CN109097284A - A kind of preparation method of the aeroge of the microalgae containing decomposing petroleum hydrocarbon - Google Patents
A kind of preparation method of the aeroge of the microalgae containing decomposing petroleum hydrocarbon Download PDFInfo
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- CN109097284A CN109097284A CN201810884619.1A CN201810884619A CN109097284A CN 109097284 A CN109097284 A CN 109097284A CN 201810884619 A CN201810884619 A CN 201810884619A CN 109097284 A CN109097284 A CN 109097284A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
Abstract
The present invention relates to seawater to administer field, discloses a kind of preparation method and applications of the aeroge of microalgae containing decomposing petroleum hydrocarbon.The present invention selects good biocompatibility, does not have the fixed microalgae of konjaku glucomannan aerosol of toxic action to algae living cells.Also, be added when preparing aerosol high porosity, low-density bombax cotton made of the modified active carbon of acid oxidase, enhance aerosol to the adsorption capacity of oil spilling.Microalgae is fixed in aerosol by the step of this method eliminates first glue, adsorbs again during aerosol formation, and microalgae is not easily to fall off；The contact area of microalgae and petroleum is increased simultaneously, and petroleum is first adsorbed in aeroge, is then degraded by microalgae, improves the degradation efficiency of petroleum hydrocarbon, is conducive to the environment remediation effect for playing microalgae to oil spilling sea area.
Description
Technical field
The present invention relates to the preparation sides that seawater administers the aeroge of field more particularly to a kind of microalgae containing decomposing petroleum hydrocarbon
Method.
Background technique
Along with the increasingly extension of offshore oil exploitation and transport service, blowout, Pipeline Leak and mammoth tanker because
Crash or Oil spills etc. that shipwreck generates caused by oil pollution it is following.Oil spilling is to marine environment, natural resources and supports
Resource etc. is grown with long-term harm, and therefore, oil spill has become the great environmental problem that people must face.Place at present
The common method for managing marine oil spill includes physical method, chemical method and bioanalysis.Physics and chemical method processing marine oil spill exist
There are remaining, secondary pollution problems.Bioanalysis has many advantages, such as that processing method is simple, at low cost, without secondary pollution.However,
Ocean is movement, an opening system, and leading to bioanalysis, there are microorganisms to be easily diluted, and is not easy the problems such as reusing.
Immobilized microorganism technology is that free cell is positioned in the region of restriction, is made using chemically or physically method
Its technology that keeps activity and can recycle.There is this technology microbe density height, cell to be lost less, reaction speed is fast,
The features such as tolerance is strong, product is easily separated.But traditional immobilization adsorption method microorganism adsorption effect is poor, is not easy to be fixed to load
It in body and is easy to run off, causes petroleum hydrocarbon degradation effect poor.
Summary of the invention
In order to solve the above-mentioned technical problems, the present invention provides a kind of preparation sides of the aeroge of microalgae containing decomposing petroleum hydrocarbon
Method.
The specific technical proposal of the invention is: a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon, including with
Lower step:
(1) to microalgae enrichment culture 12~20 days of energy decomposing petroleum hydrocarbon, algae solution is obtained.
(2) algae solution is concentrated, is centrifuged, obtained algae solution precipitating, add water constant volume, shake up to obtain algae suspension.
(3) sterile glycerol solution is added into algae suspension, obtains inoculation liquid, preservation is spare.
(4) by konjaku glucomannan, glycerol, carragheen, hydroxypropyl methyl cellulose, agar, sodium alginate, pectin, deionization
Water is added in beaker.Heating sterilizes after being uniformly mixed, is cooling, obtaining konjaku glucomannan airsetting sol solution.
(5) aseptically, inoculation liquid is injected in konjaku glucomannan airsetting sol solution, is pre-chilled after mixing evenly
Freeze.
(6) the airsetting sol solution crossed through step (5) precooling is put into freeze drier and is freeze-dried, obtained containing microalgae
Konjaku glucomannan aeroge.
