CN109136093A - A kind of method of karst carbon remittance resource utilization - Google Patents
A kind of method of karst carbon remittance resource utilization Download PDFInfo
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- CN109136093A CN109136093A CN201810982312.5A CN201810982312A CN109136093A CN 109136093 A CN109136093 A CN 109136093A CN 201810982312 A CN201810982312 A CN 201810982312A CN 109136093 A CN109136093 A CN 109136093A
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 59
- 229910052799 carbon Inorganic materials 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 title claims abstract description 51
- 241000195493 Cryptophyta Species 0.000 claims abstract description 103
- 239000004519 grease Substances 0.000 claims abstract description 23
- 239000003225 biodiesel Substances 0.000 claims abstract description 16
- 238000012216 screening Methods 0.000 claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 6
- 238000011156 evaluation Methods 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 49
- 229910001868 water Inorganic materials 0.000 claims description 42
- 239000000243 solution Substances 0.000 claims description 36
- 238000005119 centrifugation Methods 0.000 claims description 30
- 239000000843 powder Substances 0.000 claims description 29
- 238000005286 illumination Methods 0.000 claims description 24
- 241000196324 Embryophyta Species 0.000 claims description 17
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 15
- OAYLNYINCPYISS-UHFFFAOYSA-N ethyl acetate;hexane Chemical compound CCCCCC.CCOC(C)=O OAYLNYINCPYISS-UHFFFAOYSA-N 0.000 claims description 15
- 239000006228 supernatant Substances 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 13
- 238000002360 preparation method Methods 0.000 claims description 12
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 12
- 238000000605 extraction Methods 0.000 claims description 11
- 239000013505 freshwater Substances 0.000 claims description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 10
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 8
- 239000000284 extract Substances 0.000 claims description 8
- 239000007836 KH2PO4 Substances 0.000 claims description 5
- 230000003698 anagen phase Effects 0.000 claims description 5
- 238000004364 calculation method Methods 0.000 claims description 5
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- 238000003113 dilution method Methods 0.000 claims description 5
- 229910052564 epsomite Inorganic materials 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- 230000000050 nutritive effect Effects 0.000 claims description 5
- 239000012074 organic phase Substances 0.000 claims description 5
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 5
- 150000003839 salts Chemical class 0.000 claims description 5
- 230000010355 oscillation Effects 0.000 claims description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 4
- 239000004575 stone Substances 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 238000013461 design Methods 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 claims 1
- 229910004619 Na2MoO4 Inorganic materials 0.000 claims 1
- 150000001336 alkenes Chemical class 0.000 claims 1
- 239000011684 sodium molybdate Substances 0.000 claims 1
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 9
- 230000008901 benefit Effects 0.000 abstract description 7
- 230000029553 photosynthesis Effects 0.000 abstract description 5
- 238000010672 photosynthesis Methods 0.000 abstract description 5
- 239000012620 biological material Substances 0.000 abstract description 4
- 238000005516 engineering process Methods 0.000 abstract description 2
- 241000195663 Scenedesmus Species 0.000 description 20
- 241000883966 Astrophytum capricorne Species 0.000 description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 13
- 239000002775 capsule Substances 0.000 description 9
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 8
- 241000192656 Nostoc Species 0.000 description 8
- 230000012010 growth Effects 0.000 description 7
- 239000011521 glass Substances 0.000 description 6
- 239000004698 Polyethylene Substances 0.000 description 5
- 230000009286 beneficial effect Effects 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- -1 polyethylene Polymers 0.000 description 5
- 229920000573 polyethylene Polymers 0.000 description 5
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 229960002413 ferric citrate Drugs 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- NPFOYSMITVOQOS-UHFFFAOYSA-K iron(III) citrate Chemical compound [Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NPFOYSMITVOQOS-UHFFFAOYSA-K 0.000 description 4
- 239000011435 rock Substances 0.000 description 4
- 239000000779 smoke Substances 0.000 description 4
- 238000005057 refrigeration Methods 0.000 description 3
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 2
- 241000192700 Cyanobacteria Species 0.000 description 2
- 240000002853 Nelumbo nucifera Species 0.000 description 2
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 2
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 2
- 240000001398 Typha domingensis Species 0.000 description 2
- 238000005260 corrosion Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000252229 Carassius auratus Species 0.000 description 1
- 229910015667 MoO4 Inorganic materials 0.000 description 1
- 240000008881 Oenanthe javanica Species 0.000 description 1
- 235000000365 Oenanthe javanica Nutrition 0.000 description 1
- 235000003283 Pachira macrocarpa Nutrition 0.000 description 1
- 235000014676 Phragmites communis Nutrition 0.000 description 1
- 235000013559 Schnittsellerie Nutrition 0.000 description 1
- 240000001085 Trapa natans Species 0.000 description 1
- 235000014364 Trapa natans Nutrition 0.000 description 1
- 241000746966 Zizania Species 0.000 description 1
- 235000002636 Zizania aquatica Nutrition 0.000 description 1
- 241001123263 Zostera Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 238000004177 carbon cycle Methods 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000002786 root growth Effects 0.000 description 1
- 235000009165 saligot Nutrition 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
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- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
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Abstract
The invention discloses a kind of methods of karst carbon remittance resource utilization, belong to karst carbon remittance application technology as the second resource field.The method of the karst carbon remittance resource utilization, includes the following steps: step 1: the screening of karst region microalgae；Step 2: microalgae oil-producing Performance Evaluation；Step 3: the conversion that karst carbon converges；Step 4: the Collection utilization of grease.The oil-producing microalgae that the present invention is screened using karst region is biomaterial, the karst Aquaponic microalgae rich in high concentration inorganic carbon generated using karstification, passes through the Atmospheric CO that algae photosynthesis is fixed by karstification2It is converted into the biodiesel with economic benefit, to realize remittance important breakthrough of the fundamental research to resource utilization of karst carbon.
