatty acids in fatty acids can be up to 80%, so that the Scenedesmus quadricauda SDEC-8 can be used as the raw material of biodiesel. Based on a high cetane number (61.38) and a low iodine number (29.38gI2/100g), the biodiesel prepared by taking the microalga as the raw material has good oxidization stability.
Description
Technical field
The invention belongs to technical field of microalga biology, particularly a kind of high yield oil fence algae and cultural method thereof and application.
Background technology
In recent years; because worldwide petrochemical rate of fuel consumption amount is increased sharply; the minimizing of petrochemical industry reserves; cause worldwide energy dilemma; add the environmental problem that fossil fuel burning and exhausting carbonic acid gas brings; therefore, the Renewable green energy finding a kind of cheapness has become Science & Society's problem of countries in the world government and common people's extensive concern.Research finds, it is high that some algae has oleaginousness, is easy to cultivate, yield per unit is large, do not strive with agricultural etc. advantage, be regarded as a new generation, or even uniquely can realize the biodiesel raw material of substitute fossil fuels.The lipid acid of its 16 carbon contained and 18 carbon is the most favourable for the production of biofuel.But production cost height is the principal element of restriction microalgae biodiesel scale production.And the algae kind that screening acquisition is easily cultivated, growth is fast, produce oil is high is the primary step overcoming micro-algae large-scale production bottleneck.As far back as the seventies in last century, the U.S. just starts the aquatic organism species project that cost 2,500 ten thousand dollars is operated by National Renewable Energy laboratory (NREL).NREL is separated to more than 3000 kind of algae from ocean and lake, therefrom filter out growth fast, comprise algae kind kinds more than 300 such as diatom, green alga and blue-green algae containing high, but unresolved Cost Problems at that time.The investigator of other various countries also utilizes various means to screen the excellent algae kind found and can adapt to commercially produce energetically in the world.The research that micro-algae prepares biofuel mainly concentrates on the following aspects: algae kind isolation and screening, algae culture, results and dehydration, grease extractive technique, algae bio converting fuel technology.Therefore, separation screening fast growth, high, the oily matter of fat content measured high-quality algae kind become the most important thing reducing biodiesel oil product cost from source.In recent years, domestic Duo Jia laboratory had started to carry out the work gathering and screen the strain of high produce oil algae on a large scale, and had made some progress.The micro-algae being selected from physical environment often has certain resistance in temperature, illumination and salinity adaptation, is suitable for mass propgation and produces.
Summary of the invention
Object of the present invention is just to overcome the problem for the production of the excellent algae strain deficiency of microalgae grease in prior art, a kind of high yield oil fence algae and cultural method thereof and application are provided, namely water sample is gathered at eutrophic water, a kind of oil-producing microalgae with resistance, easily open culture that separation and purification obtains, and set up cultural method.
For solving the problems of the technologies described above, the present invention is achieved by the following technical solutions:
The invention provides a kind of high yield oil fence algae, called after Scenedesmus quadricauda SDEC-8, be located away from the eutrophic lake of Quan Cheng park, Jinan City, be preserved in China typical culture collection center, its deposit number is: CCTCC NO:M 2014448.Preservation date: on October 8th, 2014.Address: Wuhan, China typical culture collection center (Wuhan University); Postcode: 430072; Deposit number: CCTCC NO:M 2014448.Scenedesmus quadricauda SDEC-8 is rich in high-quality grease, and fat content can reach 31.8% of dry cell weight, and in lipid acid, saturated fatty acid can reach 80%, can be used as biodiesel raw material.High hexadecane value (61.38) and low iodine number (29.38gI2/100g) determine with the micro-algae of this strain as biofuel prepared by raw material has good oxidation stability.
