CN102453682B - Marine microalga and its culture method and application - Google Patents
Marine microalga and its culture method and application Download PDFInfo
- Publication number
- CN102453682B CN102453682B CN201010515598XA CN201010515598A CN102453682B CN 102453682 B CN102453682 B CN 102453682B CN 201010515598X A CN201010515598X A CN 201010515598XA CN 201010515598 A CN201010515598 A CN 201010515598A CN 102453682 B CN102453682 B CN 102453682B
- Authority
- CN
- China
- Prior art keywords
- isochrysis
- ccmm5001
- marine microalgae
- isochrysis galbana
- seawater
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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 relates to a marine microalga and its culture method and application. The marine microalga is isochrysis sp. CCMM5001, is preserved in the china center for type culture collection (CCTCC), and has a preservation number of CCTCC NO:M2010138. The culture method comprises separating a single algae cell of isochrysis sp. CCMM5001, carrying out enlargement culture of the single algae cell under laboratory conditions, carrying out centrifugation in a fourth day after an isochrysis sp. CCMM5001 growth platform stage, and then drying to obtain isochrysis sp. CCMM5001 powder. The isochrysissp. CCMM5001 powder can be utilized as a biodiesel raw material. The culture method of the marine microalga utilizes carbon dioxide in industrial exhaust gas thereby reducing greenhouse gas discharge, and realizes absorption of oxynitrides in industrial exhaust gas thereby reducing environmental pollution.
Description
Technical field
The present invention relates to marine microalgae, specifically a kind of marine microalgae and cultural method thereof and application.
Background technology
Marine microalgae is the main provider of organism and energy in the marine ecology system, compare with other raw material, have the photosynthetic efficiency height, adaptive capacity to environment is strong, growth cycle is short, not limited by weather, itself can biochemical Synthetic Oil, and fat content height, be easy to realize advantages such as scale operation, so marine microalgae shows wide application prospect as the new source for preparing biodiesel fuel.Exploitation marine microalgae resource is utilized seawater, non-farming soil and sun power, with simple mineral matter nutritional and CO
2Change into grease, extraordinary application development prospect is arranged, become one of science and technology competition hot issue in the world.This research starts from the forties in last century, and Germany cultivates little algae and therefrom seeks bio-oil materials as liquid fuel.Last century late nineteen seventies, USDOE has started one by National Renewable Energy Laboratory and has utilized the hydrobiont (little algae) of little algae production biofuel to plant plan, make great efforts collection through more than ten years and be separated to the little algae of 3000 strains, be stored in the University of Hawaii at present.The nineties, it is the project of earth research renewal technology plan that Japan internationality trade and the Department of Industry have subsidized a key name, filters out many strains and tolerates high CO
2Concentration, high temperature, fast growth, the algae kind of high-cell density be can form, the technology platform of photosynthetic organism reactor and the technical scheme of little algal biomass energy development set up.Since the nineties, Israel's sand desert survey studies show that all kinds of lipid acid of a kind of snowfield green alga accumulation can account for 50% of dry cell weight, wherein unsaturated fatty acids accounts for 80% of total fatty acids.U.S.'s the nineties has used modern biotechnology to develop the little ring of the little algae of transgenosis oceanographic engineering algae, lipid content surpasses 60% of dry weight under the laboratory condition, outdoor cultivation lipid output can reach more than 40% of dry weight, and theoretical calculate is produced 1～2.5 ton of diesel oil per year for every mu.2006, US Congress's Helios (TAIYANGSHEN) plan of 400,000,000 dollars of supports from sunlight to fuel of agreeing to furnish funds for, task was to realize the industrialization of algae produce oil by 2010, and the target of following production every day MMbbl bio-crude oil.
