CN106754513A - The modified sodium alginates of TX 100 embed the preparation and application of pseudomonad particle - Google Patents
The modified sodium alginates of TX 100 embed the preparation and application of pseudomonad particle Download PDFInfo
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- CN106754513A CN106754513A CN201611200631.3A CN201611200631A CN106754513A CN 106754513 A CN106754513 A CN 106754513A CN 201611200631 A CN201611200631 A CN 201611200631A CN 106754513 A CN106754513 A CN 106754513A
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- particle
- pseudomonad
- triton
- modified sodium
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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
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- 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
-
- 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
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/32—Hydrocarbons, e.g. oil
- C02F2101/327—Polyaromatic Hydrocarbons [PAH's]
Abstract
Preparation and application process the invention discloses a kind of Triton X 100 (Triton X-100) modified sodium alginate embedding pseudomonad particle.The modified sodium alginates of the TX 100 embedding pseudomonad particle that the present invention is prepared has good removal effect to the organic pollution thing in waste water, and compared to general immobilized microorganism material forming, stability is high, intensity is high, small by external environmental interference, the removal efficiency to Organic Pollutants in Wastewater is significantly increased.
Description
Technical field
The present invention relates to the technical field of immobilized microorganism repairing environment pollution, more particularly to utilize a kind of surface-active
The preparation method and application of organic contamination degradation bacteria embedded particles is strengthened in agent.
Background technology
With developing rapidly for industrial or agricultural, China's environmental pollution is on the rise.Wherein, in terms of soil organic contamination reparation
Focus as current research.On the basis of existing physics, chemistry and bioremediation technology, monotechnics how is overcome not
Foot, is combined and is strengthened, and is the key issue that we are considered to shorten repair time and improve remediation efficiency.It is fixed
It is exactly using a kind of mode of combine d bioremediation to change microbial technique.
Immobilized microorganism technique is that free cell or microorganism are fixed into carrier using method physically or chemically
On, so as to greatly increase the quantity of microorganism, keep its activity and stability and the method that can be recycled.Influence immobilization effect
The key factor of fruit includes process for fixation, fixation support and microorganism selection etc..Immobilized microorganism preparation method master
To include absorption method, investment, cross-linking method and covalent coupling method.Wherein, investment is a kind of the most frequently used method, with operation
Simply, microorganism concn is high, relatively stable advantage is combined with carrier, but microbial cell work may be damaged in embedding process
Property.Therefore, select a kind of with high absorption capacity and small to microbiological effect and be conducive to the material that it is survived solid as embedding
Surely the realization that carrier is beneficial to embedding techniques is changed.
Charcoal be using biomass (such as straw and animal wastes) as raw material anoxic wholly or in part feelings
Through pyrolysis generation rich in carbonaceous material under condition.Because its pore structure is flourishing, the physics and chemistry such as specific surface area is big and ion exchange capacity is high
Property is learned, can be as a kind of efficient adsorbent, while the nutrition such as the microcellular structure on its surface and a large amount of carbon, nitrogen for containing unit
Element, is conducive to perching and breeding for microorganism.Therefore, charcoal is a kind of preferable fixation support.
At present, charcoal immobilized microorganism has been applied in the reparation of polluted-water and soil.Some researchs are
Point out, charcoal can be strengthened to the adsorption capacity of organic matter and the degradation property of microorganism by some chemical regulators.But
So far, do not study also by adding the method for surfactant in microorganism embedding system come while strengthening embedding
The absorption property of grain and the degrading activity of microorganism.
The content of the invention
It is an object of the invention to prepare Trion X-100 (Triton X-100) enhanced biological using investment
Charcoal immobilization organic pollutant degradation bacterium particle removes organic pollutants.Because Trion X-100 are to biological carbon surface
The modified and influence to aspects such as microbial cell hydrophobicitys, had both enhanced suction-operated and charcoal of the charcoal to pollutant
The strength of stability of immobilization organic pollutant degradation bacterium, promotes degradation of the degradation bacteria to pollutant, so that on the whole again
Improve removal efficiency of the charcoal immobilization organic pollutant degradation bacterium particle to organic pollutants.