In an aspect, the invention provides a kind of Scenedesmus quadricauda SDEC-8 substratum, composition and consumption are NaNO
31500mgL
-1, K
2hPO
422.5 ~ 84.3mgL
-1, MgSO
47H
2o 75mgL
-1, CaCl
22H
2o 36mgL
-1, citric acid 6mgL
-1, ferric ammonium citrate 6mgL
-1, EDTANa
21mgL
-1, Na
2cO
320mgL
-1, H
3bO
32.86mgL
-1, MnCl
24H
2o 1.86mgL
-1, ZnSO
47H
2o 0.22mgL
-1, Na
2moO42H
2o 0.39mgL
-1, CuSO
45H
2o 0.08mgL
-1, Co (NO
3)
26H
2o 0.05mgL
-1, initial pH value is between 7 ~ 8, and postvaccinal algae cell density is 0.35 ~ 0.40 × 10
6cellsmL
-1.
Preferably, Media Components and consumption are: NaNO
31500mgL
-1, K
2hPO
440mgL
-1, MgSO
47H
2o75mgL
-1, CaCl
22H
2o 36mgL
-1, citric acid 6mgL
-1, ferric ammonium citrate 6mgL
-1, EDTANa
21mgL
-1, Na
2cO
320mgL
-1, H
3bO
32.86mgL
-1, MnCl
24H
2o 1.86mgL
-1, ZnSO
47H
2o 0.22mgL
-1, Na
2moO42H
2o 0.39mgL
-1, CuSO
45H
2o 0.08mgL
-1, Co (NO
3)
26H
2o 0.05mgL
-1, initial pH value=7.1, postvaccinal algae cell density is 0.37 × 10
6cellsmL
-1.
In one aspect of the method, the invention provides a kind of application of Scenedesmus quadricauda SDEC-8 substratum, for cultivating Scenedesmus quadricauda SDEC-8, obtaining Scenedesmus quadricauda SDEC-8 algae powder.
In an embodiment scheme, the fatty acid methyl ester 9.55mgg of described Scenedesmus quadricauda SDEC-8 algae powder
-1, saturated fatty acid content is 80%, and cetane value is 61.38, and iodine number is 29.38gI
2/ 100g.
Present invention also offers a kind of cultural method cultivating Scenedesmus quadricauda SDEC-8, comprise the steps:
A) be incubated at by Scenedesmus quadricauda SDEC-8 in the substratum adding BG11, culture temperature is 25 ± 1 DEG C, full exposure, intensity of illumination 2000-3000Lux, cultivates 7 days,
B) gained frustule is transferred in BG11 substratum carries out enlarged culturing, aeration vapour-liquid ratio is 0.1-0.2vvm, culture temperature is 25 ± 1 DEG C, full exposure, intensity of illumination 2000-3000Lux, centrifugal when Scenedesmus quadricauda SDEC-8 grows into two days stationary phases, be drying to obtain cultivate after Scenedesmus quadricauda SDEC-8 algae powder.The growth curve of Scenedesmus quadricauda SDEC-8 in BG11 substratum is shown in Fig. 3.The specific growth rate of Scenedesmus quadricauda SDEC-8 in standing BG11 nutrient solution and biomass concentration are respectively 0.17d
-1, 0.25gL
-1, and specific growth rate and biomass concentration are respectively 0.27d in aeration BG11 nutrient solution
-1, 1.29gL
-1.Therefore, in BG11 substratum, aeration greatly facilitates growth and the biomass accumulation of Scenedesmus quadricauda SDEC-8.
Preferably, step b) middle culture condition: culture temperature is 25 ± 1 DEG C, and illumination is the continuous illumination of 2500Lux, and aeration vapour-liquid ratio is 0.2vvm.Raising under the grease productivity ratio quiescent culture of Scenedesmus quadricauda SDEC-8 in the BG11 nutrient solution of aeration is more than 3 times, and fatty acid methyl ester productive rate improves nearly 9 times.Aeration greatly facilitates the oil and fat accumulation of grid algae SDEC-8.