Domestic professors Wu Qingyu of Tsing-Hua University etc. serve as experiment algae kind with the green algate of fresh water chlorella, organic and the inorganic carbon of the application cell culture technique that takes the lead in (heterotrophic fermentation technology) control, the supply of nitrogen, acquisition chlorophyll disappears, the heterotrophism chlorella of cell variable color, compare with the chlorella of nourishing one's nature certainly of unconverted, the crude fat content of heterotrophism frustule has improved more than 4 times, but also exist because using the organic carbon source glucose amount to occur the problem of cost greatly, illustrate if improve the fat content of alga cells and just need research and propose the marine microalgae resource of cheap high oil content and more advanced technological method.Domestic biomass energy research contents mostly stresses the screening at biodiesel raw material, on the distribution of oil plant, selection, cultivation, genetic improvement and complete processing thereof and the equipment, does not relate to the marine microalgae biomass energy.
Summary of the invention
The object of the invention is to provide a kind of marine microalgae and cultural method and application.
For achieving the above object, the technical solution used in the present invention is:
A kind of marine microalgae: marine microalgae is Isochrysis galbana CCMM5001, and Isochrysis galbana CCMM5001 is preserved in Chinese typical culture collection center (CCTCC) on June 7th, 2010, and preserving number is CCTCC NO:M2010138.
With Isochrysis galbana (Isochrysis sp, CCMM5001) be incubated in the seawater that adds the L1 substratum, culture temperature is 20 ± 1 ℃, Light To Dark Ratio 12h: 12h, intensity of illumination 3000-4000Lux, cultivated 5-7 days, the gained frustule is transferred to carries out enlarged culturing in the L1 substratum, culture temperature is 20+1 ℃, Light To Dark Ratio 12h: 12h, intensity of illumination 3000-4000Lux, treat after the plateau of Isochrysis galbana growth the 4th day centrifugal, be drying to obtain and cultivate back Isochrysis galbana algae powder.
The seawater of described interpolation L1 substratum is with the equal high-temperature sterilization behind 0.45 μ m Fibrous membrane filtration of seawater, adds L1 nutritive salt, makes its salinity 30.
The application of marine microalgae: described Isochrysis galbana can be used as the raw material of biofuel.
The present invention has following advantage:
Marine microalgae provided by the present invention can be used as the raw material of production biofuel, the cultivation of marine microalgae of the present invention is the carbonic acid gas that utilizes in the industrial gaseous waste, alleviate the discharging of greenhouse gases, also can absorb the oxynitrides in the industrial gaseous waste, reduce environmental pollution; Its two, the productive rate of little algae unit surface exceeds tens of times of higher plants; Again, little algae does not have organ differentiation such as higher plant rhizome leaf, and under conditions such as nitrogen stress, some unicellular little algae can accumulate grease in a large number, and oleaginousness can be up to 30-70%; Its three, little algae can utilize beach, saltings, desert to carry out large scale culturing, can utilize seawater, salt buck and Desert Area underground water to cultivate, therefore little algae can not strive ground with farm crop, strives water.
Description of drawings
Fig. 1 drips distribution plan for CCMM5001 fat under the Electronic Speculum of the present invention.
Embodiment
Embodiment 1
Marine microalgae is Isochrysis galbana (Isochrysis sp.CCMM5001), is preserved in Chinese typical culture collection center (CCTCC) on June 7th, 2010, and preserving number is CCTCCNO:M2010138.
Isochrysis galbana (Isochrysis sp.CCMM5001), belong to Chrysophyta (chryspohyta) at little algae taxonomy, Chrysophyceae (chrysophyceae), Isochrysis galbana order (Iochysridales), Isochrysis galbana section (Isochysidacecae), Isochrysis galbana belongs to (Ischrysis Parke), Isochrysis galbana is exposed motor cell, and its cell shape is changeable, generally is oblong or ellipse, juvenile cell has a slightly flat dorsal ventral side, and Gu Cemianguan is oblong.The long 5-6 μ of general activity cell m, wide 2-4 μ m, thick 2.2-3 μ m, sporangiocyst diameter 5-6 μ m does not have cell walls, and two flagellums are isometric, and from the cell front end, its length is 1-2 times of cell.Usually cell movement is slow, approximately with the speed rotation of per second one rotation with circle round.When cell was static, flagellum was tetanic and fall down from health, but when motion, and flagellum is towards the place ahead of cell, and carried out the motion of fluctuation formula.