The carrier base material of immobilized microalgae of the present invention selects good biocompatibility, does not poison work to algae living cells
Microalgae is fixed in aerosol by konjaku glucomannan aerosol in the preparation process of aerosol.Because aeroge
Aperture is smaller, and using the method that traditional absorption is fixed, inside microalgae aeroge more difficult to get access, and fixed microalgae is easy stream
It moves and falls off.The step of this method eliminates first glue, adsorbs again, it is molten during aerosol formation microalgae to be fixed on gas
In glue, and microalgae is not easily to fall off；The contact area of microalgae and petroleum is increased simultaneously, petroleum is first adsorbed in aeroge, with
It is degraded afterwards by microalgae, improves the degradation efficiency of petroleum hydrocarbon, be conducive to the environment remediation effect for playing microalgae to oil spilling sea area.It is sweet
Oil is frozen solution, improves the survival rate of microalgae during gel-forming.Carragheen, hydroxypropyl methyl cellulose, agar, sea
Synergistic effect is generated between mosanom and konjaku glucomannan, improves the elasticity and antibiotic property of gel, enhances gel effect.Konjaku
Petroleum hydrocarbon is adsorbed in hole by Glucomannan aeroge, then is degraded by microalgae.Under the degradation of microalgae, aeroge
The efficiency of processing seawater oil spilling is remarkably enhanced.
Preferably, in step (4), the acid oxidase for being also 0.1~0.3 part containing parts by weight in the airsetting sol solution
Modified active carbon.Preferably, the active carbon that the acid oxidase is modified is waste and old bombax cotton through carbonization and acid oxidase system
At the modified active carbon of acid oxidase；Preparation method are as follows: waste and old bombax cotton is added in deionized water, in hydro-thermal reaction
Be carbonized at 170~250 DEG C 9~14h in kettle, cooling, is washed with deionized, obtains bombax cotton active carbon, is added into dense
Degree is the concentrated nitric acid of 60~65wt%, and every 1g active carbon 50~100ml concentrated nitric acid handles 2 under conditions of 100~140 DEG C
~8h, cooling, filtering, is in neutrality to filtrate wash with distilled water, and the modified active carbon of acid oxidase is ground to obtain after dry.
The present invention joined when preparing aerosol high porosity, low-density bombax cotton made of acid oxidase it is modified
Active carbon enhances the intensity of the konjaku glucomannan aerosol containing microalgae and its absorption property to oil spilling.Bombax cotton itself
It is the high porosity materials with hollow tubular structure, is good oil absorption material.However, bombax cotton hydrophobicity is strong, by it
When being introduced into the sol solutions of konjaku glucomannan, bombax cotton is difficult to be uniformly mixed with the sol solutions of konjaku glucomannan, is formed
Aeroge in bombax cotton be unevenly distributed, the intensity difference of aeroge.
Active carbon is made in bombax cotton by the present invention, further increases its porosity and oil absorption capacity.Again by active carbon into
Row acid oxidase is modified, its surface is made to contain hydroxyl group.The modified active carbon of acid oxidase is prepared together with konjaku glucomannan
When at aeroge, the hydroxyl of active carbon and the hydroxy combining on konjaku glucomannan surface generate bridging oxygen key.The hole of active carbon itself
Gap and rigidity can increase the pore diameter range and intensity of konjaku glucomannan aeroge, improve the oil absorption efficiency of aeroge.With dense
When nitric acid carries out oxidation modification to active carbon, the hydroxyl of activated carbon surface is controlled by controlling dosage and the modification time of concentrated nitric acid
Base content.The control of the hydroxy radical content of activated carbon surface is emphasis of the invention, and hydroxy radical content is too low, causes active carbon cannot be very
It is good with konjaku glucomannan by bonding together to form stable network structure, cause the intensity of the aeroge of preparation lower；Hydroxyl
When too high levels, the aeroge water imbibition of preparation is strong, causes moisture in aeroge to occupy excessive volume, leads to the suction of aeroge
Oiliness is poor.So needing dosage and the processing time of strict control concentrated nitric acid.