We are bright provides the application of Scenedesmus quadricauda SDEC-8 in process eutrophic lake.Scenedesmus quadricauda SDEC-8 carries out the application of the purification of nitrogenous and/or phosphorus-containing wastewater.The production application of Scenedesmus quadricauda SDEC-8, for the production of the application of high-efficiency grease, lipid acid, protein, starch, pigment, polysaccharide and/or nucleic acid.Scenedesmus quadricauda SDEC-8 essentially comprising the lipid acid such as C14:0, C16:0, C18:0 and C18:1, and C16 and C18 system content is up to 96%, the wherein rich content of C16:0 and C18:1, analyzes from above lipid acid composition the requirement meeting preparation biofuel.
Beneficial effect of the present invention
The present invention sieves to obtain a strain advantage rich oil Scenedesmus quadricauda SDEC-8 (Scenedesmus quadricauda SDEC-8) in local polluted water body, the growth of this strain algae, without the need for the nutrition of machine matter, can obtain grease-contained biomass continuously by scale evaluation in the inorganic nutrients of cheapness.In its substratum, suitable aeration, can accelerate the mixing of algae liquid, improves the frustule efficiency of light energy utilization and growth velocity, Scenedesmus quadricauda SDEC-8 of the present invention under aeration clearly improve grease productive rate and biodiesel production rate, there is reasonable application prospect.Scenedesmus quadricauda SDEC-8 of the present invention (Scenedesmus quadricauda SDEC-8) is rich in grease, and has higher biofuel content than common algae, the biofuel character be more suitable for such as saturated fatty acid content up to 80%.The micro-algae of this strain is utilized to have good oxidation stability for biofuel prepared by raw material.
Accompanying drawing explanation
Fig. 1: Scenedesmus quadricauda SDEC-8 phylogenetic relationship tree in position.
The micromorphology photo of Fig. 2: Scenedesmus quadricauda SDEC-8
The pcr amplification product electrophoresis photographs of Fig. 3: 18S rRNA
Fig. 4: Scenedesmus quadricauda SDEC-8 in BG11 the growth curve of aeration and not aeration.
Wherein, 1 light micrograph, 6 μm, 2 scanning electron photomicrographs, scale, 3,4 transmission electron micrographs, scale 500nm.c, chloroplast(id); M, plastosome; N, nucleus; Py, pyrenoids; S, starch small grain; V, vacuole; A represents Scenedesmus quadricauda SDEC-8.
Embodiment
Mode by the following examples further illustrates the present invention, but does not therefore limit the present invention among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, conventionally selects with condition.
The isolation identification of embodiment 1 Scenedesmus quadricauda SDEC-8:
One, sample collecting
In July, 2012 contains the water sample of green alga from the eutrophic lake collection of Quan Cheng park, Jinan City, Shandong Province.
Two, the separation and purification of Scenedesmus quadricauda SDEC-8
Purification procedures: by blood cell plate counting process under the microscope to Botanical gardens water sample counting, liquid-tight for algae degree is diluted to about 3000/ml.On ultra-clean experiment table, BG11 solid culture liquid is down flat plate (placed and spent the night, without varied bacteria growing), algae liquid to be separated use laryngeal atomizer is sprayed on respectively on the flat board marked, then build, be placed in incubator and cultivate more than ten skies, the algal community of isolation mutually just can be there is on substratum, and by microscopy Chun Zao group, then be grafted directly to the transfering loop of sterilizing and nutrient solution be housed and in the 100ml Erlenmeyer flask of sterilizing, with gauze and sterile film sealing, cultivate.BG11; PH controls 7.1.Above-mentioned substratum as made semisolid medium, then adds agar 5 ~ 6 ‰.Examine under a microscope frustule whether to be purified, if having assorted algae by the further separation and purification of 96 orifice plate of the frustule improvement of enlarged culturing: shake well substratum, as frustule becomes polymer to make frustule colony break for unicellular as far as possible.Under the microscope with cell counting count board counting (again counting after need diluting when concentration is large), according to counted algae cell density, nutrient solution is diluted to every 50 μ l containing 0.9 frustule unit.After abundant mixing, draw 50 μ L nutrient solutions successively in 96 well culture plate holes with liquid-transfering gun, be placed with fresh BG11 nutrient solution (300-500 μ l) in each culture hole in advance, add a cover and with sealed membrane, culture plate is sealed, prevent from evaporating.Cultivation plate hole is cultivated in illumination box, after 6 ~ 12h, examines under inverted microscope and mark slender hilum.Slender hilum inner cell to be marked increase to more than 500 or there is color time, can switching enlarged culturing be carried out.Select the hole liquid-transfering gun of single cells grown to be transferred to by its culture in the culturing bottle of the fresh culture that 200mL is housed and carry out enlarged culturing.Obtain the pure algae strain of a strain, called after SDEC-8.
