CN104276738A - Rapid harmless biological degradation method against waxy oil sludge - Google Patents
Rapid harmless biological degradation method against waxy oil sludge Download PDFInfo
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- CN104276738A CN104276738A CN201410514077.0A CN201410514077A CN104276738A CN 104276738 A CN104276738 A CN 104276738A CN 201410514077 A CN201410514077 A CN 201410514077A CN 104276738 A CN104276738 A CN 104276738A
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- waxy oil
- oil mud
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/10—Nature of the water, waste water, sewage or sludge to be treated from quarries or from mining activities
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/20—Sludge processing
Abstract
The invention provides a rapid harmless biological degradation method against waxy oil sludge. The method is realized by preparing a seed solution, heating and dispersing the waxy oil sludge, preparing a degradation system and performing biological degradation, the method aims at the waxy oil sludge produced in oil production and oil refining processes specially, and substances in the waxy oil sludge can be effectively degraded by using the method. By adopting the method provided by the invention, petroleum hydrocarbon type substances in the oil sludge can be effectively degraded, petroleum hydrocarbons can be partially converted to glycoprotein type substances, and the requirements on harmless treatment of the oil sludge can be finally met. The method utilizes a biological treatment process, prevents secondary pollution and has no influence on an environment; the treatment process is simple, and the operation cost is low; the treatment time is short, and the efficiency is high. A conversion product of the oil sludge can promote the degradation of the oil sludge, which is an unexpected effect obtained in a testing process of the method provided by the invention.
Description
Technical field
The invention belongs to field of solid waste treatment, relate to the process of waxy oil mud, be specifically related to a kind of fast and harmless biodegradation method for waxy oil mud.
Background technology
Noxious waste pollution is harm humans living environment, one of important factor affecting human health, and solid waste is treated as the current difficult problem being difficult to administer.Oily sludge is as a class of solid waste, and a large amount of land occupation, contaminated soil, polluted-water, atmosphere pollution, produce bad impact to the environmental quality of surrounding, is a larger difficult problem in current solid waste disposal.
Disposal of oily sludge is one of important process content of petroleum industry environment protection, and its innoxious and resource treatment technique becomes research emphasis in recent years.In recent years, the process of oily sludge receives much attention, generally adopt landfill both at home and abroad, disperse to execute the methods such as cultivated, concentrated drying and incinerating, chemosetting process, chemical oxidation degraded, chemical solvents cleaning, Coagulation Method solid-liquid separation, there is cost high, easily cause the problem of secondary pollution.Current research person carries out many-sided exploration to oily sludge, and wherein sludge organism process is a kind of development trend.
Biologic treating technique mainly utilizes the oily substance in microbiological deterioration, assimilation oily sludge or the lipid organism in water, makes it be converted into CO
2, H
2a kind of greasy dirt treatment technology of O and other toxicological harmless material.Biologic treating technique is little to environmental influence, and biological treatment is the strengthening of natural degradation process, has the advantages such as treatment effect is good, save energy, less investment, working cost are low, is subject to domestic and international environmental protection industry circle personage common concern and attention at present.At present, existing biologic treating technique is mainly concentrated for recovering the oil, refining oil or the treatment process of contaminated soil and water body in transportation.
Waxy oil mud refers to tank greasy dirt remainder mud after high-pressure steam, pickling, alkali cleaning process clearly.Waxy oil sludge components is complicated, containing a large amount of aged crude oil, wax, bituminous matter, colloid, suspended solid, bacterium, salt, sour gas, corrosion product etc., also comprise a large amount of flocculation agents added in production process, inhibiter, Scale inhibitors, the water conditioners such as sterilant.Therefore, be that current petroleum industry compels a difficult problem to be solved to the process of waxy oil mud.
In prior art, mainly contain solvent extraction, hot water wash, thermolysis, ultrasonic de-oiling technology and biologic treating technique etc. for the common treatment process of waxy oil mud.But the equal Shortcomings of these methods.As large in solvent extraction technology Solvent quantity, cost is high, and solvent recovery process need consume mass energy; Hot water wash technology can produce secondary pollution; Pyrolysis technique energy consumption is high, complicated operation; Current biologic treating technique length consuming time, can not recycle useful resource reclaim.
Summary of the invention
For the deficiency that prior art exists, the object of the invention is to, specially for the waxy oil mud that oil recovery oil refining process produces, a kind of fast and harmless biodegradation method is provided, the hydrocarbon that can effectively degrade in wax greasy dirt, and free from environmental pollution, finally reach the requirement of waxy oil sewage sludge harmlessness process, overcome the problems such as the degraded existed in existing biologic treating technique is slow, resource can not be recycled.