Preferably, the microalgae is chlorella, scenedesmus obliquus, Skeletonema Costatum, tower Ma Alexandria in step (1)
At least one of algae；The culture medium of enrichment culture boils 10~20min with seawater, adds nutritive salt after suction filtration, in culture medium
The concentration of nutritive salt each component is 0.1~0.3g/L of NH4Cl, 0.05~0.1g/L of K2HPO4 and ironic citrate 0.005~
0.02g/L。
To supplement the necessary nutriment of chlorella growth, corresponding nutritive salt, NH are added in the medium4Cl can be mended
Nitrogen charging source, K2HPO4Can supplement the nutrients while supplementing phosphorus source Element Potassium, addition ironic citrate is able to satisfy chlorella and is growing
In the process to the needs of ferro element.
Preferably, the centrifugally operated temperature is 0~4 DEG C in step (2), centrifugation time is 10~20min, from
Scheming revolving speed is 4000~6000r/min；Precipitating is settled to 30~50ml in step (2), obtaining concentration is 7 × 107~7.5 ×
107The algae suspension of cell/mL, water used in constant volume are sterile water.
Preferably, the concentration of the glycerite is 40~60wt% in step (3), glycerite in inoculation liquid
Volumetric concentration is 20~30%, and preservation temperature is 0~4 DEG C.Glycerol is the protection liquid of microalgae, it is ensured that in freezing, microalgae cell
Inside be not in that the ice crystal of bulk bursts microalgae cell, guarantee the survival rate of microalgae.
Preferably, in step (4), the konjaku glucomannan, glycerol, carragheen, hydroxypropyl methyl cellulose, fine jade
Rouge, sodium alginate, pectin, deionized water parts by weight be 0.3~0.5 part of konjaku glucomannan, 2~4 parts of glycerol, carragheen
0.3~0.5 part, 1.4~1.6 parts of hydroxypropyl methyl cellulose, 0.4~0.6 part of agar, 0.5~0.7 part of sodium alginate, pectin
0.5~0.7 part, 100 parts of deionized water；
Preferably, the heating method is heating water bath in step (4), temperature is 70~90 DEG C；Speed of agitator be 600~
800rpm, mixing time are 40~60min；Sterilization method is high pressure steam sterilization, and temperature is 110~120 DEG C, and sterilization time is
10~50min.
Preferably, in step (5), the volume ratio of the inoculation liquid and konjaku glucomannan airsetting sol solution is 0.1~
0.3:1；Precooling temperature is -30~-10 DEG C, and freezing rate is 2~4 DEG C/h, and the precooling time is 8~16h.Preferably, step
Suddenly in (6), the condenser temperature of the freeze-drying is -50~-30 DEG C, and freezing rate is 2~4 DEG C/h, sublimation drying 12
~36h.Ensure the survival rate of maximum microalgae by adjusting the volume ratio of inoculation liquid and konjaku glucomannan airsetting sol solution
With to the highest degradation efficiency of petroleum hydrocarbon.Freezing rate cannot be too fast, and the too fast survival rate that will lead to microalgae of freezing rate reduces,
Under suitable freezing rate and cryogenic temperature, the survival rate of microalgae is 40%~50%.
It is compared with the prior art, the beneficial effects of the present invention are: the carrier base material of immobilized microalgae of the present invention is selected
Good biocompatibility does not have the konjaku glucomannan aerosol of toxic action to algae living cells, in the preparation process of aerosol
It is middle that microalgae is fixed in aerosol.Also, when preparing aerosol be added high porosity, low-density bombax cotton made of
The modified active carbon of acid oxidase enhances the konjaku glucomannan aerosol containing microalgae in the present invention to the absorption property of oil spilling.
Compared with traditional process for fixation, the step of this method eliminates first glue, adsorbs again, and microalgae is not easily to fall off；Increase simultaneously
The contact area of microalgae and petroleum is added, petroleum is first adsorbed in aeroge, is then degraded by microalgae, and petroleum hydrocarbon is improved
Degradation efficiency is conducive to the environment remediation effect for playing microalgae to oil spilling sea area.
Specific embodiment
The present invention will be further described with reference to the examples below.Related device, connection structure in the present invention
And method, if being device well known in the art, connection structure and method without refering in particular to.
Embodiment 1
(1) will be microalgae enrichment culture 20 days of decomposing petroleum hydrocarbon, obtain algae solution.The culture medium of enrichment culture is boiled with seawater
15min adds nutritive salt after suction filtration, and the concentration of nutritive salt each component is NH in culture medium4Cl 0.3g/L, K2HPO4 0.1g/L
With ironic citrate 0.005g/L.
(2) algae solution for obtaining step (1) is concentrated, and under sterile working, 300ml algae solution is distributed into centrifuge tube,
Revolving speed is 5000r/min, temperature is centrifuged 15min under conditions of being 4 DEG C, and supernatant, takes precipitating；Ultrapure water is added into precipitating,
It is settled to 30ml, is shaken up, obtaining concentration is 7.5 × 107The algae suspension of cell/mL.
(3) sterile glycerol solution that concentration is 40wt% is added in the algae suspension for obtaining step (2), obtains the volume of glycerol
The inoculation liquid that concentration is 20%, it is spare in 4 DEG C of preservations.
(4) by 0.3 part of konjaku glucomannan, 2 parts of glycerol, 0.3 part of carragheen, 1.4 parts of hydroxypropyl methyl celluloses, 0.4
Part agar, 0.5 part of sodium alginate, 0.5 part of pectin, 100 parts of deionized waters are added in 500ml beakers.70 DEG C of heating water baths,
It is stirred 40min under the revolving speed of 600rpm, to prevent moisture from evaporating, preservative film is covered on beaker；The konjaku Portugal that will be stirred
Sweet glycan airsetting sol solution is transferred in triangular flask, the high pressure steam sterilization 30min at 115 DEG C, and it is spare to take out cooling.
(5) aseptically, the inoculation liquid injection 100ml konjaku glucomannan airsetting peptization that 10ml contains glycerol is drawn
In liquid, precooling 8h in a low temperature of being put in -10 DEG C after mixing evenly, freezing rate is 2 DEG C/h, promotes ice-nucleus growth.
(6) aeroge that precooling is crossed in step (5) is put into freeze drier, -30 DEG C of condenser temperature, freezing rate is
2 DEG C/h, it is freeze-dried 12h, is dehydrated aeroge completely.
(7) porosity, median pore diameter, the oil absorbency of konjaku glucomannan aeroge etc. in detecting step (6) containing microalgae
Physicochemical property；
(8) the konjaku glucomannan aeroge 0.1g in step (6) containing microalgae is taken, the culture medium containing 0.5g/L petroleum is distributed into
In, 20d is cultivated under the conditions of 25 DEG C；The aeroge being filtered to remove in culture medium detects residual petroleum content.
Embodiment 2
(1) will be microalgae enrichment culture 20 days of decomposing petroleum hydrocarbon, obtain algae solution.The culture medium of enrichment culture is boiled with seawater
15min adds nutritive salt after suction filtration, and the concentration of nutritive salt each component is NH in culture medium4Cl 0.3g/L, K2HPO4 0.1g/L
With ironic citrate 0.02g/L.
(2) algae solution for obtaining step (1) is concentrated, and under sterile working, 300ml algae solution is distributed into centrifuge tube,
Revolving speed is 5000r/min, temperature is centrifuged 15min under conditions of being 4 DEG C, and supernatant, takes precipitating；Ultrapure water is added into precipitating,
It is settled to 40ml, is shaken up, obtaining concentration is 7.3 × 107The algae suspension of cell/mL.
(3) sterile glycerol solution that concentration is 50wt% is added in the algae suspension for obtaining step (2), obtains the volume of glycerol
The inoculation liquid that concentration is 25%, it is spare in 4 DEG C of preservations.
(4) by 0.4 part of konjaku glucomannan, 3 parts of glycerol, 0.4 part of carragheen, 1.5 parts of hydroxypropyl methyl celluloses, 0.2
500ml are added in the modified active carbon of part acid oxidase, 0.5 part of agar, 0.6 part of sodium alginate, 0.6 part of pectin, 100 parts of deionized waters
In beaker.80 DEG C of heating water baths, are stirred 50min under the revolving speed of 700rpm, to prevent moisture from evaporating, cover and protect on beaker
Fresh film；The konjaku glucomannan airsetting sol solution being stirred is transferred in triangular flask, the high pressure steam sterilization at 115 DEG C
It is spare to take out cooling by 30min.
(5) aseptically, the inoculation liquid injection 100ml konjaku glucomannan airsetting peptization that 20ml contains glycerol is drawn
In liquid, precooling 10h in a low temperature of being put in -20 DEG C after mixing evenly, freezing rate is 3 DEG C/h, promotes ice-nucleus growth.
(6) aeroge that precooling is crossed in step (5) is put into freeze drier, -40 DEG C of condenser temperature, freezing rate is
3 DEG C/h, freeze-drying for 24 hours, is dehydrated aeroge completely.
(7) porosity, median pore diameter, the oil absorbency of konjaku glucomannan aeroge etc. in detecting step (6) containing microalgae
Physicochemical property；
(8) the konjaku glucomannan aeroge 0.3g in step (6) containing microalgae is taken, the culture medium containing 0.5g/L petroleum is distributed into
In, 20d is cultivated under the conditions of 25 DEG C；The aeroge being filtered to remove in culture medium detects residual petroleum content.
Wherein, the modified active carbon of acid oxidase is the modified active carbon of acid oxidase made of waste and old bombax cotton；It is made
Preparation Method are as follows: waste and old bombax cotton is added in deionized water, be carbonized at 210 DEG C 12h in hydrothermal reaction kettle, and it is cooling,
It is washed with deionized, obtains bombax cotton active carbon, be added into the concentrated nitric acid that concentration is 65wt%, every 1g active carbon 50ml
Concentrated nitric acid, handles 5h under conditions of 120 DEG C, and cooling, filtering is in neutrality to filtrate wash with distilled water, grinds after dry
The modified active carbon of acid oxidase acid oxidase.
Embodiment 3
(1) will be microalgae enrichment culture 20 days of decomposing petroleum hydrocarbon, obtain algae solution.The culture medium of enrichment culture is boiled with seawater
20min adds nutritive salt after suction filtration, and the concentration of nutritive salt each component is NH in culture medium4Cl 0.2g/L, K2HPO4 0.08g/L
With ironic citrate 0.015g/L.
(2) algae solution for obtaining step (1) is concentrated, and under sterile working, 300ml algae solution is distributed into centrifuge tube,
Revolving speed is 5000r/min, temperature is centrifuged 15min under conditions of being 4 DEG C, and supernatant, takes precipitating；Ultrapure water is added into precipitating,
It is settled to 50ml, is shaken up, obtaining concentration is 7 × 107The algae suspension of cell/mL,.
(3) sterile glycerol solution that concentration is 60wt% is added in the algae suspension for obtaining step (2), obtains the volume of glycerol
The inoculation liquid that concentration is 30%, it is spare in 4 DEG C of preservations.
(4) by 0.5 part of konjaku glucomannan, 4 parts of glycerol, 0.5 part of carragheen, 1.6 parts of hydroxypropyl methyl celluloses, 0.3
500ml are added in the modified active carbon of part acid oxidase, 0.6 part of agar, 0.7 part of sodium alginate, 0.7 part of pectin, 100 parts of deionized waters
In beaker.90 DEG C of heating water baths, are stirred 40min under the revolving speed of 800rpm, to prevent moisture from evaporating, cover and protect on beaker
Fresh film；The konjaku glucomannan airsetting sol solution being stirred is transferred in triangular flask, the high pressure steam sterilization at 115 DEG C
It is spare to take out cooling by 30min.
(5) aseptically, the inoculation liquid injection 100ml konjaku glucomannan airsetting peptization that 30ml contains glycerol is drawn
In liquid, precooling 16h in a low temperature of being put in -30 DEG C after mixing evenly, freezing rate is 4 DEG C/h, promotes ice-nucleus growth.
(6) aeroge that precooling is crossed in step (5) is put into freeze drier, -50 DEG C of condenser temperature, freezing rate is
4 DEG C/h, it is freeze-dried 36h, is dehydrated aeroge completely.
(7) porosity, median pore diameter, the oil absorbency of konjaku glucomannan aeroge etc. in detecting step (6) containing microalgae
Physicochemical property；
(8) the konjaku glucomannan aeroge 0.5g in step (6) containing microalgae is taken, the culture medium containing 0.5g/L petroleum is distributed into
In, 20d is cultivated under the conditions of 25 DEG C；The aeroge being filtered to remove in culture medium detects residual petroleum content.
Wherein, the modified active carbon of acid oxidase is the modified active carbon of acid oxidase made of waste and old bombax cotton；It is made
Preparation Method are as follows: waste and old bombax cotton is added in deionized water, be carbonized at 200 DEG C 14h in hydrothermal reaction kettle, and it is cooling,
It is washed with deionized, obtains bombax cotton active carbon, be added into the concentrated nitric acid that concentration is 65wt%, every 1g active carbon 70ml
Concentrated nitric acid, handles 3h under conditions of 120 DEG C, and cooling, filtering is in neutrality to filtrate wash with distilled water, grinds after dry
The modified active carbon of acid oxidase acid oxidase.
Fuel Oil Remaining in culture medium in embodiment 1-3 is measured using spectrophotometry.It is filtered to remove aeroge, uses 30ml
De- virtue petroleum ether cleans remaining culture medium in culture bottle as solvent, and is transferred to separatory funnel, and anhydrous sodium sulfate is added and inhales
Water removes water layer, draws the upper layer 0.5ml oil, is settled to 10ml with petroleum ether.Light absorption value is measured under the conditions of 420nm, referring to crude oil
Standard curve determines degradation property.Table 1 lists the physicochemical property and oil suction degradability of the aeroge prepared in Examples 1 to 3
The test data of energy.
Table 1
| Performance indicator | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| Porosity | 86.8342% | 90.822% | 92.512% |
| Median pore diameter | 34.2361μm | 31.3468μm | 37.4894μm |
| Tensile strength | 19.8Gpa | 21.7Gpa | 19.4Gpa |
| Microalgae survival rate | 50% | 46.7% | 47.1% |
| Oil absorbency | 14.46g/g | 15.24g/g | 17.95g/g |
| Degradation rate | 56.40% | 60.12% | 58.98% |
As can be seen from Table 1, immobilized microalgae technology of the present invention can effectively fix microalgae, and keep its growth activity, and crude oil is inhaled
Attached effect and microalgae degradation effect are preferable.
The above is only presently preferred embodiments of the present invention, is not intended to limit the invention in any way, it is all according to the present invention
Technical spirit any simple modification, change and equivalent structure transformation to the above embodiments, still fall within skill of the present invention
The protection scope of art scheme.
Claims (10)
1. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon, it is characterised in that the following steps are included:
(1) to microalgae enrichment culture 12 ~ 20 days of energy decomposing petroleum hydrocarbon, algae solution is obtained；
(2) algae solution is concentrated, is centrifuged, obtained algae solution precipitating, add water constant volume, shake up to obtain algae suspension；
(3) sterile glycerite is added into algae suspension, obtains inoculation liquid, preservation is spare；
(4) by konjaku glucomannan, glycerol, carragheen, hydroxypropyl methyl cellulose, agar, sodium alginate, pectin, deionization
Water is added in beaker, heating, sterilizes after being uniformly mixed, is cooling, obtaining konjaku glucomannan airsetting sol solution；
(5) aseptically, inoculation liquid is injected in konjaku glucomannan airsetting sol solution, is pre-chilled after mixing evenly
Freeze；
(6) the airsetting sol solution crossed through step (5) precooling is put into freeze drier and is freeze-dried, obtained containing microalgae
Konjaku glucomannan aeroge.
2. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 1, it is characterised in that: step
Suddenly in (4), active carbon also modified for 0.1 ~ 0.3 part of acid oxidase containing parts by weight in the airsetting sol solution.
3. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 2, it is characterised in that: institute
Stating the modified active carbon of acid oxidase is the waste and old bombax cotton active carbon modified through acid oxidase made of carbonization and acid oxidase；Its
The preparation method comprises the following steps: waste and old bombax cotton is added in deionized water, in hydrothermal reaction kettle at 170 ~ 250 DEG C carbonization 9 ~
14h, it is cooling, it is washed with deionized, obtains bombax cotton active carbon, be added into the concentrated nitric acid that concentration is 60 ~ 65wt%, every 1g
Active carbon 50 ~ 100ml concentrated nitric acid handles 2 ~ 8h, cooling, filtering, wash with distilled water to filter under conditions of 100 ~ 140 DEG C
Liquid is in neutrality, and the modified active carbon of acid oxidase is ground to obtain after dry.
4. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 1, it is characterised in that: step
Suddenly in (1), the microalgae is at least one of chlorella, scenedesmus obliquus, Skeletonema Costatum, Alexandrium tamarense；Enrichment
The culture medium of culture boils 10 ~ 20min with seawater, nutritive salt is added after suction filtration, the concentration of nutritive salt each component is in culture medium
NH4Cl 0.1 ~ 0.3g/L, K2HPO40.005 ~ 0.02g/L of 0.05 ~ 0.1g/L and ironic citrate.
5. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 1, it is characterised in that: step
Suddenly in (2), the centrifugally operated temperature is 0 ~ 4 DEG C, and centrifugation time is 10 ~ 20min, and centrifuge speed is 4000 ~ 6000r/
min；Precipitating is settled to 30 ~ 50ml in step (2), obtaining concentration is 7 × 107~7.5×107The algae suspension of cell/mL, constant volume institute
It is sterile water with water.
6. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 1, it is characterised in that: step
Suddenly in (3), the concentration of the glycerite is 40 ~ 60wt%, and the volumetric concentration of glycerite is 20 ~ 30% in inoculation liquid, preservation
Temperature is 0 ~ 4 DEG C.
7. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 1, it is characterised in that: step
Suddenly in (4), the konjaku glucomannan, glycerol, carragheen, hydroxypropyl methyl cellulose, agar, sodium alginate, pectin, go from
The parts by weight of sub- water are 0.3 ~ 0.5 part of konjaku glucomannan, 2 ~ 4 parts of glycerol, 0.3 ~ 0.5 part of carragheen, hydroxypropyl methyl fibre
Tie up plain 1.4 ~ 1.6 parts, 0.4 ~ 0.6 part of agar, 0.5 ~ 0.7 part of sodium alginate, 0.5 ~ 0.7 part of pectin, 100 parts of deionized water.
8. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 1, it is characterised in that: step
Suddenly in (4), the heating method is heating water bath, and temperature is 70 ~ 90 DEG C；Speed of agitator is 600 ~ 800rpm, and mixing time is
40~60min；Sterilization method is high pressure steam sterilization, and temperature is 110 ~ 120 DEG C, and sterilization time is 10 ~ 50min.
9. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 1, it is characterised in that: step
Suddenly in (5), the volume ratio of the inoculation liquid and konjaku glucomannan airsetting sol solution is 0.1 ~ 0.3:1；Precooling temperature is -30
~ -10 DEG C, freezing rate is 2 ~ 4 DEG C/h, and the precooling time is 8 ~ 16h.
10. a kind of preparation method of the aeroge of microalgae containing decomposing petroleum hydrocarbon according to claim 1, it is characterised in that:
In step (6), the condenser temperature of the freeze-drying is -50 ~ -30 DEG C, and freezing rate is 2 ~ 4 DEG C/h, sublimation drying is 12 ~
36h。
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| CN201810884619.1A CN109097284A (en) | 2018-08-06 | 2018-08-06 | A kind of preparation method of the aeroge of the microalgae containing decomposing petroleum hydrocarbon |
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