The invention provides the preparation method that a kind of TX-100 modified sodium alginates embed pseudomonad particle, step is such as
Under：
Step 1) a kind of pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) is incubated at containing the inorganic of acenaphthene
In salt culture medium, resuspension is centrifuged after 30-40 hours；
Step 2) make OD using minimal medium regulation bacteria suspension600=0.2~1, add Triton X-100, order
Triton X-100 concentration is 50mg/L~200mg/L；
Step 3) by charcoal with containing Triton X-100 P. aeruginosa bacteria suspension in mass ratio 1:100 mixing, stir
After mixing, then by mixed liquor and sodium alginate soln by volume 2：1 mixing；
Step 4) obtained mixed liquor is dropwise added drop-wise in calcium chloride solution, obtain the embedding of TX-100 modified sodium alginates
Pseudomonad particle, is stored at room temperature solidification.
Pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) in the present invention protected on December 28th, 2015
Ensconce China typical culture collection center, preserving number CCTCC NO：M2015786, preservation address is that Wuhan, China Wuhan is big
Learn.
Further, the step 3) in charcoal system is cracked under 500 DEG C of anoxia conditions for rice straw, bamboo powder, rice bran
.
Further, the step 3) in sodium alginate soln concentration be 3.5%.
Further, wherein comprising the following steps that：
Step 1) will be incubated under the conditions of a kind of 30 DEG C of pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ)
In minimal medium containing 3.5mg/L acenaphthenes, resuspension is centrifuged after 36 hours；
Step 2) make OD using minimal medium regulation bacteria suspension600=0.2~1, add Triton X-100, order
Triton X-100 concentration is 50mg/L~200mg/L；
Step 3) by charcoal with containing Triton X-100 P. aeruginosa bacteria suspension in mass ratio 1:100 mixing, stir
After mixing absorption 30min, it is transferred in 3.5% sodium alginate soln and shakes up, mixed liquor and sodium alginate soln by volume 2：1
Mixing；
Step 4) above-mentioned mixed liquor is dropwise added drop-wise in 2% calcium chloride solution, obtain the modified alginic acids of TX-100
Sodium embeds pseudomonad particle, after solidification 12h is stood under the conditions of 25 DEG C of room temperature, leaches particle and with ultrapure water 3 times.
The present invention also provides a kind of TX-100 modified sodium alginates embedding pseudomonad particle, and the particle is by following system
Preparation Method is prepared：
Step 1) a kind of pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) is incubated at containing the inorganic of acenaphthene
In salt culture medium, resuspension is centrifuged after 30-40 hours；
Step 2) make OD using minimal medium regulation bacteria suspension600=0.2~1, add Triton X-100, order
Triton X-100 concentration is 50mg/L~200mg/L；
Step 3) by charcoal with containing Triton X-100 P. aeruginosa bacteria suspension in mass ratio 1:100 mixing, stir
After mixing, then by mixed liquor and sodium alginate soln by volume 2：1 mixing；
Step 4) obtained mixed liquor is dropwise added drop-wise in calcium chloride solution, obtain the embedding of TX-100 modified sodium alginates
Pseudomonad particle, is stored at room temperature solidification.
Further, the step 1) in pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) be preserved in China
Type Tissue Collection, preserving number CCTCC NO：M2015786.
Further, the step 3) in charcoal system is cracked under 500 DEG C of anoxia conditions for rice straw, bamboo powder, rice bran
.
The present invention also provides a kind of above-mentioned TX-100 modified sodium alginates embedding pseudomonad particle in waste water is removed
The application of acenaphthene.
Further, TX-100 modified sodium alginates embedding pseudomonad particle is direct plungeed into the ratio of 10g/L organic
In waste water, after mixing 6h, the acenaphthene in waste water is can remove.
The present invention is in fully research TrionX-100 to organic pollutant degradation bacterium performance, TrionX-100 and embedding medium sea
On the basis of mosanom reciprocation, TrionX-100 concentration and embedding system matching, Triton X-100 reinforcings have been invented
Organic contamination degradation bacteria embedded particles, and this novel biomaterial is applied in the treatment of organic polluting water, achieve
Good treatment effect.The preparation method of charcoal immobilization organic pollutant degradation bacterium particle of the invention is simple, obtains
TX-100 modified sodium alginates embed pseudomonad particle compared to general immobilized microorganism material forming, and stability is high,
Intensity is high, small by external environmental interference, recyclable and reusing, is also more suitably applied in reactor；To having in waste water
The removal efficiency of organic pollutants is significantly increased.