Three, the qualification of Scenedesmus quadricauda SDEC-8
1. the identification of morphology of Scenedesmus quadricauda SDEC-8
The algae of Scenedesmus quadricauda SDEC-8 falls morphological specificity: after cultivating on culture medium flat plate, can be observed green algae and fall, rounded, neat in edge, smooth, glossy.
The morphological features of Scenedesmus quadricauda SDEC-8: observe its form and meet Scenedesmus feature under microscope and scanning electron microscope, there is sheet chloroplast(id), frustule true property coenobium is flat, is made up of 2-4 cell, and colony's cell straight line arranged side by side forms a line; Cell Long Circle, avette, the upper and lower two ends of cell extensively circle, the straight or slightly bending thorn that outside colony, each tool in two ends up and down of cell is outwards oblique, cell walls is more level and smooth.Cell colony length and width depend on growth period, general long 9-26 μm, wide 8-14 μm.Transmission electron microscope observing result shows, and containing an independent chloroplast(id) in each frustule, and occupies half cell, comprises a pyrenoids clearly, by starch small grain bag quilt around pyrenoids in chloroplast(id).Vacuole not of uniform size can be found in cell, the irregular oily mater of Dark grey can be seen in some vacuoles.As Fig. 2;
2. the Molecular Identification of Scenedesmus quadricauda SDEC-8
(1) pcr amplification of 18S rRNA and order-checking:
Laboratory apparatus: compact centrifuge (Eppendorf, rotating speed >12000r/min); Electrophoresis apparatus (Liuyi Instruments Plant, Beijing); PCR thermal cycling amplification instrument (Eppendorf); Gel imaging instrument (Bio-Rad company of the U.S.).
Experimental technique: according to RNA isolation kit, centrifugal and extract the STb gene of algae kind from proper amount of fresh algae liquid, 18S rRNA amplification is carried out to DNA sample.Amplimer is as follows.
Forward primer is: 18 S-F (CCTGGTTGATCCTGCCAG);
Reverse primer is: 18 S-R (TTGATCCTTCTGCAGGTTCA).
PCR reaction is carried out in 50 μ L systems.Consisting of of reaction system: template DNA 1 μ L; 1.0U Taq PCR mixture; Forward primer 2 μ L; Reverse primer 0.5 μ L; Distilled water 20 μ L.
Pcr amplification condition: 94 DEG C of sex change 4min; 94 DEG C of 1min, 55 DEG C of 1min, 72 DEG C of 1.5min.Circulate 30 times; 72 DEG C extend 10min, 4 DEG C of placements.The 18S rRNA amplified production of sepharose with 1% to bacterial strain does electrophoresis detection, cuts adhesive tape after checking, reclaims test kit (Shanghai Sheng Gong biotechnology company limited) purified pcr product with DNA gel.Pcr amplification product after recovery entrusts Shanghai Sheng Gong biotechnology company limited to check order.
18S rRNA sequential analysis and Phylogenetic Analysis:
The pcr amplification product electrophoresis photographs of 18S rRNA as shown in Figure 3.By contrasting with both sides Marker, the fragment length of known target amplification product is about 1.8kb.