For solving the problems of the technologies described above, the present invention adopts following technical scheme to be achieved:
For a fast and harmless biodegradation method for waxy oil mud, the method with Pseudomonas aeruginosa NY3 and Pseudomonas stutzeri N2 for bacterium source processes waxy oil mud.
Wood chip is added with in described waxy oil mud.
The particle diameter of described wood chip is 0.3mm ~ 1mm, preferred 0.3mm.
Also add in described waxy oil mud and have promotor, described promotor is the mixture of glycerine, magnesium ion or glycerine and magnesium ion.
As above concrete biodegradation method, the method comprises the following steps:
Step one, preparation seed liquor:
Preserve inclined-plane from Pseudomonas aeruginosa NY3 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, obtain Pseudomonas aeruginosa NY3 seed liquor;
Preserve inclined-plane from Pseudomonas stutzeri N2 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, obtain Pseudomonas stutzeri N2 seed liquor;
Step 2, heating dispersion wax greasy dirt:
Waxy oil mud to be degraded is heated to molten state, in waxy oil mud, adds wood chip, dispersed with stirring is even, stops heating, makes the cooling of waxy oil mud, obtain dispersion system;
Step 3, preparation degraded system:
On shaking table, in the dispersion system in step 2, add minimal medium, obtain degraded system;
Step 4, biological degradation:
Add the seed liquor that step one is obtained in the degraded system obtained in step 3, described seed liquor is the mixture of Pseudomonas aeruginosa NY3 seed liquor and Pseudomonas stutzeri N2 seed liquor, on shaking table, at constant temperature aerobic cultivation 5d ~ 8d.
Preferred biodegradation method, the method comprises the following steps:
Step one, preparation seed liquor:
Preserve inclined-plane from Pseudomonas aeruginosa NY3 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, obtain OD
600nmit is the Pseudomonas aeruginosa NY3 seed liquor of 1.82 ± 0.08;
Preserve inclined-plane from Pseudomonas stutzeri N2 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, obtain OD
600nmit is the Pseudomonas stutzeri N2 seed liquor of 1.82 ± 0.08;
Step 2, heating dispersion wax greasy dirt:
Waxy oil mud to be degraded is heated to molten state, in waxy oil mud, adds wood chip, dispersed with stirring is even, stops heating, makes the temperature of waxy oil mud be down to 28 ~ 35 DEG C, obtain dispersion system;
The mass ratio of waxy oil mud and wood chip is (5 ~ 7): 1;
Step 3, preparation degraded system:
On shaking table, in the dispersion system in step 2, add minimal medium, obtain degraded system;
Wherein: in dispersion system, every 2g waxy oil mud correspondence adds 5ml ~ 10ml minimal medium;
Step 4, biological degradation:
Add the seed liquor that step one is obtained in the degraded system obtained in step 3, described seed liquor is the mixture of Pseudomonas aeruginosa NY3 seed liquor and Pseudomonas stutzeri N2 seed liquor, on shaking table, and constant temperature aerobic cultivation 5d ~ 8d at 28 ~ 35 DEG C;
Wherein: in degraded system, every 2g waxy oil mud correspondence adds 5ml ~ 10ml seed liquor;
In described seed liquor, the volume ratio of Pseudomonas aeruginosa NY3 seed liquor and Pseudomonas stutzeri N2 seed liquor is 7:3 ~ 9:1.
Further, also add in the degraded system described in step 3 kind and have promotor, described promotor is the mixture of glycerine, magnesium ion or glycerine and magnesium ion.
Minimal medium described in step 3 comprises (g/L): 5.0g NH
4nO
3, 1mL trace element (2.5g FeSO
47H
2o, 0.1g ZnSO
47H
2o, 0.2g MnCl
24H
2o, 0.024g CoCl
26H
2o, 0.024g NiCl
26H
2o, 0.017g CuCl
22H
2o, 0.109g Na
2moO
42H
2o, 0.062g H
3bO
3, 5mL 12.1M HCl, is dissolved in 1000mL distilled water), 0.1mL 1M MgSO
47H
2o solution, 0.05mL 1M CaCl
22H
2o solution, 100mL phosphate buffered saline buffer, adjust pH to be 7.5, adding distil water is constant volume in 1000mL volumetric flask, 121 DEG C of high-pressure steam sterilizing 30min.
Further, the oil length of described waxy oil mud is 24% ~ 30%.