The cultural method of marine microalgae chrysophyceae:
At first, utilize kapillary choose from method (Chen Mingyao, biological feed cultivate, Beijing: Chinese agriculture press, 2001,71-73; G.M.Hallegraeff, D.M.Anderson, A.D.Cembella, H.O.Enevoldsen.Manual on Harmtul Marine Microalgae.Intergovernmental Oceanographic Commission.1995,45-50), isolate Isochrysis galbana (Isochrysis sp., single algae cell CCMM5001), cultivate in the seawater that adds the L1 substratum, culture dish is 96 orifice plates.The algal species cultivation temperature is 20 ± 1 ℃, Light To Dark Ratio 12h: 12h, intensity of illumination 3000-4000Lux.A clone clon that obtains is transferred in the larger container and cultivates after one week of cultivation, carries out enlarged culturing.
L1 substratum (Guillard and Hargraves, 1993.R.R.L.Guillard and P.E.Hargraves, Stichochrysis immobilis is a diatom, not a chrysophyte.Phycologia 32 (1993) pp.234-236.) comprises (1) nitrogen (NaNO
3, mother liquid concentration is 75g/L, being made into nutrient solution is 1ml NaNO
3Mother liquor: the 1L seawater of sterilizing); (2) phosphorus (NaH
2PO
4H
2The O mother liquid concentration is 5g/L, and being made into nutrient solution is 1ml NaH
2PO
4H
2O mother liquor: the 1L seawater of sterilizing); (3) micro-metals (L1 trace metal solution is made into nutrient solution and is 1ml L1 trace metal solution mother liquor: the 1L seawater of sterilizing); ((4)) VITAMIN (f/2 vitamin solution is made into nutrient solution and is 0.5ml f/2 vitamin solution mother liquor: the 1L seawater of sterilizing).
L1Medium
(Guillard?and?Hargraves?1993)
To?950mL?filtered?seawater?add：
L1Trace?Metal?Solution
(Guillard?and?Hargraves?1993)
To?950mL?dH
2O?add：
Make?final?volume?up?to?l?L?with?filtered?seawater.
f/2Vitamin?Solution
(Guillard&Ryther?1962，Guillard?1975)
To?950mL?dH
2O?add：
Secondly, carry out enlarged culturing.Be to cultivate above-mentioned purebred chrysophyceae 4000mL in the wide-necked bottle of 5000mL at volume, algae kind inoculum size is for adding 10% of L1 substratum seawater amount.Incubation time is one month.In the last week of cultivating, afterwards because the increasing of frustule quantity adopt with blood counting chamber counting, draw the growth curve of chrysophyceae with plant plankton tally counting every day.Known by the chrysophyceae growth curve of drawing, through 3 days lag phase, chrysophyceae grow into exponential phase of growth, reached plateau on the 24th day, and collected algae mud at the 4th day that is entering plateau: with nutrient solution with 3000rmin
-1, centrifugal 15min, remove supernatant liquor after, distilled water wash precipitation frond twice is again with 3000rmin
-1, centrifugal 15min removes supernatant liquor, collects algae mud, and vacuum lyophilization is standby in-20 ℃ of preservations afterwards.
Embodiment 2
The analysis of total fat percentage composition:
Get the cryodesiccated algae powder of above-mentioned gained Isochrysis galbana take by weighing certainweight (0.5～1g), add distilled water.Be placed on-20 ℃ down freezing, then in 40 ℃ of water-baths, dissolve 5min, freezing, dissolving is 2 times repeatedly, microscopically is observed broken situation (referring to Fig. 1).The dissolving back adds the 10ml volume ratio in algae liquid be 1: 2 anhydrous diethyl ether and the mixed solvent of sherwood oil, adds the KOH precipitation frustule of 5ml massfraction 10% behind the concussion mixing again, then at 4000rmin
-1Centrifugal 15min gets supernatant under the condition.60 ℃ of water-baths boil off unnecessary solvent rapidly to constant weight, and weighing gets thick fat content, calculate total fat percentage composition, (referring to table 1) with following formula.