The 18S rRNA length obtaining Scenedesmus quadricauda SDEC-8 after order-checking is the sequence of 1054bp, be submitted to Genbank and (accession number is KF999643) is compared in other algae strains, find that the evolutionary distance of Scenedesmus quadricauda SDEC-8 and Scenedesmus sp. is the most close, determine that it belongs to Scenedesmus, identify in conjunction with microscopic morphology, preliminary identification its be Scenedesmus quadricauda, called after Scenedesmus quadricauda SDEC-8.
Based on the result of identification of morphology and Molecular Identification, SDEC-8 is accredited as Scenedesmus quadricauda (Scenedesmus quadricauda), be preserved in China typical culture collection center, its deposit number on October 8th, 2014: CCTCC NO:M 2014448.Address: Wuhan, China typical culture collection center (Wuhan University); Postcode: 430072.
The growth conditions optimization of embodiment 2 Scenedesmus quadricauda SDEC-8
In order to the growth making Scenedesmus quadricauda SDEC-8 faster and better, BG-11 medium base carries out training method, K
2hPO
4dosage (22.5 ~ 84.3mgL
-1), the Combinatorial Optimization of culture condition, whether aeration.
(1) substratum used is BG11
BG11 medium component is as shown in table 1, and before using, pH controls 7.1,120 DEG C, sterilizing 30 minutes.
(2) growing state of Scenedesmus quadricauda SDEC-8 in BG11
Scenedesmus quadricauda SDEC-8 cultivates activation after 7 days, prepare inoculation, and Scenedesmus quadricauda SDEC-8 algae liquid to be seeded needs first to use 15mgL through centrifugal (4000 revs/min × 10 minutes, at 4 DEG C)
-1naHCO
3solution rinses twice algae liquid.Then, algae kind will at the NaHCO of 1.0ml
3be inoculated in BG11 nutrient solution after resuspended in solution, move in 1L Erlenmeyer flask, sealing.Be placed in illumination box to cultivate, culture temperature is 25 ~ 30 DEG C, full exposure, intensity of illumination 2500Lux, leaves standstill and aerated culture.Every day carries out pH monitoring, and pH value fluctuates between 8-10.At Initial stage of culture, the pH of algae liquid comparatively initial value rises to some extent, and pH rises to about 10, higher at the 2 to 4 day, is up to 10.3, starts afterwards to reduce, and is in plateau gradually, about 9.
Result shows: the growth curve of Scenedesmus quadricauda SDEC-8 in BG11 substratum is shown in Fig. 4.The specific growth rate of Scenedesmus quadricauda SDEC-8 in standing BG11 nutrient solution and biomass concentration are respectively 0.17d
-1, 0.25gL
-1, and specific growth rate and biomass concentration are respectively 0.27d in aeration BG11 nutrient solution
-1, 1.29gL
-1.Therefore, in BG11 substratum, aeration greatly facilitates growth and the biomass accumulation of Scenedesmus quadricauda SDEC-8.
The produce oil characteristic of embodiment 3 Scenedesmus quadricauda SDEC-8
One, the cultivation of Scenedesmus quadricauda SDEC-8
According to condition and the culture medium culturing Scenedesmus quadricauda SDEC-8 of embodiment 2, be placed in illumination box and cultivate, culture temperature is 25 ~ 30 DEG C, full exposure, intensity of illumination 2500Lux, leaves standstill and aerated culture.The substratum used for BG11, BG11 medium component as shown in table 1, before using, pH controls 7.1,120 DEG C, sterilizing 30 minutes.
Two, the growth of Scenedesmus quadricauda SDEC-8 and grease yield are determined
1. Scenedesmus quadricauda SDEC-8 biomass estimation
By the observation to algae liquid cell density, after frustule growth phase enters stationary phase, algae liquid is gathered in the crops.Determine to gather in the crops volume, with 50ml centrifuge tube splendid attire algae liquid, put into large capacity refrigerated centrifuge and carry out centrifugal, remove supernatant liquor and retain bed mud Scenedesmus quadricauda, obtain algae powder at 60 DEG C of oven drying at low temperatures.Biomass calculates as equation (1):
In formula: m-algae dry weight, g;
V-algae liquid amasss, L.