The present invention compared with prior art, has following technique effect:
Method of the present invention is specially for the waxy oil mud that oil recovery oil refining process produces, the material that can effectively degrade in wax greasy dirt, the hydrocarbon that method of the present invention can be degraded in greasy dirt effectively, part petroleum hydrocarbon is converted into glycoprotein analog material, finally reaches the requirement of greasy dirt harmless treatment.Utilize biological treatment, do not produce secondary pollution, environment is not had an impact; Treatment process is simple, and running cost is low; Treatment time is short, and efficiency is high.
The degraded of greasy dirt converted product to greasy dirt has promoter action, degrade 2 days time, compare the degraded system of not adding greasy dirt converted product, in greasy dirt, the degradation rate of each direct-connected alkane improves between 14-20%, degradation rate that is luxuriant and rich with fragrance and pyrene improves 23% and 20% respectively, and this is a unexpected effect in the application's process of the test.
Accompanying drawing explanation
Fig. 1 is the infrared spectrogram of greasy dirt biological degradation converted product.
Fig. 2 uses different dispersible carrier on the impact of thalline degraded wax greasy dirt efficiency, and wherein, Fig. 2 (a) is wax greasy dirt color atlas of not degrading; The corn cob of Fig. 2 (b) to be dispersible carrier particle diameter be 0.3mm, the color atlas of the residue hydrocarbons after degraded 6d; The wood chip of Fig. 2 (c) to be dispersible carrier particle diameter be 1mm, remains the color atlas of hydrocarbons after degraded 6d.
Fig. 3 is the impact of different wood chip particle diameter on straight chain normal paraffin degradation rate in waxy oil mud.
Fig. 4 is the color atlas remaining hydrocarbons after degraded 8d, and in figure, Fig. 4 (a) is wax greasy dirt color atlas of not degrading; Fig. 4 (b) is waxy oil mud residue hydrocarbons color atlas after degraded 8d.
Below in conjunction with accompanying drawing, explanation is further explained in detail to particular content of the present invention.
Embodiment
Below provide specific embodiments of the invention, it should be noted that the present invention is not limited to following specific embodiment, all equivalents done on technical scheme basis all fall into protection scope of the present invention.
Embodiment 1:
The present embodiment provides a kind of fast and harmless biodegradation method for waxy oil mud, and the method comprises the following steps:
Step one, preparation seed liquor:
The Pseudomonas aeruginosa NY3 of the application is existing bacterial classification, see document: Maiqian Nie, Xihou Yin, Chunyan Ren, Yang Wang, Feng Xu, Qirong Shen.Novel rhamnolipid biosurfactants produced by a polycyclic aromatic hydrocarbon-degrading bacterium Pseudomonas aeruginosa strain NY3.Biotechnology Advances., 2010,28:635 – 643.
The Pseudomonas stutzeri N2 of the application is existing bacterial classification, see document: Fan Yu, Nie Maiqian, Ge Bizhou, just in case, slander quiet, Zhao Jing, Shen little Juan, Chen Xingdou. the screening of resisting high-concentration phenol bacterial strain and degradation characteristic research thereof. environment and safe journal .2012,12 (4): 113-117.
The activation of bacterial classification: by NY3 bacterium seed liquor, N2 bacterium seed liquor is forwarded in extractum carnis solid slant culture base at aseptic operating platform respectively, is placed in 37 DEG C of constant incubators and cultivates 2d, then puts into cold compartment of refrigerator and preserve, and every month at least transfers once.
Extractum carnis solid medium (g/L) is: add 18g ~ 20g agar heated and stirred in 1000ml extractum carnis liquid nutrient medium and be used as inclined-plane and flat board to boiling.
Under aseptic condition, preserve inclined-plane from Pseudomonas aeruginosa NY3 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, 30 DEG C, 160rpm constant temperature oscillation, aerobic cultivation 24h, obtains OD
600nmit is the Pseudomonas aeruginosa NY3 seed liquor of 1.82 ± 0.08.
Under aseptic condition, preserve inclined-plane from Pseudomonas stutzeri N2 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, 30 DEG C, 160rpm constant temperature oscillation, aerobic cultivation 24h, obtains OD
600nmit is the Pseudomonas stutzeri N2 seed liquor of 1.82 ± 0.08.
Extractum carnis liquid nutrient medium (g/L) is: extractum carnis 3g, peptone 10g, sodium-chlor (NaCl) 5g, be dissolved in 1000ml distilled water, be heated to extractum carnis dissolve completely after to during room temperature adjust pH between 7.0 ~ 7.5,121 DEG C of high pressure steam sterilization 30min are for subsequent use.
Step 2, heating dispersion wax greasy dirt:
By 2g oil length be 29.4% waxy oil mud be heated to molten state, oil length and waxy oil account for the weight percent of total waxy oil mud, the wood chip that 0.29g particle diameter is 0.3mm is added in waxy oil mud, the mass ratio of waxy oil mud and wood chip is 7:1, dispersed with stirring is even, stop heating, make the temperature of waxy oil mud be down to 28 ~ 35 DEG C, obtain dispersion system.
Dispersible carrier wood chip is dispersed in the waxy oil mud of melting, for oily substance provides attachment carrier, in process of cooling, waxy oil can evenly be attached to wood chip surface, for Pseudomonas aeruginosa NY3 provides with Pseudomonas stutzeri N2 cell the huge surface-area contacting waxy oil and other organic pollutants, under agitation, attached in the mutual collision process of wood chip of oily substance, oils is resolved gradually, and fully contact with free somatic cells, degraded by cellular metabolism fast and transform, and making cell keep growth fast because of the hydro carbons carbon source of abundance.
Step 3, preparation degraded system:
Dispersion system step 2 obtained adds in 50ml Erlenmeyer flask, on shaking table, in the dispersion system in step 2, add 10ml minimal medium, then add account for whole degraded system volume 12 ‰ mass concentration be 60% promotor glycerine solution, obtain degraded system.
In promotor, magnesium is conducive to the growth of NY3 mycetocyte, promote metabolism and the breeding of bacterium, be conducive to the more azophenlyene compounds of emiocytosis simultaneously, this compounds is electron shuttle body, the redox reaction that cell carries out can be accelerated, especially when cell quantity rises to some amount, when amount of oxygen is not enough in system, as electron acceptor(EA), cell can be maintained and carries out normal redox reaction.Thus promote the degraded of greasy filth.Glycerine is that NY3 mycetocyte produces one of carbon source of rhamnolipid tensio-active agent, additional a small amount of glycerine, and the rhamnolipid produced can improve NY3 bacterium and N2 bacterium to the degradation efficiency of petroleum hydrocarbon.
Minimal medium comprises (g/L): 5.0g NH
4nO
3, 1mL trace element (2.5g FeSO
47H
2o, 0.1g ZnSO
47H
2o, 0.2g MnCl
24H
2o, 0.024g CoCl
26H
2o, 0.024g NiCl
26H
2o, 0.017g CuCl
22H
2o, 0.109g Na
2moO
42H
2o, 0.062g H
3bO
3, 5mL 12.1M HCl, is dissolved in 1000mL distilled water), 0.1mL 1M MgSO
47H
2o solution, 0.05mL 1M CaCl
22H
2o solution, 100mL phosphate buffered saline buffer, adjust pH to be 7.5, adding distil water is constant volume in 1000mL volumetric flask, 121 DEG C of high-pressure steam sterilizing 30min.
Step 4, biological degradation:
Add step one in the degraded system obtained in step 3 and obtain Pseudomonas aeruginosa NY3 seed liquor 9ml, Pseudomonas stutzeri N2 seed liquor 1ml, on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, C14 ~ C18 clearance all reaches more than 85%, C19 ~ C30 clearance between 76% ~ 86%, and C31 ~ C38 clearance reaches between 60% ~ 76%, and clearance that is luxuriant and rich with fragrance and pyrene can reach 34.9% and 34.1% respectively.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.25g converted product, and characterizes it.Result shows, and waxy oil mud converted product is biomacromolecule, and the weight percent that wherein molecular particle size is distributed as 0.37 ~ 20.7 μm accounts for 16.4%; The weight percent of 22.7 ~ 121.8 μm accounts for 72.8%; The weight percent of 133.7 ~ 234.1 μm accounts for 10.8%.
Transform biomacromolecule infrared spectrum as shown in Figure 1, as can be seen from Figure 1,3446cm
-1absorption wide-band is association-NH/-OH stretching vibration absorption peak, and wherein hydroxyl mainly comes from sugared ring, and amino mainly comes from protein.2920cm
-1and 2848cm
-1two weak absorbing peaks, place be methyl or the C-H stretching vibration of methylene radical of sugar.1652,1541,1398cm
-1spectrum peak is from I bands of a spectrum (C=O stretching vibration) of amide group in protein, acid amides II bands of a spectrum (N-H is bending to be superposed with C-N stretching vibration) and acid amides III bands of a spectrum (C-N stretching vibration).1230cm
-1the absorption peak at place may be S=O symmetrical stretching vibration, means in this viscosity conversion product and contains sulfate groups, illustrate in sugar chain it may is sulfated polysaccharides.1080cm in FIG
-1absorption peak is stronger, may be the asymmetric stretch charateristic avsorption band of pyranoid ring, pyranose form glycosidic link or glycoprotein link place C-O-C and C-N-C structure type valence link.973cm
-1place should be the absorption peak of the rocking vibration of the methyne of desoxy sugar.