Total fat percentage composition=(total fat/dried algae is heavy) * 100%
Table 1 chrysophyceae CCMM5001 total lipid content
Gained result of the present invention is that the thick fat content of Isochrysis galbana CCMM5001 is up to 45%, the highest with other class microalgae grease content of prior art, wherein at Sun Liqin (Sun Liqin, Yang Lintao, environmental factor is to the influence of Isochrysis galbana fatty acid content and composition, food research and development, 2006,27 (5): 11-13) within Bao Dao the 8.5%-46.3% scope, be higher than (Jiang Xiamin such as Jiang Xiamin, Zheng Yizhou, 14 kinds of little algae total lipid contents and lipid acid composition research, hydrobiont journal, 2003,27 (3): 243-247) survey (Wang Ming, Liu Ran, Xu Nins such as the thick fat content of 4 strain chrysophyceae 25%-33% and Wang Ming, Li Aifen, the fatty acid compositional analysis of the little algae of Duan Shunshan .13 kind. ecological science, 2006,25 (6): 542～544) the thick fat content of the Isochrysis galbana of surveying 19.3%.
Embodiment 3
The analysis of chrysophyceae fatty acid component
To add 5mL 0.4molL in the embodiment 2 dry above-mentioned thick fat samples
-1KOH-CH
3OH solution, 60 ℃ of heating in water bath saponification 2h are until the oil droplet completely dissolve.After the cooling, move in the centrifuge tube, (9: 1=commodity concentrated hydrochloric acid: distilled water) regulating pH to 1-2, add 1mL 5%NaCl solution again, then is 1: 4 CHCl with the 6ml volume ratio to add hydrochloric acid
3-C
6H
14Organic phase is got in the mixing solutions extraction.Adding 5mL massfraction is 14% BF in organic phase
3-CH
360 ℃ of esterification 1h of OH solution.Nitrogen dries up, and 2mL normal hexane constant volume is got 1mL solution in last sample bottle, carries out GC/MS and detects (referring to table 2).Laboratory apparatus is Agilent 7890N gas chromatograph-975C GC-MS (GC/MS).Chromatographic column is: HP-5MS, and (30m * 0.25mm 20 * 0.25um); Gas-chromatography operational condition: 250 ℃ of vaporizer temperature; 280 ℃ of transmission line temperature; The chromatographic column heating schedule: 150 ℃ of initial temperature rise to 280 ℃ with 10 ℃ of min-1, and keep 5min; Input mode: split stream sampling, splitting ratio are 100: 1; Sample size: 1 μ L.Retrieval NIST, the mass spectrum picture library is determined lipid acid kind in the sample, the method for definite employing area normalization of each lipid acid relative content is calculated.
Described KOH-CH
3OH is that KOH is dissolved in CH
3Among the OH, KOH is at CH
3Molar density among the OH is 0.4molL
-1Described BF
3-CH
3OH is BF
3Be dissolved in CH
3In the OH solution, its volume fraction is 14%.
The main lipid acid of table 2 CCMM5001 chrysophyceae algae strain is formed
Gained result of the present invention has comprised C basically for CCMM5001
14:0, C
16:0, C
18:1, C
18:4And C
22:5In lipid acid, and content is all higher, C
16And C
18Be content up to 85.27%, C wherein
16:0, C
16:1, C
18:1Content abundant, meet the requirement for preparing biofuel from molecular structure analysis.The grease composition of most of little algaes is mainly triglyceride level (〉=80%) and C
14-C
22Longer chain fatty acid (Metting F B, Biodiversity and application of microalgae, J Inorg Biochem, 1996,17 (5-6): 477-489), wherein lipid acid is with C
16With C
18Be that lipid acid is main (Guil-Guerrero etc., 2004), PUFAs is the main ingredient of unsaturated fatty acids, palmitinic acid is main ingredient (the Harwood J L of saturated fatty acid, Guschina I A, The versatility of algae and their lipid metabolism, Biochimie, 2009,91:679-684.).