2. the mensuration of Scenedesmus quadricauda SDEC-8 fat content
Take 0.1g dry algae powder in 50ml centrifuge tube, add 10ml chloroform/methanol (2:1) solution, with the broken 10min of Ultrasonic Cell Disruptor, then 4000r/min centrifugation 10min, retain organic phase, organic phase transferred in 60ml separating funnel, whole leaching process repeats twice.According to the organic phase volume be separated, add the sodium chloride solution (1:5) of 0.9%, fully shake up 1min, and stratification 15min, reclaim lower floor's grease extract solution and measure its volume, get 5ml and reclaim solution in 10ml glass test tube, dry up with nitrogen, Glass tubing is positioned in 80 DEG C of baking ovens and is dried to weight (about 30 minutes).Equation for Calculating fat content:
In formula: LW-based on the fat content under dry weight, g/g;
M
1-algae dried bean noodles weight, g;
M
0-10ml Glass tubing dry weight, g;
M
2-with the 10ml Glass tubing dry weight of grease, g;
The volume of V-low phase grease, ml.
Result is as shown in table 3, and acquired results of the present invention is for the raising under the grease productivity ratio quiescent culture of Scenedesmus quadricauda SDEC-8 in the BG11 nutrient solution of aeration is more than 3 times, and fatty acid methyl ester productive rate improves nearly 9 times.Aeration greatly facilitates the oil and fat accumulation of grid algae SDEC-8.
Table 3 Scenedesmus quadricauda SDEC-8 BG11 cultivate in grease productive rate and fatty acid methyl ester productive rate
3. Scenedesmus quadricauda SDEC-8 Fatty Acid Methyl Esters is analyzed
Take 0.1g algae powder (200mg) Duplicate Samples, put into centrifuge tube, add the solid acid of algae opaque amount 10%; Add methyl alcohol 2mL water-bath 40 minutes under 60 degree, then each sample adds the 4%KOH-methanol solution with methanol phase same volume 2ml, and water-bath 60 minutes; Mixed solution is poured into from centrifuge tube in the 10mL Glass tubing with a scale of having dried, add 1mL normal hexane, ultrasonication 10 minutes; After layering, get supernatant liquor 200 μ L in the tool plug glass tubule of having dried, add mark liquid (2mg/mL) in 25 μ L methyl margarates in each sample, sealing, get 1 μ L and carry out gas chromatography combined with mass spectrometry (GC-MS) mensuration; Remaining fatty acid methyl ester, shifts out with liquid-transfering gun, and metered volume.
Gas chromatographicanalyzer device: Trace GC-DSQII (thermoelectricity) gas chromatographicanalyzer; Chromatographic column: VF-23ms quartz capillary column, its specification is 30m × 0.25mm × 0.25 μm; Splitting ratio: 50:1; Injector temperature: 210 DEG C; Helium flow velocity: constant current, 1mL/min; Heating schedule:: 150 DEG C keep 1min, are raised to 165 DEG C with the speed of 1 DEG C of per minute; Ion source temperature: 250 DEG C; Sample size: l μ L; Data are by NIST mass spectrum database analysis.Adopt internal standard quanitation.
The fatty acid component of table 4 Scenedesmus quadricauda SDEC-8 in BG11
SFA=saturated fatty acid (14:0,16:0,18:0,24:0); MUFA=monounsaturated fatty acids (16:1,18:1); PUFA=polyunsaturated fatty acid (16:2,16:3,18:2,18:3,20:4,20:5,22:6).
Result shows that (as table 4) Scenedesmus quadricauda SDEC-8 essentially comprising the lipid acid such as C14:0, C16:0, C18:0 and C18:1, and C16 and C18 system content is up to 96%, the wherein rich content of C16:0 and C18:1, analyzes from above lipid acid composition the requirement meeting preparation biofuel.C16:0 aeration cultivate results Scenedesmus quadricauda SDEC-8 in content about 70%, be significantly higher than C16:0 content (Converti in the algae kind of other bibliographical informations, A., Casazza, A.A., Ortiz, E.Y., Perego, P., Del Borghi, M., 2009.Effect of temperature and nitrogen concentration on the growth and lipid content of Nannochloropsis oculata and Chlorella vulgaris for biodiesel production.Chem.Eng.Process.48,1146 – 1151).
4. Scenedesmus quadricauda SDEC-8 biofuel properties evaluations
Average degree of unsaturation according to following formulae discovery algae fatty acid methyl ester admixture:
ADU＝∑M╳Y
i
Wherein, ADU refers to the average degree of unsaturation of algae oil;
Y
irefer to the massfraction of each fatty acid component;
M is the number of carbon-carbon double bond in each fatty acid component.
And calculating biofuel character as viscosity, density, cloud point, cetane value, iodine number and calorific value by biofuel degree of unsaturation, their relation is respectively by following formulate.
y＝－0.6316x+5.2065
y＝0.0055x+0.8726
y＝－13.356x+19.994
y＝－6.6684x+62.876
y＝74.373x+12.71
y＝1.7601x+38.534
The biofuel character of table 5 Scenedesmus quadricauda SDEC-8 and the contrast of biofuel, ASTM biofuel and EN Biodiesel Standards
Result is as shown in table 5, by with U.S. ASTM D6751, the comparison of Europe EN 14214 biodiesel fuel quality standard and common biodiesel raw material oil mass range, draw under aeration and non-aeration two kinds of culture condition, the viscosity of Scenedesmus quadricauda SDEC-8 grease, density, cetane value and calorific value all meet these standards, and under two culture condition, biofuel nature difference is little.The high hexadecane value of this algae oil and low iodine number determine with the micro-algae of this strain as biofuel prepared by raw material has good oxidation stability.
By reference to the accompanying drawings the specific embodiment of the present invention is described although above-mentioned; but not limiting the scope of the invention; one of ordinary skill in the art should be understood that; on the basis of technical scheme of the present invention, those skilled in the art do not need to pay various amendment or distortion that creative work can make still within protection scope of the present invention.
Claims (10)
1. a high yield oil fence algae, it is characterized in that, be located away from eutrophic lake, called after Scenedesmus quadricauda SDEC-8, be preserved in China typical culture collection center on October 8th, 2014, its deposit number is: CCTCC NO:M 2014448.
2. a Scenedesmus quadricauda SDEC-8 substratum as claimed in claim 1, is characterized in that: composition and consumption are NaNO
31500mgL
-1, K
2hPO
422.5 ~ 84.3mgL
-1, MgSO
47H
2o 75mgL
-1, CaCl
22H
2o 36mgL
-1, citric acid 6mgL
-1, ferric ammonium citrate 6mgL
-1, EDTANa
21mgL
-1, Na
2cO
320mgL
-1, H
3bO
32.86mgL
-1, MnCl
24H
2o 1.86mgL
-1, ZnSO
47H
2o 0.22mgL
-1, Na
2moO42H
2o 0.39mgL
-1, CuSO
45H
2o 0.08mgL
-1, Co (NO
3)
26H
2o 0.05mgL
-1, initial pH value is between 7 ~ 8, and postvaccinal algae cell density is 0.35 ~ 0.40 × 10
6cellsmL
-1.
3. a Scenedesmus quadricauda SDEC-8 substratum as claimed in claim 2, is characterized in that: Media Components and consumption are NaNO
31500mgL
-1, K
2hPO
440mgL
-1, MgSO
47H
2o 75mgL
-1, CaCl
22H
2o 36mgL
-1, citric acid 6mgL
-1, ferric ammonium citrate 6mgL
-1, EDTANa
21mgL
-1, Na
2cO
320mgL
-1, H
3bO
32.86mgL
-1, MnCl
24H
2o 1.86mgL
-1, ZnSO
47H
2o 0.22mgL
-1, Na
2moO42H
2o 0.39mgL
-1, CuSO
45H
2o 0.08mgL
-1, Co (NO
3)
26H
2o 0.05mgL
-1, initial pH value=7.1, postvaccinal algae cell density is 0.37 × 10
6cellsmL
-1.
4. an application for Scenedesmus quadricauda SDEC-8 substratum as claimed in claim 3, is characterized in that, described culture medium culturing Scenedesmus quadricauda SDEC-8, obtains Scenedesmus quadricauda SDEC-8 algae powder.
5. apply as claimed in claim 4, it is characterized in that, the fatty acid methyl ester 9.55mgg of described Scenedesmus quadricauda SDEC-8 algae powder
-1, saturated fatty acid content is 80%, and cetane value is 61.38, and iodine number is 29.38gI
2/ 100g.
6. cultivate a cultural method of the Scenedesmus quadricauda SDEC-8 of claim 1, it is characterized in that, comprise the steps:
A) be incubated at by Scenedesmus quadricauda SDEC-8 in the substratum adding BG11, culture temperature is 25 ± 1 DEG C, full exposure, intensity of illumination 2000-3000Lux, cultivates 7 days;
B) gained frustule is transferred in BG11 substratum carries out enlarged culturing, aeration vapour-liquid ratio is 0.1-0.2vvm, culture temperature is 25 ± 1 DEG C, full exposure, intensity of illumination 2000-3000Lux, centrifugal when Scenedesmus quadricauda SDEC-8 grows into two days stationary phases, be drying to obtain cultivate after Scenedesmus quadricauda SDEC-8 algae powder.
7. method as claimed in claim 4, is characterized in that, step b) middle culture condition: culture temperature is 25 ± 1 DEG C, and illumination is the continuous illumination of 2500Lux, and aeration vapour-liquid ratio is 0.2vvm.
8. the application of Scenedesmus quadricauda SDEC-8 as claimed in claim 1 in process eutrophic lake.
9. Scenedesmus quadricauda SDEC-8 according to claim 1 carries out the application of the purification of nitrogenous and/or phosphorus-containing wastewater.
10. Scenedesmus quadricauda SDEC-8 according to claim 1 is for the production of the application of high-efficiency grease, lipid acid, protein, starch, pigment, polysaccharide and/or nucleic acid.
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| CN110093381A (en) * | 2019-04-23 | 2019-08-06 | 齐鲁工业大学 | A method of promote microalgae grease to accumulate using xylose |
| CN113237817A (en) * | 2021-05-07 | 2021-08-10 | 山东大学 | Depolymerization observation method and application of benthic cyanobacteria algae pad |
| CN114507602A (en) * | 2020-10-28 | 2022-05-17 | 中国石油化工股份有限公司 | Scenedesmus for producing oil and culture application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107177505A (en) * | 2016-03-11 | 2017-09-19 | 中国石油化工股份有限公司 | One plant of grid algae and its cultural method and application |
| CN107177505B (en) * | 2016-03-11 | 2020-11-10 | 中国石油化工股份有限公司 | Scenedesmus as well as culture method and application thereof |
| CN109136093A (en) * | 2018-08-27 | 2019-01-04 | 中国地质科学院岩溶地质研究所 | A kind of method of karst carbon remittance resource utilization |
| CN110093381A (en) * | 2019-04-23 | 2019-08-06 | 齐鲁工业大学 | A method of promote microalgae grease to accumulate using xylose |
| CN114507602A (en) * | 2020-10-28 | 2022-05-17 | 中国石油化工股份有限公司 | Scenedesmus for producing oil and culture application thereof |
| CN114507602B (en) * | 2020-10-28 | 2023-07-04 | 中国石油化工股份有限公司 | Scenedesmus oleander and culture application thereof |
| CN113237817A (en) * | 2021-05-07 | 2021-08-10 | 山东大学 | Depolymerization observation method and application of benthic cyanobacteria algae pad |
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