Specific embodiment
The present invention is described in further detail with reference to embodiment：
Embodiment 1
Pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) is incubated at containing 3.5mg/L under the conditions of 30 DEG C
In the minimal medium of acenaphthene, it is centrifuged after 36 hours, resuspension.Bacteria suspension is adjusted with minimal medium, OD is obtained600It is 0.2
Bacterium solution 20mL.The 5g/L Triton X-100 solution of 0.4mL is added in above-mentioned bacterium solution, and (Triton X-100 concentration is
100mg/L).By (mass ratio 1 in the 0.2g charcoals above-mentioned 20mL bacterium solutions containing Triton X-100 of addition:100), stirring and adsorbing
30min.Above-mentioned mixed liquor is added to after being mixed in the sodium alginate soln of 10ml 3.5%, is added dropwise to 1000 μ l liquid-transfering guns
In 2% calcium chloride solution, Triton X-100 enhanced biological charcoal immobilization organic pollutant degradation bacterium particles, room temperature 25 are obtained
After solidification 12h is stood under the conditions of DEG C, with ultrapure water 3 times, suction filtration is dried.
The charcoal immobilization organic pollutant degradation bacterium particle that will be prepared all puts into 200mL 3.5mg/L containing acenaphthene
Organic wastewater in mix 3d, the clearance to acenaphthene can reach 89.6%.As control, Triton X-100 reinforcings are not added with
Under the conditions ofThe charcoal immobilization degradation bacteria particle for preparing is embedded with sodium alginate, to the clearance of acenaphthene under similarity condition
70.0%.It can be seen that, after being strengthened using Triton X-100, immobilization organic pollutant degradation bacterium particle is improved to the clearance of acenaphthene
28%.
Embodiment 2
Pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) is incubated at containing 3.5mg/L under the conditions of 30 DEG C
In the minimal medium of acenaphthene, it is centrifuged after 36 hours, resuspension.Bacteria suspension is adjusted with minimal medium, OD is obtained600It is 1
Bacterium solution 20mL.The 5g/L Triton X-100 solution of 0.4mL is added in above-mentioned bacterium solution, and (Triton X-100 concentration is
100mg/L).By (mass ratio 1 in the 0.2g charcoals above-mentioned 20mL bacterium solutions containing Triton X-100 of addition:100), stirring and adsorbing
30min.Above-mentioned mixed liquor is added to after being mixed in the sodium alginate soln of 10ml 3.5%, is added dropwise to 1000 μ l liquid-transfering guns
In 2% calcium chloride solution, Triton X-100 enhanced biological charcoal immobilization organic pollutant degradation bacterium particles, room temperature 25 are obtained
After solidification 12h is stood under the conditions of DEG C, with ultrapure water 3 times, suction filtration is dried.
All input 200L contains acenaphthene 3.5mg/L's to the charcoal immobilization organic pollutant degradation bacterium particle that will be prepared
48h is mixed in organic wastewater, the clearance to acenaphthene can reach 93.8%.As control, Triton X-100 reinforcings are not added with
Under the conditions of with sodium alginate embed prepare charcoal immobilization degradation bacteria particle, to the clearance of acenaphthene under similarity condition
72.5%.It can be seen that, after being strengthened using Triton X-100, immobilization organic pollutant degradation bacterium particle is improved to the clearance of acenaphthene
29.3%.
Embodiment 3
Pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) is incubated at containing 3.5mg/L under the conditions of 30 DEG C
In the minimal medium of acenaphthene, it is centrifuged after 36 hours, resuspension.Bacteria suspension is adjusted with minimal medium, OD is obtained600It is 1
Bacterium solution 20mL.The 5g/L Triton X-100 solution of 0.8mL is added in above-mentioned bacterium solution, and (Triton X-100 concentration is
200mg/L).By (mass ratio 1 in the 0.2g charcoals above-mentioned 20mL bacterium solutions containing Triton X-100 of addition:100), stirring and adsorbing
30min.Above-mentioned mixed liquor is added to after being mixed in the sodium alginate soln of 10ml 3.5%, is added dropwise to 1000 μ l liquid-transfering guns
In 2% calcium chloride solution, Triton X-100 enhanced biological charcoal immobilization organic pollutant degradation bacterium particles, room temperature 25 are obtained
After solidification 12h is stood under the conditions of DEG C, with ultrapure water 3 times, suction filtration is dried.
The charcoal immobilization organic pollutant degradation bacterium particle that will be prepared all puts into 200mL 3.5mg/L containing acenaphthene
Organic wastewater in mix 6h, the clearance to acenaphthene can reach 91.1%.As control, Triton X-100 reinforcings are not added with
Under the conditions of with sodium alginate embed prepare charcoal immobilization degradation bacteria particle, to the clearance of acenaphthene under similarity condition
52.4%.It can be seen that, after being strengthened using Triton X-100, immobilization organic pollutant degradation bacterium particle is improved to the clearance of acenaphthene
73.9%.
In a word, presently preferred embodiments of the present invention, all equalizations made according to scope of the present invention patent be the foregoing is only
Change and modification, should all belong to the covering scope of patent of the present invention.
Claims (10)
1.TX-100 modified sodium alginates embed the preparation method of pseudomonad particle, it is characterised in that step is as follows：
Step 1) a kind of pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) is incubated at the inorganic salts training containing acenaphthene
Support in base, resuspension is centrifuged after 30-40 hours；
Step 2) make OD using minimal medium regulation bacteria suspension600=0.2~1, Triton X-100 are added, make Triton
X-100 concentration is 50mg/L~200mg/L；
Step 3) by charcoal with containing Triton X-100 P. aeruginosa bacteria suspension in mass ratio 1:100 mixing, after stirring,
Again by mixed liquor and sodium alginate soln by volume 2：1 mixing；
Step 4) obtained mixed liquor is dropwise added drop-wise in calcium chloride solution, obtain the embedding of TX-100 modified sodium alginates false single
Born of the same parents' bacterium particle, is stored at room temperature solidification.
2. TX-100 modified sodium alginates according to claim 1 embed the preparation method of pseudomonad particle, its feature
It is：The step 1) in pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) be preserved in Chinese Typical Representative culture
Collection, preserving number CCTCC NO：M2015786.
3. TX-100 modified sodium alginates according to claim 1 embed the preparation method of pseudomonad particle, its feature
It is：The step 3) in charcoal it is obtained for rice straw, bamboo powder, rice bran are cracked under 500 DEG C of anoxia conditions.
4. TX-100 modified sodium alginates according to claim 1 embed the preparation method of pseudomonad particle, its feature
It is：The step 3) in sodium alginate soln concentration be 3.5%.
5. TX-100 modified sodium alginates according to claim 1 embed the preparation method of pseudomonad particle, its feature
It is：Step is as follows：
Step 1) will be incubated under the conditions of a kind of 30 DEG C of pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) and contain
In the minimal medium of 3.5mg/L acenaphthenes, resuspension is centrifuged after 36 hours；
Step 2) make OD using minimal medium regulation bacteria suspension600=0.2~1, Triton X-100 are added, make Triton
X-100 concentration is 50mg/L~200mg/L；
Step 3) by charcoal with containing Triton X-100 P. aeruginosa bacteria suspension in mass ratio 1:100 mixing, stirring is inhaled
After attached 30min, it is transferred in 3.5% sodium alginate soln and shakes up, mixed liquor and sodium alginate soln by volume 2：1 mixes
Close；
Step 4) above-mentioned mixed liquor is dropwise added drop-wise in 2% calcium chloride solution, obtain the modified sodium alginate bags of TX-100
Pseudomonad particle is buried, after solidification 12h is stood under the conditions of 25 DEG C of room temperature, particle is leached and with ultrapure water 3 times.
6. a kind of TX-100 modified sodium alginates embed pseudomonad particle, it is characterised in that the particle is by following preparation
Method is prepared：
Step 1) a kind of pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) is incubated at the inorganic salts training containing acenaphthene
Support in base, resuspension is centrifuged after 30-40 hours；
Step 2) make OD using minimal medium regulation bacteria suspension600=0.2~1, Triton X-100 are added, make Triton
X-100 concentration is 50mg/L~200mg/L；
Step 3) by charcoal with containing Triton X-100 P. aeruginosa bacteria suspension in mass ratio 1:100 mixing, after stirring,
Again by mixed liquor and sodium alginate soln by volume 2：1 mixing；
Step 4) obtained mixed liquor is dropwise added drop-wise in calcium chloride solution, obtain the embedding of TX-100 modified sodium alginates false single
Born of the same parents' bacterium particle, is stored at room temperature solidification.