Claims (4)
1. marine microalgae, it is characterized in that: marine microalgae is Isochrysis galbana Isochrysis sp.CCMM5001, is preserved in Chinese typical culture collection center (CCTCC) on June 7th, 2010, preserving number is CCTCC NO:M2010138.
2. cultural method by the described marine microalgae of claim 1, it is characterized in that: with Isochrysis galbana Isochrysis sp., CCMM5001 is incubated in the seawater that adds the L1 substratum, culture temperature is 20 ± 1 ℃, Light To Dark Ratio 12h: 12h, intensity of illumination 3000-4000Lux, cultivated 5-7 days, the gained frustule is transferred to carries out enlarged culturing in the L1 substratum, culture temperature is 20 ± 1 ℃, Light To Dark Ratio 12h: 12h, intensity of illumination 3000-4000Lux, treat after the plateau of Isochrysis galbana growth the 4th day centrifugal, be drying to obtain and cultivate back Isochrysis galbana algae powder.
3. by the cultural method of the described marine microalgae of claim 2, it is characterized in that: the seawater of described interpolation L1 substratum is with the equal high-temperature sterilization behind 0.45 μ m Fibrous membrane filtration of seawater, adds the L1 substratum, makes its salinity 30.
4. application by the described marine microalgae of claim 1, it is characterized in that: described Isochrysis galbana is as the raw material of biofuel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010515598XA CN102453682B (en) | 2010-10-15 | 2010-10-15 | Marine microalga and its culture method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010515598XA CN102453682B (en) | 2010-10-15 | 2010-10-15 | Marine microalga and its culture method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102453682A CN102453682A (en) | 2012-05-16 |
| CN102453682B true CN102453682B (en) | 2013-08-14 |
Family
ID=46037382
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010515598XA Active CN102453682B (en) | 2010-10-15 | 2010-10-15 | Marine microalga and its culture method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102453682B (en) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106164281A (en) * | 2013-08-23 | 2016-11-23 | 国立大学法人神户大学 | Produce method and the manufacture method of higher unsaturated fatty acid of lubricant component |
| CN104513800A (en) * | 2013-10-08 | 2015-04-15 | 镇江拜因诺生物科技有限公司 | Screening identification method for difficultly-cultured marine microorganisms |
| CN103966100B (en) * | 2014-05-26 | 2016-04-20 | 临沂大学 | The substratum of slaughterhouse's waste water cultivation Isochrysis galbana and different glue algae and cultural method |
| CN104560720B (en) * | 2014-12-12 | 2017-06-23 | 中国海洋大学 | Intend double spore algaes and its application and cultural method in one plant of ocean |
| CN105002094A (en) * | 2015-07-31 | 2015-10-28 | 临沂大学 | Method for semi-continuously culturing marine microalgae through solar greenhouse |
| CN106916748B (en) * | 2015-12-28 | 2020-06-16 | 国投生物科技投资有限公司 | Golden algae and its culture method |
| CN106190850B (en) * | 2016-07-08 | 2019-09-24 | 大连海洋大学 | The cultural method of high fat content microalgae body |
| CN106244489B (en) * | 2016-08-29 | 2020-02-04 | 佛山市艳晖生物科技有限公司 | Method for mixed fermentation of chrysophyceae and photosynthetic bacteria |
| CN107164271B (en) * | 2017-06-09 | 2020-01-14 | 中国科学院烟台海岸带研究所 | Marine microalgae and application thereof |
| CN107058114B (en) * | 2017-06-09 | 2020-01-10 | 中国科学院烟台海岸带研究所 | Marine electrogenesis microalgae and application thereof |
| CN107699538B (en) * | 2017-12-05 | 2020-11-03 | 杭州富阳优信科技有限公司 | Method for culturing Platymonas subcordiformis |