Description
Technical field
The present invention relates to a kind of methods of karst carbon remittance resource utilization, belong to karst carbon remittance application technology as the second resource neck
Domain.
Background technique
Karstification is occurred in one of open system three-phase imbalance process.Using the whole world as scale, karstification
Be Global Carbon, water, calcium circulation a part, pass through lithosphere, hydrosphere, atmospheric thermodynamics couple three.When corrosion occurs for carbonate rock
When, karst shows as carbon remittance effect: Carbonate rock is maximum carbon storehouse on the earth,
The 99.55% of global total carbon is accounted for, is the 1694 and 1.1 × 10 of ocean and land vehicles total carbon respectively6Times.Modern Karstology
Studies have shown that karstification plays an active part in global carbon.Carbonate rock area is 2.2 × 10 in the world7km2, account for about
The 15% of land area.China is karst big country, area about 3.44 × 106km2, account for about the 1/3 of national territorial area.Pass through the world
Geology comparison plan (International Geological Correlation Programmes, IGCP) is studies have shown that complete
Ball and China are every year because Atmospheric CO is recycled in carbonate rock corrosion2Amount be respectively 2.2 × 108-6.08×108Ton and 1.77 × 107
Ton.The karst water for carrying a large amount of inorganic carbon has significant facilitation to the growth of aquatic microalgae.It is single under karst water environment
Raw egg capsule algae utilizes HCO3 -Ability it is 4.78 times higher than non-karst water environment.Therefore, karst water is that the growth of microalgae is created well
Environment.However for many years in terms of the research of karst carbon cycle is only limitted to basic theory, in karst carbon remittance resource utilization
Aspect lacks breakthrough.
The research that overseas utilization microalgae prepares biodiesel is more early, wherein larger research be 1978-1996 by
" Aquatic Species Program (ASP) " project that U.S. Department of Energy is subsidized, nearest U.S. Department of Energy have subsidized one again
The fund of the project of entitled " Biomass Program ", the project 19% is developed for algae bio diesel oil.Japanese government also provides
Having helped a key name is the project of " earth study update method plan ", which invests about 2,500,000,000 dollars altogether, filter out more plants it is resistance to
By high CO2The algae that concentration, the speed of growth are fast, biomass is high.Nearly 2 years many production of energy giant company such as shell in the world
Petroleum, British Petroleum Company, Exxon Mobil etc. are all added in the research and development ranks of the microalgae energy and have put into a large amount of moneys
Gold.Microalgae has many advantages, such as that short growth cycle, easy industrialization culture and lipid material content are high, is prepare biodiesel good
Good material has become current research hotspot with microalgae preparation biodiesel.But currently, there is not yet utilizing karst in global range
Carbon banknotes-system for biodiesel relevant report.
Summary of the invention
Present invention aims to solve the deficiencies of the prior art, and provides a kind of a kind of methods of karst carbon remittance resource utilization.This
The oil-producing microalgae that invention is screened using karst region utilizes the karst rich in high concentration inorganic carbon of karstification generation as biomaterial
Aquaponic microalgae passes through the Atmospheric CO that algae photosynthesis is fixed by karstification2It is converted into the biological bavin with economic benefit
Oil, to realize remittance important breakthrough of the fundamental research to resource utilization of karst carbon.