7. TX-100 modified sodium alginates according to claim 6 embed pseudomonad particle, it is characterised in that the step
It is rapid 1) in pseudomonas aeruginosa (Pseudomonas aeruginosa JXQ) be preserved in China typical culture collection center, protect
Tibetan CCTCC NO：M2015786.
8. TX-100 modified sodium alginates according to claim 6 embed pseudomonad particle, it is characterised in that：The step
It is rapid 3) in charcoal it is obtained for rice straw, bamboo powder, rice bran are cracked under 500 DEG C of anoxia conditions.
9. a kind of TX-100 modified sodium alginates as claimed in claim 6 embed pseudomonad particle acenaphthene in waste water is removed
Using.
10. application according to claim 9, it is characterised in that TX-100 modified sodium alginates are embedded into pseudomonad
Grain is direct plungeed into organic wastewater in the ratio of 10g/L, after mixing 6h, can remove the acenaphthene in waste water.
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| CN201611200631.3A CN106754513B (en) | 2016-12-22 | 2016-12-22 | Preparation and application of TX-100 modified sodium alginate embedded pseudomonas particles |
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|---|---|---|---|
| CN201611200631.3A CN106754513B (en) | 2016-12-22 | 2016-12-22 | Preparation and application of TX-100 modified sodium alginate embedded pseudomonas particles |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107475237A (en) * | 2017-09-29 | 2017-12-15 | 长沙理工大学 | A kind of immobilization pseudomonas aeruginosa biological adsorption agent and preparation method thereof |
| CN108085312A (en) * | 2017-12-08 | 2018-05-29 | 沈阳化工大学 | A kind of preparation method of environment-friendly type organic wastewater biological cleanser |
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| CN101818140A (en) * | 2010-04-28 | 2010-09-01 | 湖南大学 | Immobilization pseudomonas aeruginosa as well as preparation method and application thereof |
| CN103304040A (en) * | 2013-05-23 | 2013-09-18 | 东北电力大学 | Method for treating nitrogen-containing waste water by utilizing carbon fiber felt-immobilized pseudomonas aeruginosa |
| CN103820370A (en) * | 2014-03-07 | 2014-05-28 | 江苏农林职业技术学院 | Pseudomanas aeruginosa and application thereof |
| CN105695443A (en) * | 2016-03-09 | 2016-06-22 | 浙江工商大学 | Triton X-100-enhanced biochar immobilized microbial material as well as preparation method and application of biochar immobilized microbial material |
-
2016
- 2016-12-22 CN CN201611200631.3A patent/CN106754513B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101818140A (en) * | 2010-04-28 | 2010-09-01 | 湖南大学 | Immobilization pseudomonas aeruginosa as well as preparation method and application thereof |
| CN103304040A (en) * | 2013-05-23 | 2013-09-18 | 东北电力大学 | Method for treating nitrogen-containing waste water by utilizing carbon fiber felt-immobilized pseudomonas aeruginosa |
| CN103820370A (en) * | 2014-03-07 | 2014-05-28 | 江苏农林职业技术学院 | Pseudomanas aeruginosa and application thereof |
| CN105695443A (en) * | 2016-03-09 | 2016-06-22 | 浙江工商大学 | Triton X-100-enhanced biochar immobilized microbial material as well as preparation method and application of biochar immobilized microbial material |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107475237A (en) * | 2017-09-29 | 2017-12-15 | 长沙理工大学 | A kind of immobilization pseudomonas aeruginosa biological adsorption agent and preparation method thereof |
| CN108085312A (en) * | 2017-12-08 | 2018-05-29 | 沈阳化工大学 | A kind of preparation method of environment-friendly type organic wastewater biological cleanser |
| CN108085312B (en) * | 2017-12-08 | 2021-08-06 | 沈阳化工大学 | Preparation method of environment-friendly organic wastewater biological purifying agent |
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| CN106754513B (en) | 2020-08-14 |
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