CN104651346B - A kind of method of the degradation solution and degraded oil component of degraded oil component - Google Patents
A kind of method of the degradation solution and degraded oil component of degraded oil component Download PDFInfo
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- CN104651346B CN104651346B CN201410723019.9A CN201410723019A CN104651346B CN 104651346 B CN104651346 B CN 104651346B CN 201410723019 A CN201410723019 A CN 201410723019A CN 104651346 B CN104651346 B CN 104651346B
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Abstract
It is strengthened function the method and application of microbial degradation petroleum component the present invention relates to a kind of using bamboo charcoal nano, the functional microorganism is Sphingol single-cell (Sphingomonas sp.) GY2B, the petroleum component includes polycyclic aromatic hydrocarbon (phenanthrene) and crude oil, in the case where adding 50-200mg/L bamboo charcoal nano, function stem GY2B is remarkably improved to the degradation capability of petroleum component in water, can be applied to after being immobilized bamboo charcoal nano with functional microorganism GY2B with sodium alginate investment the waste water containing petroleum component biological treatment and petroleum-contaminated water it is biological prosthetic.
Description
Technical field
The invention belongs to the biological treatment of environment organic pollution and technical field of biological remediation, and in particular to a kind of degradation
The method of the degradation solution and degraded oil component of petroleum component.
Background technique
Oil pollution is indispensable important energy source and the raw material of industry in human being's production and life.With petroleum industry
The development of development and harbour petroleum transportation industry, oil spill accident occurrence probability are also increasing.It is announced within 2007 according to maritime office
Data show that China is average and oil spill accident together just occur every three days, oil pollution incident densely occur so frequent, if not
Take quickly, timely respond to, petroleum polluting water body problem will be increasingly prominent, to the littoral and marine eco-environment and human health structure
At seriously threatening.Generally contain PAHs (polycyclic aromatic hydrocarbon), BTEX (four benzene: benzene, toluene, ethylbenzene, diformazan in petroleum and oil product
Benzene) and the noxious materials such as phenols, toxicity is gradually increased by the sequence of alkane, cycloalkane and aromatic hydrocarbon, including polycyclic aromatic hydrocarbon
Many ingredients (such as: benzene, toluene, ethylbenzene, naphthalene, phenanthrene, anthracene, pyrene, fluoranthene, benzo [a] pyrene, benzo [a] anthracene, acenaphthene) inside all by
It is included in Environmental Protection Agency USA (EPA) priority pollutant range.Research and develop the efficient petroleum hydrocarbons removal technology of exploiting economy
It is an urgent demand for improving environmental quality, has a good application prospect.
Biodegrade is one of main eliminating rate of petroleum hydrocarbons in environment.Using in microbial degradation environment
Organic pollutant has been widely accepted, and the advantage is that effect is good, expense is low, secondary pollution is few etc..Currently, people pass through manually
The technologies such as enrichment culture have been isolated to the microorganism that much can degrade or convert certain pollutant.However, this is slightly raw
Object when being added in processing/reparation scene may due to indigenous bacterium harmful competition or be difficult to adapt to environment and lead to often effect
It is bad, therefore the technology for researching and developing a kind of microbial degradation target contaminant that can strengthen function becomes Enviromental Pollution Treatment and repairs
One of the hot subject in multiple field.
Inventor has filtered out the efficient degrading bacteria sphingol of one plant of Phenanthrene early period from contaminated environment
How monad (Sphingomonassp.) GY2B improves plant bacterium GY2B in the actual environment to the degradability of target contaminant
Can, it is the key that be solved required for the present invention.
Summary of the invention
The purpose of the present invention is to overcome the shortcomings of the existing technology with it is insufficient, provide and a kind of strengthened function using bamboo charcoal nano
The method and application of microorganism efficient degradation petroleum component.The present invention is for petroleum component biology drop in above-mentioned oil polluted environment
Problem encountered is solved, a kind of technical method of microbial degradation target contaminant petroleum component that can strengthen function is sought, and
Biological treatment and petroleum-contaminated water by the technology applied to the waste water containing petroleum component is biological prosthetic.
The object of the invention is achieved through the following technical solutions: a kind of degradation solution of degraded oil component, the degradation solution
Including Sphingol single-cell and bamboo charcoal nano, the Sphingol single-cell is Sphingol single-cell (Sphingomonas sp.)
GY2B bacterial strain, the bacterial strain is on 2 24th, 2006 in China typical culture collection center preservation, deposit number are as follows:
CCTCCNo.M206019, the preservation prove 200610034169.4 submission in patent application.
Wherein, the concentration of the bamboo charcoal nano is 50-200mg/L.
Wherein, the Sphingol single-cell is seeded to the initial bacterium amount in degradation solution and reaches 106A/ml or more.
The degradation solution is preferably the inorganic salts culture solution containing Sphingol single-cell and bamboo charcoal nano.
A kind of method of degraded oil component as described above, with sodium alginate fixation by bamboo charcoal nano and the micro- life of function
Object immobilizes, and prepares immobilized microorganism preparation, and the immobilized microorganism preparation is used for the degradation of petroleum component
In.
A kind of method of degraded oil component as described above, the degradation solution include original for the petroleum component of degradation
Oil.
A kind of method of degraded oil component as described above, petroleum component of the degradation solution for degradation includes polycyclic
Aromatic hydrocarbons is luxuriant and rich with fragrance.
Wherein, the concentration of the Phenanthrene is not higher than 100mg/L.
Wherein, the concentration of crude oil is not higher than 500mg/L.
In the above method, preferred operating method are as follows: with sodium alginate fixation by bamboo charcoal nano and functional microorganism into
Row immobilization prepares immobilized microorganism bead, and immobilized spherule is added to the waste water containing petroleum component or the water of oil pollution
In body, the biodegrade for strengthening petroleum component can be realized, shorten the residence time of petroleum component in water.
The present invention has the following advantages and effects with respect to the prior art: bamboo charcoal nano is added to containing polycyclic by the present invention
In the water of aromatic hydrocarbons phenanthrene or crude oil, above-mentioned pollutant is degraded with apparent invigoration effect to functional microorganism GY2B, is shortened above-mentioned
The residence time of pollutant in water.
Detailed description of the invention
Fig. 1: influence of the bamboo charcoal nano to bacterial strain GY2B degrading polycyclic aromatic hydrocarbons phenanthrene.
Fig. 2: bamboo charcoal nano concentration is on the biodegradable influence of Phenanthrene.
Specific embodiment
The present invention is further illustrated below in conjunction with drawings and examples, but embodiments of the present invention are unlimited
Routine techniques progress can refer to for not specifically specified technological parameter in this.
The formula of inorganic salts culture solution is in the present embodiment: 5.0ml/L phosphate buffer (8.5g/L KH2PO4,
21.75g/L K2HPO4·H2O, 33.4g/L Na2HPO4·12H2O, 5.0g/L NH4Cl), 3.0ml/L MgSO4Aqueous solution
(22.5g/L), 1.0ml/L CaCl2Aqueous solution (36.4g/L), 1.0ml/L FeCl3Aqueous solution (0.25g/L), 1.0ml/L is micro-
Secondary element solution (39.9mg/L MnSO4·H2O, 42.8mg/L ZnSO4·H2O, 34.7mg/L (NH4)6Mo7O24·4H2O)。
But it is not intended to limit this technology and uses other culture solutions in application process.
The bamboo charcoal nano used in implementation is purchased from Shanghai Hai Nuo company, for the high temperature bamboo charcoal that Dabie Mountain is fired, uses physical method
Activation, nano level superfine grinding are produced, and are the activated carbon from bamboo powder of 20 nanometers of purity 97%.
The biodegrade of 1 bamboo charcoal nano of embodiment promotion Phenanthrene
Experimental example 1: taking the cyclohexane solution of a certain amount of phenanthrene, and including luxuriant and rich with fragrance quality is 3mg, is placed in the triangular flask of sterilizing,
The inorganic salts culture solution of sterilizing is added after to thiacyclohexane volatilization, adds 6mg bamboo charcoal nano, then according in inoculation refrigerator
The GY2B bacterium solution of preservation, addition inorganic salts culture solution to 30ml are 8 × 10 with bacteria concentration initial in the culture solution postponed6A/
ML, bamboo charcoal nano concentration be 200mg/L, luxuriant and rich with fragrance concentration is 100mg/L, being put into temperature is 30 DEG C, the vibration culture of 150r/min
Case culture 2 days, timing sampling measured luxuriant and rich with fragrance residual quantity, and each experiment sets two parallel laboratory tests, the average value for row experimental result of making even,
As a result as shown in Figure 1.
Comparative example 1: setting is not added bamboo charcoal nano and the control experiment of bacterium is not added, i.e., bamboo charcoal nano and not is not added in culture solution
Add bacterium, remaining is identical as experimental example 1, and timing sampling measures luxuriant and rich with fragrance residual quantity, and each experiment sets two parallel laboratory tests, takes parallel laboratory test
As a result average value, as a result as shown in Figure 1.
Comparative example 2: the control experiment of bamboo charcoal nano is not added in setting, i.e., bamboo charcoal nano is not added in culture solution, remaining and experiment
Example 1 is identical, and timing sampling measures luxuriant and rich with fragrance residual quantity, and each experiment sets two parallel laboratory tests, the average value for row experimental result of making even, knot
Fruit is as shown in Figure 1.
Comparative example 3: the control experiment of bacterium is not added in setting, i.e., bacterium is not added in culture solution, remaining is identical as experimental example 1, timing
Luxuriant and rich with fragrance residual quantity is measured by sampling, each experiment sets two parallel laboratory tests, the average value for row experimental result of making even, as a result as shown in Figure 1.
Luxuriant and rich with fragrance residual quantity uses determined by ultraviolet spectrophotometry, and pre-treatment step is as follows: the bodies such as addition in sample to be tested
Long-pending hexamethylene extracts, mistake anhydrous sodium sulfate after extract liquor concentration, makees reference after constant volume with hexamethylene, with UV1200 spectrophotometric
Meter measures its light absorption value OD at 254nm254。
Bamboo charcoal nano is as shown in Figure 1 to the influence that Sphingol single-cell GY2B degradation is luxuriant and rich with fragrance.As seen from the figure, 200mg/L is added
In experiment after bamboo charcoal nano, the luxuriant and rich with fragrance rate of bacterial strain GY2B degradation is dramatically speeded up: the degradation of 4h phenanthrene is degraded about close to 40%, 8h phenanthrene
80% (and the same period does not add the degradation rate of the sample China and Philippines of bamboo charcoal nano less than 20%), only remain when 18h after 6%, 48h it is luxuriant and rich with fragrance almost
It is degradable.
This example demonstrates that the bamboo charcoal nano of 200mg/L can greatly promote bacterial strain GY2B to luxuriant and rich with fragrance degradation efficiency, shorten luxuriant and rich with fragrance
Residence time in water.
2 bamboo charcoal nano concentration of embodiment is on the biodegradable influence of Phenanthrene
Experimental example 2: the GY2B bacteria suspension after activation is centrifuged, and removes supernatant after centrifugation, retains lower layer's thallus, then
Thallus is washed with phosphate buffer solution, is centrifuged again later, and supernatant is then removed, and lower layer's thallus washs again, in triplicate.
And it is resuspended with minimal medium.Take the above-mentioned bacterium solution of 2mL into 18mL minimal medium, the inorganic salts culture
Added with luxuriant and rich with fragrance and bamboo charcoal nano in base, initial bacteria concentration is 9 × 10 in culture solution after inoculation6A/mL, luxuriant and rich with fragrance concentration be 100mg/L,
The concentration of bamboo charcoal nano is respectively 20,50,200mg/L.It sets simultaneously and the control experiment that bamboo charcoal nano only adds bacterium is not added.Timing sampling
Luxuriant and rich with fragrance residual quantity is measured, each experiment sets two parallel laboratory tests, the average value for row experimental result of making even, as a result as shown in Figure 2.
Experimental example 3: the concentration of the culture solution bamboo charcoal nano after inoculation is 50mg/L, remaining is identical as experimental example 2, periodically takes
Sample measures luxuriant and rich with fragrance residual quantity, and each experiment sets two parallel laboratory tests, the average value for row experimental result of making even, as a result as shown in Figure 2.
Comparative example 4: the concentration of the culture solution bamboo charcoal nano after inoculation is 20mg/L, remaining is identical as experimental example 2, periodically takes
Sample measures luxuriant and rich with fragrance residual quantity, and each experiment sets two parallel laboratory tests, the average value for row experimental result of making even, as a result as shown in Figure 2.
Comparative example 5: the culture solution after inoculation does not add bamboo charcoal nano, remaining is identical as experimental example 2, and timing sampling measurement is luxuriant and rich with fragrance
Residual quantity, each experiment set two parallel laboratory tests, the average value for row experimental result of making even, as a result as shown in Figure 2.
Different bamboo charcoal nano concentration are biodegradable on Phenanthrene to be influenced as shown in Figure 2.As seen from the figure, it is added to and receives
The removal rate of the experimental group phenanthrene of rice bamboo charcoal is significantly faster than that control group, and sees on the whole, and bamboo charcoal nano concentration is higher, luxuriant and rich with fragrance removal
It is faster.The experimental group of 50mg/L, 200mg/L bamboo charcoal nano is added, the removal rate for 24 hours has reached 90%, 48h almost
Removal.Control group without adding bamboo charcoal nano, luxuriant and rich with fragrance removal rate is less than 75% for 24 hours.
This example demonstrates that the bamboo charcoal nano of addition is more, the biodegradable facilitation of Phenanthrene is more obvious,
The facilitation for wherein adding 50-200mg/L bamboo charcoal nano is significant.
The biodegrade of 3 bamboo charcoal nano of embodiment promotion crude oil
Experimental example 4: the GY2B bacteria suspension after taking 1mL to activate is into the inorganic salts culture solution of 10mL, in inorganic salts culture solution
Added with crude oil and bamboo charcoal nano, Crude Oil vibrates volatilization 48h in 30 DEG C, 150r/min constant-temperature shaking incubator in advance,
Initial bacteria concentration is about 7.5 × 10 in culture solution after inoculation6A/mL, crude oil concentration are 500mg/L, the concentration of bamboo charcoal nano is divided
It Wei not 50 and 200mg/L.It sets simultaneously and the control experiment that bamboo charcoal nano only adds bacterium is not added, every group experiment sets two in parallel.30℃,
150r/min cultivates the residual quantity of sampling hexamethylene extraction and determination crude oil after 72h.
The residual quantity of crude oil uses determined by ultraviolet spectrophotometry, makees reference with hexamethylene, with UV2550 spectrophotometer
Its light absorption value OD is measured at wavelength 229nm229, experimental result is shown in Table shown in 1.The results show that with bamboo charcoal nano additive amount
The increase of (0,50,500mg/L), the residual concentration of culture solution Crude Oil gradually decrease, and illustrate that bacterial strain GY2B drops in bamboo charcoal nano
Solving crude oil has apparent facilitation.
Influence of 1 bamboo charcoal nano of table to GY2B degrading crude oil
| Test group | 72h crude oil residual concentration (mg/L) |
| GY2B is not added, bamboo charcoal nano is not added | 425.3 |
| Only add GY2B, bamboo charcoal nano is not added | 416.5 |
| Add GY2B and 50mg/L bamboo charcoal nano | 387.4 |
| Add GY2B and 200mg/L bamboo charcoal nano | 378.6 |
This example demonstrates that the bamboo charcoal nano of addition 50-200mg/L can be obviously promoted degradation of the bacterial strain GY2B to crude oil, contract
The residence time of short crude oil in water.
The compound immobilized microorganism preparation of 4 bamboo charcoal nanos of embodiment-sodium alginate
Sodium alginate investment prepares immobilized microorganism preparation: taking the sodium alginate soln 20mL of 36.5g/L, is added
Ultrasonic mixing uniformly (makes to be granulated bamboo charcoal nano concentration 50mg/L in mother liquor) after 2mg bamboo charcoal nano, then adds 20mL bacterium
The physiological saline bacterium of strain GY2B is outstanding, instills 2% CaCl with syringe after mixing2Molten granulation in liquid, after 1~2h of immobilization
Particle is filtered out with the gauze of cleaning sterilizing, the washing of 0.90% sterile saline, 4 DEG C of preservations, this is immobilization preparation obtained
1.With above-mentioned same method, but it is that 8mg (making to be granulated bamboo charcoal nano concentration 200mg/L in mother liquor) is made admittedly that bamboo charcoal nano, which is added,
Surely change preparation 2.GY2B cell concentration is about 3 × 10 in immobilization preparation 1 or 25A/grain.
The luxuriant and rich with fragrance efficiency of immobilized microorganism preparation degradation: the minimal medium of 10mL phenanthrene containing 100mg/L is taken, is added 30
Immobilized microorganism bead (immobilization preparation 1 or 2) containing bamboo charcoal nano is placed on 30 DEG C, 150r/min constant temperature into culture medium
Culture, the luxuriant and rich with fragrance residual concentration of timing sampling measurement are protected from light in oscillator.The free bacterium that identical bacterium amount is added is set simultaneously as control group.
It the results are shown in Table shown in 2.The result shows that the compound immobilization bacterial strain GY2B of bamboo charcoal nano-sodium alginate is slightly above luxuriant and rich with fragrance degradation rate
Control group free bacterium is to luxuriant and rich with fragrance degradation rate.
2 immobilized microorganism preparation of table and free bacterium are to luxuriant and rich with fragrance degradation effect
This example demonstrates that immobilized microorganism preparation has specific ionization bacterium preferably luxuriant and rich with fragrance degradation property.Immobilized microorganism
In practical applications can less extraneous unfavorable factor influence, the long period can keep bioactivity.In addition, bamboo charcoal nano and bacterium
It is easy to after immobilization and treated that water carries out separation and recycling utilizations, convenient for practical application.
Claims (7)
1. a kind of method for preparing the compound immobilized microorganism preparation of bamboo charcoal nano-sodium alginate, it is characterised in that:
The described method includes: taking the sodium alginate soln 20mL of 36.5g/L, ultrasonic mixing is equal after 2mg or 8mg bamboo charcoal nano is added
It is even, make to be granulated bamboo charcoal nano concentration 50mg/L or 200mg/L in mother liquor, then adds the physiological saline of 20mL bacterial strain GY2B
Bacterium mixes, and instills 2% CaCl with syringe after mixing2Molten granulation in liquid, with the yarn of clean sterilizing after 1~2h of immobilization
Cloth filters out particle, and the immobilized microorganism preparation is made, wherein the immobilization is micro- in the washing of 0.90% sterile saline
The concentration of bacterial strain GY2B is 3 × 10 in biological agent5A/grain, the bacterial strain GY2B is Sphingol single-cell, in 2006 2
The moon 24 is in China typical culture collection center preservation, deposit number are as follows: CCTCC No.M206019.
2. the compound immobilized microorganism preparation of bamboo charcoal nano-sodium alginate being prepared method according to claim 1.
3. a kind of method using the compound immobilized microorganism preparation of bamboo charcoal nano-sodium alginate as claimed in claim 2, special
Sign is: using the microorganism formulation degrading crude oil or Phenanthrene.
4. method as claimed in claim 3, it is characterised in that: the concentration of the Phenanthrene is not higher than 100mg/L.
5. method as claimed in claim 3, it is characterised in that: the concentration of crude oil is not higher than 500mg/L.
6. the compound immobilized microorganism preparation of bamboo charcoal nano-sodium alginate as claimed in claim 2 is containing crude oil or polycyclic aromatic hydrocarbon
Application in the biological treatment of luxuriant and rich with fragrance waste water.
7. the compound immobilized microorganism preparation of bamboo charcoal nano-sodium alginate as claimed in claim 2 is containing crude oil or polycyclic aromatic hydrocarbon
Application during luxuriant and rich with fragrance polluted-water is biological prosthetic.
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| CN108993394A (en) * | 2018-08-17 | 2018-12-14 | 佛山市禅城区诺高环保科技有限公司 | A kind of preparation method of drinking water bamboo charcoal |
| CN109468258A (en) * | 2018-11-15 | 2019-03-15 | 青岛农业大学 | The tolerance propionate bacteria acclimation method in one plant of cud source |
| CN111995066A (en) * | 2020-08-25 | 2020-11-27 | 苏州道源华智环保科技有限公司 | Method for treating petrochemical industry wastewater by adopting biological enhanced microbial inoculum adsorption-embedding mode |
| CN112960778B (en) * | 2021-02-23 | 2022-06-21 | 吴奇桐 | Composition for degrading polycyclic organic matters and preparation method and application thereof |
| CN114107276B (en) * | 2021-11-29 | 2024-03-26 | 青岛大学 | Bacteria stick biochar immobilized phenanthrene degradation microbial agent, and preparation method and application thereof |
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| CN1844361A (en) * | 2006-03-08 | 2006-10-11 | 华南理工大学 | Polycyclic aromatic hydrocarbon degrading bacteria and use thereof |
| CN104059903A (en) * | 2014-06-27 | 2014-09-24 | 浙江工业大学 | Preparation method of efficient solid compound bacterium agent and application of bacterium agent to biological purification of VOCs (volatile organic chemicals) |
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| CN1844361A (en) * | 2006-03-08 | 2006-10-11 | 华南理工大学 | Polycyclic aromatic hydrocarbon degrading bacteria and use thereof |
| CN104059903A (en) * | 2014-06-27 | 2014-09-24 | 浙江工业大学 | Preparation method of efficient solid compound bacterium agent and application of bacterium agent to biological purification of VOCs (volatile organic chemicals) |
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