The technical scheme to solve the above technical problems is that a kind of method of karst carbon remittance resource utilization, packet
Include following steps:
Step 1: the screening of karst region microalgae
The microalgae in karst aquifer is collected, then takes former karst water locating for the microalgae, for impregnating the microalgae, is protected in 4 DEG C
It deposits, obtains algae stoste；
Using concentration gradient dilution method, algae stoste is isolated and purified, obtains multiple pure strains, and determine its kind respectively
Belong to；
Step 2: microalgae oil-producing Performance Evaluation
Multiple pure strains that step 1 is obtained are respectively placed in its corresponding culture solution, are carried out illumination constant temperature and are expanded training
It supports, frustule, then vacuum freeze drying is collected by centrifugation, obtains the freeze-dried algae powder of multiple pure strains respectively；
Ethyl acetate-hexane method is assisted using ultrasonic wave, extracts the grease in above-mentioned multiple pure strain freeze-dried algae powders, and
The oil productivity for calculating separately above-mentioned multiple pure strains filters out the highest algae of oil productivity, is named as algae K；
Step 3: the conversion that karst carbon converges
Karst water is taken, is filtered, nutritive salt is added in smoke filtrate, preparation obtains the culture solution of microalgae kind K；
The algae K for taking the step 2 in logarithmic growth phase to obtain, 10%-15% is inoculated in the training of microalgae kind K by volume
In nutrient solution, carries out illumination constant temperature and expand culture, frustule, then vacuum freeze drying is collected by centrifugation, obtains the freeze-drying algae of algae K
Powder；
Step 4: the Collection utilization of grease
Ethyl acetate-hexane method is assisted using ultrasonic wave, the oil in the freeze-dried algae powder for the algae K that extraction step 3 obtains
Rouge, and it is used to prepare biodiesel.
The principle of the present invention:
Water is the important intermediate medium of KARST ECOSYSTEM, and karstification is by atmosphere/soil CO of absorption2With HCO3 -'s
Form brings water body into improve the inorganic carbon content of karst aquifer, and the karst water energy rich in high concentration inorganic carbon greatly facilitates
The growth and breeding of microalgae cell.Therefore, atmosphere/soil CO using the photosynthesis of oil-producing microalgae that karstification is fixed2
The organic matter of itself is converted into fixed carbon, so that carbonizable substance is converted into grease by cell cometabolism process, finally
Grease is collected using the method for physics, chemistry, for large scale preparation biodiesel.
In step 1 of the invention, the identification of the kind of pure strain, referring to " CHINESE FRESHWATER algae --- genealogical classification and life
State " (Hu Hongjun, Wei Yin heart CHINESE FRESHWATER algae --- genealogical classification and the Beijing ecology [M]: Science Press, 2006).
In step 2 of the invention, the corresponding culture solution of pure strain, referring to " Chinese Academy of Sciences's fresh water algae library "
(http://algae.ihb.ac.cn/Default.aspx)。
Based on the above technical solution, the present invention can also be improved as follows.
Further, described to collect the specific method of microalgae in karst aquifer and be in step 1: in karst aquifer, with swimming
Biological net draws " ∞ " word, either scrapes microalgae from submerged plant blade face, emergent aquactic plant stem table or stone with banister brush, then use
After planktonic organism net filtration, it is transferred in polyethylene bottle.
It is using above-mentioned further beneficial effect: may be implemented three-dimensional, complete from the upper, middle and lower-ranking of karst aquifer
The aquatic microalgae of the collection in face.
Above-mentioned submerged plant refers to that plant is entirely located in the big shape water plant that fixed life is sought below water layer.They
Root it is sometimes undeveloped or degenerate, all absorbable moisture of each section of plant and nutriment, aerating tissue is especially flourishing, is conducive to
Gas exchanges are carried out in the case where lacking air in water.The leaf of this kind of plant be mostly it is band-like or Filamentous, such as eel grass, goldfish
Algae, watermifoil, black algae etc..
Among the bed mud of water, stem, leaf very go out the water surface for emergent aquactic plant, the i.e. root, root growth of plant；It is often distributed in 0-
The shoal of 1.5m, wherein some types are grown on moist bank.This kind of aerial part of plant has terrestrial plant
Feature；The part (root or subterranean stem) of growth in water, the feature with water plant.It is common have reed, the stem or leaf of cattail, water chestnut, lotus,
Chinese celery, wild rice stem Xun, lotus, cattail etc..
In general, being saved prior to 4 DEG C after polyethylene bottle is collected into microalgae, then transporting laboratory back rapidly, carried out subsequent
Operation.
Further, the aperture of the plankton net is No. 25.Aperture is No. 25, i.e., aperture is 64 μm.
Further, in step 2, the corresponding culture solution, according to " Chinese Academy of Sciences's fresh water algae library ", network address is
Http:// algae.ihb.ac.cn/Default.aspx is carried out.
Further, in step 2, the calculation method of the oil productivity are as follows: oil productivity=M/N × 100%, in formula, N is freeze-drying
The quality (g) of algae powder；M is the quality (g) of grease.
Further, in step 2 and step 3, the speed of the centrifugation is 10000-12000r/min, and the time of centrifugation is
10min；The temperature of the vacuum freeze drying is -55 DEG C, vacuum degree 5Pa, time 12h.
It is using above-mentioned further beneficial effect: using above-mentioned parameter, can sufficiently collects frustule, and keeps it de-
Water is complete, convenient for the extraction of subsequent grease.
Further, in step 3, the content of inorganic carbon is 3-5mmol/L in the karst water；The design parameter of the suction filtration
Are as follows: the aperture of filter membrane is 0.22 μm, pressure 0.098MPa, sucking rate 10L/min；The culture solution of the microalgae kind K is 1L body
Contain the NaNO of 1.5g in product3, 0.04g KH2PO4, 0.075g MgSO4·7H2O, the CaCl of 0.036g2·2H2O、
The citric acid of 0.006g, the ferric citrate of 0.006g, 0.001g EDTANa2, 0.02g Na2CO3, 1mL the micro member of A5
Plain liquid, surplus are karst water.
It is using above-mentioned further beneficial effect: the culture solution of microalgae kind K, with the prior art the difference is that adopting
Distilled water is replaced with karst water, obtained culture solution has the growth of microalgae kind K due to being rich in high concentration inorganic carbon great
Facilitation.
Further, the group of the A5 liquid microelement becomes the H of 2.86g/L3BO3, the MnCl of 1.86g/L2·4H2O,
The ZnSO of 0.22g/L4·7H2The Na of O, 0.39g/L2MoO4·2H2The CuSO of O, 0.08g/L4·5H2The Co of O and 0.05g/L
(NO3)2·6H2O。
Be using above-mentioned further beneficial effect: A5 trace element solution is the weight of BG-11 culture medium microelement
Want component part.BG-11 is to cultivate the most popular culture medium of cyanobacteria can largely use BG- in addition to only a few cyanobacteria
11 culture mediums.
Further, in step 2 and step 4, the specific method of the ultrasonic wave auxiliary ethyl acetate-hexane method is: point
50mg ± 2mg freeze-dried algae powder is also known as taken, broken wall is ground, be transferred to centrifuge tube with cover and the n-hexane that 5mL volume ratio is 1:1 is added
With the mixed liquor of ethyl acetate, after sonic oscillation 15min, 50 DEG C of water-bath 45min vibrate 3 times therebetween, 10000r/min centrifugation,
Supernatant is collected, primary extract is completed；Liquid Residue is repeated into extraction 2 times, merges supernatant；The bodies such as combined supernatant are added
Long-pending deionized water, and be uniformly mixed, 2000r/min centrifugation takes upper organic phase, is dried with nitrogen to constant weight.
Further, in step 2 and step 4, the temperature that the illumination constant temperature expands culture is 25 DEG C, and the time is 7 days.
Beneficial effects of the present invention:
1, the present invention is rich in high concentration using what karstification generated using the oil-producing microalgae of karst region screening as biomaterial
The karst Aquaponic microalgae of inorganic carbon passes through the Atmospheric CO that algae photosynthesis is fixed by karstification2Being converted into has economy
The biodiesel of benefit, to realize remittance important breakthrough of the fundamental research to resource utilization of karst carbon.
2, the carbon Correspondent that karstification generates is crossed the photosynthetic of algae by the method for karst carbon of the invention remittance resource utilization
Effect is largely converted into the raw material of preparation biodiesel.And biodiesel belongs to environmentally friendly green fuel, and therefore, this hair
Bright method has far-reaching economic benefit and social benefit.
3, karst carbon of the invention converges the method for resource utilization, is at home and abroad put forward for the first time, realize karst carbon converge from
Important breakthrough of the theoretical research to resource utilization.
4, the method for karst carbon of the invention remittance resource utilization, easy to operate, wide market, suitable scale push away
Wide application.
Specific embodiment
Principles and features of the present invention are described below in conjunction with specific embodiment, example is served only for explaining this hair
It is bright, it is not intended to limit the scope of the present invention.
Embodiment 1
The method of the karst carbon remittance resource utilization of the present embodiment, includes the following steps:
Step 1: the screening of karst region microalgae
In typical ocean-stockaded village bottom Underground river system supply Karst geomorphy korneforos, " ∞ " word is drawn with plankton net, is scraped
Microalgae is taken, then after the planktonic organism net filtration for being No. 25 with aperture, is transferred in polyethylene bottle, then take protolith locating for the microalgae
Molten water saves in 4 DEG C for impregnating the microalgae, obtains algae stoste.
Using concentration gradient dilution method, algae stoste is isolated and purified, referring to " CHINESE FRESHWATER algae --- genealogical classification
And ecology " algae of screening is identified, it is determined as chlorella and scenedesmus.
Step 2: microalgae oil-producing Performance Evaluation
According to " Chinese Academy of Sciences's fresh water algae library ", network address is http://algae.ihb.ac.cn/Default.aspx,
The corresponding culture solution BG-11 of chlorella is prepared, chlorella is placed in culture solution BG-11, illumination constant temperature is carried out and expands culture,
The intensity of illumination is 3000lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Preparing the corresponding culture solution of scenedesmus is BG-11, and scenedesmus is placed in culture solution BG-11, carries out the expansion of illumination constant temperature
Big culture, the intensity of illumination are 2500lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Frustule is collected by centrifugation, the speed of centrifugation is 10000r/min, and the time of centrifugation is 10min, then vacuum refrigeration is done
Dry, the temperature of vacuum freeze drying is -55 DEG C, vacuum degree 5Pa, time 12h, obtains the freeze-drying of chlorella and scenedesmus respectively
Algae powder.
Ethyl acetate-hexane method is assisted using ultrasonic wave, extracts the grease in the freeze-dried algae powder of chlorella and scenedesmus.Institute
The specific method for stating ultrasonic wave auxiliary ethyl acetate-hexane method is: 500mg ± 2mg (being calculated as M) freeze-dried algae powder is weighed respectively,
Broken wall is ground, centrifuge tube with cover is transferred to and the mixed liquor of n-hexane and ethyl acetate that 5mL volume ratio is 1:1, ultrasound is added
After vibrating 15min, 50 DEG C of water-bath 45min vibrate 3 times therebetween, and supernatant is collected in 10000r/min centrifugation, complete primary extract；
Liquid Residue is repeated into extraction 2 times, merges supernatant；The isometric deionized water of combined supernatant is added, and is uniformly mixed,
2000r/min centrifugation, takes upper organic phase, is transferred in the glass tube weighed and (is calculated as N1), it is dried with nitrogen to constant weight, again
It weighs glass tube weight and (is calculated as N2)。
Calculate separately the oil productivity of above-mentioned chlorella and scenedesmus, the calculation method of oil productivity are as follows: oil productivity=M/N ×
100%, in formula, N is the quality (g) of freeze-dried algae powder, N=N2-N1；M is the quality (g) of grease.It is calculated:
The oil productivity of chlorella=(20.2421-20.0907)/0.5 × 100%=30.28%；
The oil productivity of scenedesmus=(20.2512-20.0908)/0.5 × 100%=32.08%.
In chlorella and scenedesmus, the oil productivity of scenedesmus is relatively higher.
Step 3: the conversion that karst carbon converges
Karst water is taken, the content of inorganic carbon is 3mmol/L in karst water；It filters, the aperture of filter membrane is 0.22 μm, and pressure is
0.098MPa, sucking rate 10L/min, are added nutritive salt in smoke filtrate, and preparation obtains the culture solution of scenedesmus；The culture solution of scenedesmus
For the NaNO containing 1.5g in 1L volume3, 0.04g KH2PO4, 0.075g MgSO4·7H2O, the CaCl of 0.036g2·
2H2O, the EDTANa of the citric acid of 0.006g, the ferric citrate of 0.006g, 0.001g2, 0.02g Na2CO3, the A5 of 1mL it is micro-
Secondary element liquid, remaining is karst water.
The scenedesmus in logarithmic growth phase is taken, 10% is inoculated in the BG-11 culture solution of karst water preparation by volume, into
Row illumination constant temperature expands culture, and the intensity of illumination is 2500lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Frustule, then vacuum freeze drying is collected by centrifugation, obtains the scenedesmus freeze-dried algae powder of karst Aquaponic；
Step 4: the Collection utilization of grease
Ethyl acetate-hexane method is assisted using ultrasonic wave, the scenedesmus that concrete operations are obtained with step 2, extraction step 3
Grease in freeze-dried algae powder, and it is used to prepare biodiesel.
Embodiment 2
The method of the karst carbon remittance resource utilization of the present embodiment, includes the following steps:
Step 1: the screening of karst region microalgae
Typical hair village Karst subterranean river systems system supply Karst geomorphy korneforos, with banister brush from submerged plant blade face,
Microalgae is scraped on emergent aquactic plant stem table or stone, then after the planktonic organism net filtration for being No. 25 with aperture, is transferred to polyethylene
In bottle, then former karst water locating for the microalgae is taken, for impregnating the microalgae, is saved in 4 DEG C, obtain algae stoste.
Using concentration gradient dilution method, algae stoste is isolated and purified, referring to " CHINESE FRESHWATER algae --- genealogical classification
And ecology " algae of screening is identified, it is determined as chroococcoid and nostoc.
Step 2: microalgae oil-producing Performance Evaluation
According to " Chinese Academy of Sciences's fresh water algae library ", network address is http://algae.ihb.ac.cn/Default.aspx,
The corresponding culture solution BG-11 of chroococcoid is prepared, chroococcoid is placed in culture solution BG-11, illumination constant temperature is carried out and expands culture,
The intensity of illumination is 2000lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Preparing the corresponding culture solution of nostoc is BG-11, and nostoc is placed in culture solution BG-11, and it is permanent to carry out illumination
Temperature expands culture, and the intensity of illumination is 2000lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Frustule is collected by centrifugation, the speed of centrifugation is 10000r/min, and the time of centrifugation is 10min, then vacuum refrigeration is done
Dry, the temperature of vacuum freeze drying is -55 DEG C, vacuum degree 5Pa, time 12h, obtains the jelly of chroococcoid and nostoc respectively
Dry algae powder.
Ethyl acetate-hexane method is assisted using ultrasonic wave, extracts the grease in the freeze-dried algae powder of chroococcoid and nostoc.
The specific method of the ultrasonic wave auxiliary ethyl acetate-hexane method is: weighing 500mg ± 2mg (being calculated as M) freeze-drying algae respectively
Powder grinds broken wall, is transferred to centrifuge tube with cover and the mixed liquor of n-hexane and ethyl acetate that 5mL volume ratio is 1:1 is added, surpass
After sound oscillation 15min, 50 DEG C of water-bath 45min vibrate 3 times therebetween, and supernatant is collected in 10000r/min centrifugation, and completion once mentions
It takes；Liquid Residue is repeated into extraction 2 times, merges supernatant；The isometric deionized water of combined supernatant is added, and mixes equal
Even, 2000r/min centrifugation takes upper organic phase, is transferred in the glass tube weighed and (is calculated as N1), it is dried with nitrogen to constant weight,
Glass tube weight is weighed again (is calculated as N2)。
Calculate separately the oil productivity of above-mentioned chroococcoid and nostoc, the calculation method of oil productivity are as follows: oil productivity=M/N ×
100%, in formula, N is the quality (g) of freeze-dried algae powder, N=N2-N1；M is the quality (g) of grease.It is calculated:
The oil productivity of chroococcoid=(20.1976-20.0910)/0.5 × 100%=21.32%；
The oil productivity of nostoc=(20.1856-20.0907)/0.5 × 100%=18.98%.
In chroococcoid and nostoc, the oil productivity of chroococcoid is relatively higher.
Step 3: the conversion that karst carbon converges
Karst water is taken, the content of inorganic carbon is 4mmol/L in karst water；It filters, the aperture of filter membrane is 0.22 μm, and pressure is
0.098MPa, sucking rate 10L/min, are added nutritive salt in smoke filtrate, and preparation obtains the culture solution of scenedesmus；The culture solution of scenedesmus
For the NaNO containing 1.5g in 1L volume3, 0.04g KH2PO4, 0.075g MgSO4·7H2O, the CaCl of 0.036g2·
2H2O, the EDTANa of the citric acid of 0.006g, the ferric citrate of 0.006g, 0.001g2, 0.02g Na2CO3, the A5 of 1mL it is micro-
Secondary element liquid, remaining is karst water.
The chroococcoid in logarithmic growth phase is taken, 10% is inoculated in the BG-11 culture solution of karst water preparation by volume,
It carries out illumination constant temperature and expands culture, the intensity of illumination is 2500lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Frustule, then vacuum freeze drying is collected by centrifugation, obtains the chroococcoid freeze-dried algae powder of karst Aquaponic；
Step 4: the Collection utilization of grease
Ethyl acetate-hexane method, the chroococcoid that concrete operations are obtained with step 2, extraction step 3 are assisted using ultrasonic wave
Freeze-dried algae powder in grease, and be used to prepare biodiesel.
Embodiment 3
The method of the karst carbon remittance resource utilization of the present embodiment, includes the following steps:
Step 1: the screening of karst region microalgae
In the Guilin City test site Ya Ji karst subterranean stream exit, " ∞ " word is drawn with plankton net, scrapes microalgae, then use
After the planktonic organism net filtration that aperture is No. 25, it is transferred in polyethylene bottle, then take former karst water locating for the microalgae, for soaking
The microalgae is steeped, is saved in 4 DEG C, obtains algae stoste.
Using concentration gradient dilution method, algae stoste is isolated and purified, referring to " CHINESE FRESHWATER algae --- genealogical classification
And ecology " algae of screening is identified, it is determined as single raw egg capsule algae and goat's horn crescent moon algae.
Step 2: microalgae oil-producing Performance Evaluation
According to " Chinese Academy of Sciences's fresh water algae library ", network address is http://algae.ihb.ac.cn/Default.aspx,
The corresponding culture solution BG-11 of single raw egg capsule algae is prepared, single raw egg capsule algae is placed in culture solution BG-11, illumination constant temperature is carried out
Expand culture, the intensity of illumination is 5000lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Preparing the corresponding culture solution of goat's horn crescent moon algae is BG-11, and goat's horn crescent moon algae is placed in culture solution BG-11, into
Row illumination constant temperature expands culture, and the intensity of illumination is 4000lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Frustule is collected by centrifugation, the speed of centrifugation is 10000r/min, and the time of centrifugation is 10min, then vacuum refrigeration is done
Dry, the temperature of vacuum freeze drying is -55 DEG C, vacuum degree 5Pa, time 12h, obtains single raw egg capsule algae and the goat's horn moon respectively
The freeze-dried algae powder of tooth algae.
Ethyl acetate-hexane method is assisted using ultrasonic wave, extracts the freeze-dried algae powder of single raw egg capsule algae and goat's horn crescent moon algae
In grease.The specific method of the ultrasonic wave auxiliary ethyl acetate-hexane method is: weighing 500mg ± 2mg respectively and (is calculated as
M) freeze-dried algae powder grinds broken wall, is transferred to centrifuge tube with cover and n-hexane and ethyl acetate that 5mL volume ratio is 1:1 is added
Mixed liquor, after sonic oscillation 15min, 50 DEG C of water-bath 45min vibrate 3 times therebetween, and supernatant is collected in 10000r/min centrifugation, complete
It is extracted at primary；Liquid Residue is repeated into extraction 2 times, merges supernatant；The isometric deionized water of combined supernatant is added,
And be uniformly mixed, 2000r/min centrifugation takes upper organic phase, is transferred in the glass tube weighed and (is calculated as N1), it is dried with nitrogen
To constant weight, glass tube weight is weighed again and (is calculated as N2)。
Calculate separately the oil productivity of above-mentioned single raw egg capsule algae and goat's horn crescent moon algae, the calculation method of oil productivity are as follows: oil productivity
=M/N × 100%, in formula, N is the quality (g) of freeze-dried algae powder, N=N2-N1；M is the quality (g) of grease.It is calculated:
Oil productivity=(20.1082-20.0906)/0.5 × 100%=3.52% of single raw egg capsule algae；
The oil productivity of goat's horn crescent moon algae=(20.1645-20.0907)/0.5 × 100%=14.76%.
Single raw egg capsule algae and goat's horn crescent moon algae, the oil productivity of goat's horn crescent moon algae are relatively higher.
Step 3: the conversion that karst carbon converges
Karst water is taken, the content of inorganic carbon is 5mmol/L in karst water；It filters, the aperture of filter membrane is 0.22 μm, and pressure is
0.098MPa, sucking rate 10L/min, are added nutritive salt in smoke filtrate, and preparation obtains the culture solution of BG-11；Goat's horn crescent moon algae
Culture solution be 1L volume in the NaNO containing 1.5g3, 0.04g KH2PO4, 0.075g MgSO4·7H2O, 0.036g
CaCl2·2H2O, the EDTANa of the citric acid of 0.006g, the ferric citrate of 0.006g, 0.001g2, 0.02g Na2CO3、1mL
A5 liquid microelement, remaining is karst water.
The goat's horn crescent moon algae in logarithmic growth phase is taken, 10% BG-11 for being inoculated in karst water preparation is cultivated by volume
In liquid, carries out illumination constant temperature and expand culture, the intensity of illumination is 4000lux, and the temperature of culture is 25 DEG C, and the time is 7 days.
Frustule, then vacuum freeze drying is collected by centrifugation, obtains the goat's horn crescent moon algae freeze-dried algae powder of karst Aquaponic；
Step 4: the Collection utilization of grease
Ethyl acetate-hexane method is assisted using ultrasonic wave, concrete operations are with step 2, the goat's horn moon that extraction step 3 obtains
Grease in the freeze-dried algae powder of tooth algae, and it is used to prepare biodiesel.
Comparative example 1
Unlike embodiment 1, in the step 3 of this comparative example, karst water is replaced using distilled water, preparation obtains BG-
11 culture solution.Remaining step is identical.
The result shows that:
The scenedesmus frustule of embodiment 2 is 1.1043g, and scenedesmus oil production is 1.1043g × 32.08%=0.3543g.
The scenedesmus frustule of comparative example is 0.3528g, and scenedesmus oil production is 0.3528 × 32.08%=0.1132g.
It can be seen that the oil production of embodiment 1 is 3.13 times of the oil production of comparative example 1.