| CN110117543B (en) * | 2019-07-04 | 2021-04-30 | 杭州富阳优信科技有限公司 | Method for preparing algae powder by high-density culture of isochrysis galbana |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100473719C (en) * | 2006-01-27 | 2009-04-01 | 天津科技大学 | Outdoor large-area cultivating method for Isochrysis galbana H29 containing rich DHA |
-
2010
- 2010-10-15 CN CN201010515598XA patent/CN102453682B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100473719C (en) * | 2006-01-27 | 2009-04-01 | 天津科技大学 | Outdoor large-area cultivating method for Isochrysis galbana H29 containing rich DHA |
Non-Patent Citations (4)
| Title |
|---|
| 三种海洋微藻生长期脂肪酸组成研究;李文权等;《海洋环境科学》;20020531;第21卷(第2期);10-13 * |
| 李文权等.三种海洋微藻生长期脂肪酸组成研究.《海洋环境科学》.2002,第21卷(第2期),10-13. |
| 海洋微藻生产生物柴油的应用前景;韩笑天等;《海洋科学》;20081231;第32卷(第8期);76-81 * |
| 韩笑天等.海洋微藻生产生物柴油的应用前景.《海洋科学》.2008,第32卷(第8期),76-81. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102453682A (en) | 2012-05-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102453682B (en) | Marine microalga and its culture method and application | |
| Hossain et al. | Progress in physicochemical parameters of microalgae cultivation for biofuel production | |
| Sánchez et al. | Biomass resources | |
| Bharathiraja et al. | Aquatic biomass (algae) as a future feed stock for bio-refineries: A review on cultivation, processing and products | |
| John et al. | Micro and macroalgal biomass: a renewable source for bioethanol | |
| Sudhakar et al. | An overview of CO2 mitigation using algae cultivation technology | |
| CN102093955B (en) | Chlorella strain and application thereof | |
| Scarsella et al. | Study on the optimal growing conditions of Chlorella vulgaris in bubble column photobioreactors | |
| CN102036551A (en) | Algal culture production, harvesting, and processing | |
| CN104611228A (en) | Highly oil-containing monoraphidium and culture and application thereof | |
| CN103215190A (en) | Freshwater chlorella and application thereof in fixation of CO2 and production of microalgae oil | |
| CN101475823B (en) | Method for preparing biodiesel from sugarcane | |
| CN103266062B (en) | Fresh water Chlorella pyrenoidosa XJ01 strain and application thereof in fixing CO2 and producing microalga grease | |
| CN102492626A (en) | Nannochloropsis sp and application thereof | |
| Kings et al. | Growth studies on microalgae Euglena sanguinea in various natural eco-friendly composite media to optimize the lipid productivity | |
| Bhateria et al. | Algae as biofuel | |
| CN101386799B (en) | Production method of culture algae and refinery for coupling planting grass, power generation and heat supply | |
| Ievina et al. | Potential of Chlorella species as feedstock for bioenergy production: a review | |
| CN103842514B (en) | The fatty acid methyl ester (biodiesel) that can be used for electromotor of the marine microalgae cultivated in naturally occurring marine microalgae group and outdoor salt huge rock and the accretion of by-product | |
| CN103555584A (en) | Acquisition and application of grease-producing monoraphidium LB50 | |
| CN108456700A (en) | A method of improving grease in microalgae using give up mash and magnesium ion of molasses | |
| Cenciani et al. | Sustainable production of biodiesel by microalgae and its application in agriculture | |
| CN102911872B (en) | Scenedesmus sp. strain and application thereof | |
| CN102943044B (en) | Scenedesmus sp. and use thereof | |
| CN102433362A (en) | Method for producing biodiesel while treating biogas slurry by using microalgae |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |