CN105148446A - Method for extracting microbial degradation hydrophobic organic compound from cloud point system - Google Patents
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- PLFJWWUZKJKIPZ-UHFFFAOYSA-N 2-[2-[2-(2,6,8-trimethylnonan-4-yloxy)ethoxy]ethoxy]ethanol Chemical compound CC(C)CC(C)CC(CC(C)C)OCCOCCOCCO PLFJWWUZKJKIPZ-UHFFFAOYSA-N 0.000 claims description 23
- IEQAICDLOKRSRL-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-dodecoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO IEQAICDLOKRSRL-UHFFFAOYSA-N 0.000 claims description 18
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 18
- 239000002131 composite material Substances 0.000 claims description 16
- 239000000284 extract Substances 0.000 claims description 12
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 10
- -1 polyoxyethylene Polymers 0.000 claims description 10
- 244000005700 microbiome Species 0.000 claims description 9
- 125000005575 polycyclic aromatic hydrocarbon group Chemical group 0.000 claims description 6
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- YNPNZTXNASCQKK-UHFFFAOYSA-N phenanthrene Chemical compound C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 description 2
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for extracting a microbial degradation hydrophobic organic compound from a cloud point system and belongs to the technical field of environmental protection. The method comprises the following steps that A, a nonionic surfactant is adopted for forming the cloud point system at the temperature lower than the temperature of the microbial degradation hydrophobic organic compound; and B, the cloud point system is used as a medium of the microbial degradation hydrophobic organic compound, and the hydrophobic organic compound in the environment is extracted and solubilized into the degradation culture medium so as to degrade the hydrophobic organic compound. The method is applied to the aspect of degrading hydrophobic organic contaminants of the cloud point system, and no negative effects are caused to microbial survival and degradation vitality; and meanwhile, the bioavailability of the hydrophobic organic compound is improved, and environment pollution treatment is facilitated.
Description
Technical field
The invention belongs to environmental protection technical field, be specifically related to a kind of method extracting microbial degradation hydrophobic organic compound in cloud point system.
Background technology
The restriction that the microbial degradation of hydrophobic organic compound is often subject to that substrate solubility is little, bioavilability is low and growth of microorganism environment is good etc.For this situation, two-phase partitioning system can be used to strengthen organic bioavilability in microbial degradation process, and prior art mainly comprises " the liquid-solid two-phase partitioning system " that " liquid-liquid two-phase partitioning system " and the water of water-organic solvent composition-high molecular polymer forms.
Cloud point system is a kind of two-phase partitioning system be made up of nonionic surfactant water-soluble liquid, is divided into hydrophobic coacervate phase and hydrophilic aqueous phase two parts.The existing a lot of report of the application of cloud point system in microbial conversion hydrophobic compound.Such as, Chinese patent 03142114.8 discloses the application of cloud point system in biology transforms, and Chinese patent 200710046918.X discloses the method for synthesizing L-phenylacetylcarbimolby by microorganism transformation in cloud point system.
Although cloud point system has the application of biological conversion aspect, but cloud point system is as the novel two-phase partitioning system of one, have not been reported in the microbial degradation of hydrophobic organic compound and the bioavilability of pollutant.
Double ring arene and polycyclic aromatic hydrocarbon are common environment hydrophobic organic pollutants, and major part has carcinogenicity, endanger huge.Phenanthrene, as the typical polycyclic aromatic hydrocarbon of one, has carcinogenesis to animal, has spread effect and sensitization to skin.Therefore, the improvement that the biodegradation technique that research and development are modern is used for hydrophobic organic pollutant is significant.
Summary of the invention
For solving prior art Problems existing, the invention provides a kind of method extracting microbial degradation hydrophobic organic compound in cloud point system, achieve the application of cloud point system in hydrophobic organic pollutant degraded, negative effect can not be caused to the survival of microorganism and degraded vigor, improve the bioavilability of hydrophobic organic compound simultaneously, be conducive to the control of environmental pollution.
The invention provides a kind of method extracting microbial degradation hydrophobic organic compound in cloud point system, comprise the steps:
A) non-ionic surface active agent is adopted to form cloud point system at the temperature lower than microbial degradation hydrophobic organic compound;
B) described cloud point system is as the medium of microbial degradation hydrophobic organic compound, by the hydrophobic organic compound extraction solubilising in environment in degraded culture medium, realizes the degraded of hydrophobic organic compound.
Adopt technique scheme, by forming cloud point system, and using the medium of this cloud point system as microbial degradation hydrophobic organic compound, along with the hydrophobic organic compound extraction solubilising in environment is in degraded culture medium, hydrophobic organic compound is fallen by as the nutrition of microorganism and Energy harvesting, thus achieves the degraded of hydrophobic organic compound and the object of pollution control.In the present invention, the key adopting cloud point system to realize microbial degradation hydrophobic organic compound maintains the degraded vigor of microorganism while being to improve organic matter bioavilability.
Preferably, described hydrophobic organic compound is selected from one or more in polycyclic aromatic hydrocarbon and halo polycyclic aromatic hydrocarbon.Polycyclic aromatic hydrocarbon refers to the aromatic hydrocarbons of 2-4 ring.
More preferably, described hydrophobic organic compound is luxuriant and rich with fragrance.
Particularly preferably, described is 30-37 DEG C lower than the organic temperature of microbial degradation.
Preferably, described non-ionic surface active agent is selected from one or more in polyoxyethylene alcohols, octyl phenol polyoxyethylene and polyoxyethylene surfactant.
More preferably, described non-ionic surface active agent is selected from one or more of Triton X-100 (TritonX-100, TritonX-114 and TritonX-45), polyoxyethylene laurel ether (Brij30 and Brij35) and isomeric alcohol polyethenoxy ether (TergitolTMN-3 and TergitolTMN-6).Isomeric alcohol polyethenoxy ether (TergitolTMN-3 and TergitolTMN-6) purchased from American Dow Chemical.
Particularly preferably, described non-ionic surface active agent is selected from polyoxyethylene laurel ether Brij30 and isomeric alcohol polyethenoxy ether TergitolTMN-3 with the (mixture of 4-6) ︰ 5 weight ratio composition.
Preferably, the concentration of described non-ionic surface active agent is 5-120g/L degraded culture medium.
Preferably, described microorganism be Sphingol single-cell (
sphingomonaspolyaromaticivorans).
Preferably, described degraded culture medium is selected from MSM culture medium.
Compared with prior art, the present invention has following advantage: the present invention improves the bioavilability of hydrophobic organic compound by forming cloud point system, achieve the application of cloud point system in hydrophobic organic pollutant degraded, be conducive to the control of environmental pollution.The hydrophobic organic compound being insoluble in water is extracted into coacervate phase by cloud point system, form the organic matter dissolved, because the organic concentration in coacervate phase is higher than organic concentration in aqueous phase, according to biphase equilibrium principle, organic matter is from coacervate phase diffusion mass transfer to aqueous phase, until balance.On the one hand, the organic matter concentration of substrate in water environment residing for microorganism obviously raises, and will significantly improve its biodegradation efficiency; On the other hand, along with in water environment, organic matter is degraded by microorganisms, and the further diffusion mass transfer of the organic matter in cloud point system coacervate phase, to aqueous phase, keeps the balance of substrate in two-phase, thus ensures organic sustaining degradation.The realization key of this process is the bioavilability changing hydrophobic organic compound while cloud point system does not affect microbial degradation vigor.
Accompanying drawing explanation
Fig. 1 is the principle schematic that the present invention extracts microbial degradation.
Detailed description of the invention
The invention will be further described for following instance.The present embodiment is implemented premised on technical solution of the present invention, gives detailed embodiment and process, but protection scope of the present invention is not limited to following embodiment.The experimental technique of unreceipted actual conditions in the following example, usually according to degradation condition or the condition of advising according to manufacturer.
The Sphingol single-cell that the present invention relates to can obtain (deposit number CICC10894) by Chinese industrial Microbiological Culture Collection administrative center.The polycyclic aromatic hydrocarbon of efficient degradation 2-4 ring in the cloud point system that this microbial strains can be formed in the present invention, the present invention extracts the principle schematic of microbial degradation as shown in Figure 1.
Wherein, seed inclined-plane is cultivated as LB solid medium (in 1L water tryptone 10g, dusty yeast 5g, NaCl10g, agar powder 20g) is cultivated 2 days under 30 DEG C of conditions.
Seed activation is cultivated as LB culture medium (in 1L water tryptone 10g, dusty yeast 5g, NaCl10g) is cultivated 2 days in the shaking table of 30 DEG C, 150 revs/min.
Degraded culture medium is inorganic salts MSM culture medium ((NH in 1L water
4)
2sO
41g, Na
2hPO
40.8g, KH
2pO
40.2g, MgSO
47H
2o0.2g, FeCl
33H
2o0.005g, CaCl
22H
2o0.1g, (NH
4)
6mo
7o
244H
2o0.001g), natural pH.Degradation condition is cultivate 5-7 days in the shaking table of 30 DEG C, 150 revs/min.
Luxuriant and rich with fragrance concentration adopts high-performance liquid chromatogram determination.Use HypersilC18 chromatographic column, mobile phase is acetonitrile: water (80:20), and flow velocity is 1mL/min, and determined wavelength is 254nm, and appearance time, at 2.7min, records peak area.Luxuriant and rich with fragrance degraded represents with degradation rate, and its operational formula is: degradation rate=(after luxuriant and rich with fragrance initial concentration-Fei degraded concentration)/luxuriant and rich with fragrance initial concentration * 100%.
In the embodiment of the present invention, non-ionic surface active agent and luxuriant and rich with fragrance addition are in every 100mL degraded culture medium.
embodiment 1
Non-ionic surface active agent TritonX-114 addition be 4g/100mL, under luxuriant and rich with fragrance addition is the condition of 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 69.07%.
embodiment 2
Non-ionic surface active agent TritonX-114 and TritonX-45 with the weight ratio of 8:2 composite and addition be 4g/100mL, under the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 85.41%.
embodiment 3
Non-ionic surface active agent Brij30 addition be 4g/100mL, under luxuriant and rich with fragrance addition is the condition of 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 57.68%.
embodiment 4
Non-ionic surface active agent TergitolTMN-3 addition be 4g/100mL, under luxuriant and rich with fragrance addition is the condition of 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 60.39%.
embodiment 5
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 1g/100mL, under the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 71.31%.
embodiment 6
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 2g/100mL, under the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 87.64%.
embodiment 7
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 3g/100mL, under the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 92.06%.
embodiment 8
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 4g/100mL, under the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 96.53%.
embodiment 9
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 5g/100mL, under the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 91.48%.
embodiment 10
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 6g/100mL, the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 82.40%.
embodiment 11
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 8g/100mL, the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 56.83%.
embodiment 12
Non-ionic surface active agent Brij35 and TergitolTMN-6 with the weight ratio of 5:5 composite and addition be 4g/100mL, the luxuriant and rich with fragrance addition condition that is 40mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 89.01%.
embodiment 13
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 4g/100mL, the luxuriant and rich with fragrance addition condition that is 10mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 48.52%.
embodiment 14
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 4g/100mL, the luxuriant and rich with fragrance addition condition that is 20mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 72.61%.
embodiment 15
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 4g/100mL, the luxuriant and rich with fragrance addition condition that is 80mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 84.99%.
embodiment 16
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 4g/100mL, the luxuriant and rich with fragrance addition condition that is 120mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 79.85%.
embodiment 17
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 4g/100mL, the luxuriant and rich with fragrance addition condition that is 160mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 71.31%.
embodiment 18
Non-ionic surface active agent Brij30 and TergitolTMN-3 with the weight ratio of 5:5 composite and addition be 4g/100mL, the luxuriant and rich with fragrance addition condition that is 200mg/100mL, Sphingol single-cell reacts 6 days by degradation condition, result in the cloud point system formed: luxuriant and rich with fragrance degradation rate is 47.83%.
comparative example
When not adding non-ionic surface active agent and adopting sorbitan monolaurate (Span-20), sorbierite monopalmitate (Span-40) as non-ionic surface active agent, other experiment conditions are identical with embodiment 8, result: luxuriant and rich with fragrance degradation rate is respectively 34.39%, 42.62% and 44.07%.
By contrast embodiment provided by the invention and comparative example known, adopt the method extracting microbial degradation hydrophobic organic compound in cloud point system provided by the invention, luxuriant and rich with fragrance degradation rate can be significantly improved.
Above content is in conjunction with concrete preferred embodiment further description made for the present invention, can not assert that specific embodiment of the invention is confined to these explanations.For general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, some simple deduction or replace can also be made, all should be considered as belonging to protection scope of the present invention.
Claims (10)
1. extract a method for microbial degradation hydrophobic organic compound in cloud point system, it is characterized in that: comprise the steps:
Non-ionic surface active agent is adopted to form cloud point system at the temperature lower than microbial degradation hydrophobic organic compound;
Described cloud point system, as the medium of microbial degradation hydrophobic organic compound, by the hydrophobic organic compound extraction solubilising in environment in degraded culture medium, realizes the degraded of hydrophobic organic compound.
2. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 1, it is characterized in that: described hydrophobic organic compound be selected from polycyclic aromatic hydrocarbon and halo polycyclic aromatic hydrocarbon one or more.
3. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 2, it is characterized in that: described hydrophobic organic compound is for luxuriant and rich with fragrance.
4. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 3, it is characterized in that: described is 30-37 DEG C lower than the organic temperature of microbial degradation.
5. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 1 and 2, it is characterized in that: described non-ionic surface active agent be selected from polyoxyethylene alcohols, octyl phenol polyoxyethylene and polyoxyethylene surfactant one or more.
6. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 5, it is characterized in that: described non-ionic surface active agent be selected from TritonX-100, TritonX-114, TritonX-45, polyoxyethylene laurel ether Brij30, polyoxyethylene laurel ether Brij35, isomeric alcohol polyethenoxy ether TergitolTMN-3 and isomeric alcohol polyethenoxy ether TergitolTMN-6 one or more.
7. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 6, it is characterized in that: described non-ionic surface active agent is selected from Brij30 and TergitolTMN-3 with (the composite mixture obtained of 4-6) ︰ 5 weight ratio.
8. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 1 and 2, it is characterized in that: the concentration of described non-ionic surface active agent is 5-120g/L degraded culture medium.
9. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 1 and 2, it is characterized in that: described microorganism is Sphingol single-cell.
10. extract the method for microbial degradation hydrophobic organic compound in cloud point system according to claim 1 and 2, it is characterized in that: described degraded culture medium is selected from minimal medium.
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| CN105969663A (en) * | 2016-06-21 | 2016-09-28 | 江西理工大学 | Hydrophobic organic compound degrading bacteria screening method based on cloud point system |
| CN105969663B (en) * | 2016-06-21 | 2018-01-09 | 江西理工大学 | A kind of hydrophobic organic compound bacterial isolation method based on cloud point system |
| CN111635065A (en) * | 2020-04-30 | 2020-09-08 | 东莞电建水环境治理技术有限公司 | Ecological bank protection clean system |
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