It can be seen that the richness that the oil-producing microalgae that the present invention is screened using karst region as biomaterial, is generated using karstification
The karst Aquaponic microalgae of the inorganic carbon containing high concentration passes through the Atmospheric CO that algae photosynthesis is fixed by karstification2It is converted into
Biodiesel with economic benefit, to realize remittance important breakthrough of the fundamental research to resource utilization of karst carbon.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of method of karst carbon remittance resource utilization, which comprises the steps of:
Step 1: the screening of karst region microalgae
The microalgae in karst aquifer is collected, then takes former karst water locating for the microalgae, for impregnating the microalgae, saves, obtains in 4 DEG C
To algae stoste；
Using concentration gradient dilution method, algae stoste is isolated and purified, obtains multiple pure strains, and determines its kind respectively；
Step 2: microalgae oil-producing Performance Evaluation
Multiple pure strains that step 1 is obtained are respectively placed in its corresponding culture solution, are carried out illumination constant temperature and are expanded culture,
Frustule, then vacuum freeze drying is collected by centrifugation, obtains the freeze-dried algae powder of multiple pure strains respectively；
Ethyl acetate-hexane method is assisted using ultrasonic wave, extracts the grease in above-mentioned multiple pure strain freeze-dried algae powders, and respectively
The oil productivity for calculating above-mentioned multiple pure strains filters out the highest algae of oil productivity, is named as algae K；
Step 3: the conversion that karst carbon converges
Karst water is taken, is filtered, nutritive salt is added into filtered karst water, preparation obtains the culture solution of algae K；
The algae K for taking the step 2 in logarithmic growth phase to obtain, 10%-15% is inoculated in the culture solution of algae K by volume
In, it carries out illumination constant temperature and expands culture, frustule, then vacuum freeze drying is collected by centrifugation, obtains the freeze-dried algae powder of algae K；
Step 4: the collection of grease
Ethyl acetate-hexane method is assisted using ultrasonic wave, the grease in the freeze-dried algae powder for the algae K that extraction step 3 obtains, and
It is used to prepare biodiesel.
2. a kind of method of karst carbon remittance resource utilization according to claim 1, which is characterized in that described in step 1
The specific method for collecting microalgae in karst aquifer is: in karst aquifer, drawing " ∞ " word with plankton net, or use banister brush
Microalgae is scraped from submerged plant blade face, emergent aquactic plant stem table or stone, then with after planktonic organism net filtration, is transferred to poly- second
In alkene bottle.
3. a kind of method of karst carbon remittance resource utilization according to claim 2, which is characterized in that the planktonic organism
The aperture of net is No. 25.
4. a kind of method of karst carbon remittance resource utilization according to claim 1, which is characterized in that described in step 2
Corresponding culture solution, according to " Chinese Academy of Sciences's fresh water algae library ", network address is http://algae.ihb.ac.cn/
Default.aspx is carried out.
5. a kind of method of karst carbon remittance resource utilization according to claim 1, which is characterized in that described in step 2
The calculation method of oil productivity are as follows: oil productivity=M/N × 100%, in formula, N is the quality (g) of freeze-dried algae powder；M is the quality of grease
(g)。
6. a kind of method of karst carbon remittance resource utilization according to claim 1, which is characterized in that step 2 and step 3
In, the speed of the centrifugation is 10000-12000r/min, and the time of centrifugation is 10min；The temperature of the vacuum freeze drying
It is -55 DEG C, vacuum degree 5Pa, time 12h.
7. a kind of method of karst carbon remittance resource utilization according to claim 1, which is characterized in that described in step 3
The content of inorganic carbon is 3-5mmol/L in karst water；The design parameter of the suction filtration are as follows: the aperture of filter membrane is 0.22 μm, pressure
For 0.098MPa, sucking rate 10L/min；The culture solution of the microalgae kind K is the NaNO containing 1.5g in 1L volume3, 0.04g
KH2PO4, 0.075g MgSO4·7H2O, the CaCl of 0.036g2·2H2O, the ironic citrate of the citric acid of 0.006g, 0.006g
The EDTANa of ammonium, 0.001g2, 0.02g Na2CO3, 1mL A5 liquid microelement, surplus is karst water.
8. a kind of method of karst carbon remittance resource utilization according to claim 7, which is characterized in that the micro member of A5
The group of plain liquid becomes the H of 2.86g/L3BO3, the MnCl of 1.86g/L2·4H2The ZnSO of O, 0.22g/L4·7H2O, 0.39g/L's
Na2MoO4·2H2The CuSO of O, 0.08g/L4·5H2Co (the NO of O and 0.05g/L3)2·6H2O。
9. a kind of method of karst carbon remittance resource utilization according to claim 1, which is characterized in that step 2 and step 4
In, the specific method of the ultrasonic wave auxiliary ethyl acetate-hexane method is: weighing 50mg ± 2mg freeze-dried algae powder respectively, grinds
Broken wall is transferred to centrifuge tube with cover and the mixed liquor of n-hexane and ethyl acetate that 5mL volume ratio is 1:1, sonic oscillation is added
After 15min, 50 DEG C of water-bath 45min vibrate 3 times therebetween, and supernatant is collected in 10000r/min centrifugation, complete primary extract；It will be residual
It stays liquid to repeat extraction 2 times, merges supernatant；The isometric deionized water of combined supernatant is added, and is uniformly mixed,
2000r/min centrifugation, takes upper organic phase, is dried with nitrogen to constant weight.
10. a kind of method of karst carbon remittance resource utilization according to claim 1, which is characterized in that step 2 and step
In 4, the temperature that the illumination constant temperature expands culture is 25 DEG C, and the time is 7 days.
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