Can draw through above-mentioned analysis from Fig. 1, containing glycoprotein in converted product product, detect known by sugar and protein content, in 1mg converted product, protein content is 400.62 μ g, and total sugar content is 76.00 μ g.
Comparative example 1: do not add dispersible carrier wood chip
This comparative example is only not add dispersible carrier wood chip with the difference of embodiment 1, and other process is identical with embodiment 1, and on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.The degradation results finally obtained is: in greasy filth, the degradation rate of each direct-connected alkane is 12% ~ 34%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 29.44% and 23.67%.Well below the degradation rate of embodiment 1.
Comparative example 2: change dispersible carrier
This comparative example is only wood chip to be replaced by corn cob with the difference of embodiment 1, and other process is identical with embodiment 1, and on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.As shown in Figure 2, as can be seen from Figure 2, when adopting corn cob to be dispersion system, after 6d, petroleum hydrocarbon only has seldom part to be degraded to degradation results.
Therefore, adopt corn cob to be dispersed in waxy oil mud, after 6d, its degradation efficiency is starkly lower than the degradation rate of wood chip dispersion.
Comparative example 3:
The present embodiment is identical with other process of embodiment 1, and difference is only in step 4, adds 10ml Pseudomonas aeruginosa NY3 seed liquor in degraded system;
Result test characterizes:
In waxy oil mud, the degradation rate of straight chain normal paraffin C14 ~ C34 is all more than 73%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 19.3% and 20.5%.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.15g converted product, the results are shown in Figure 4, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 400.28 μ g, and total sugar content is 66.53 μ g.
Comparative example 4:
The present embodiment is identical with other process of embodiment 1, and difference is only in step 4, adds 10ml Pseudomonas stutzeri N2 seed liquor in degraded system;
Result test characterizes:
In waxy oil mud, the degradation rate of straight chain normal paraffin C14 ~ C34 is all more than 69%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 12.3% and 16.72%.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.14g converted product, the results are shown in Figure 4, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 421.36 μ g, and total sugar content is 70.19 μ g.
Embodiment 2: change wood chip particle diameter
With the difference of embodiment 1, this comparative example is only that the particle diameter of wood chip is had original 0.3mm, be replaced by 0.56mm and 1mm respectively, other process is identical with embodiment 1, and on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.
Degradation results is: straight chain normal paraffin degradation results is shown in Fig. 3, and in petroleum hydrocarbon, the degradation rate of each straight chain normal paraffin obviously reduces when comparatively wood chip particle diameter is 0.3mm in embodiment 1, and degradation rate that is luxuriant and rich with fragrance and pyrene also reduces.
Embodiment 3:
The present embodiment is identical with other process of embodiment 1, difference is only in the biodegradation process of step 4, add step one in the degraded system obtained in step 3 and obtain Pseudomonas aeruginosa NY3 seed liquor 7ml, Pseudomonas stutzeri N2 seed liquor 3ml, on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, C14 ~ C18 clearance all reaches more than 80%, C19 ~ C30 clearance between 70% ~ 83%, and C31 ~ C38 clearance reaches between 53% ~ 66%, and clearance that is luxuriant and rich with fragrance and pyrene can reach 30.9% and 28.1% respectively.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.23g converted product, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 386.77 μ g, and total sugar content is 80.05 μ g.
Embodiment 4:
The present embodiment is identical with other process of embodiment 1, and difference is only in step 2, and in waxy oil mud, add the wood chip that 0.40g particle diameter is 0.3mm, the mass ratio of waxy oil mud and wood chip is 5:1.
In step 4, add step one and obtain Pseudomonas aeruginosa NY3 seed liquor 8ml, Pseudomonas stutzeri N2 seed liquor 2ml in step 3 in the degraded system obtained, on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, the clearance of C14 ~ C31 normal paraffin is between 80% ~ 92.5%, and the normal paraffin clearance of C32 ~ C38 macromolecule is between 66 ~ 71%, and clearance that is luxuriant and rich with fragrance and pyrene can reach 32.5% and 32.1% respectively.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.25g converted product, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 395.62 μ g, and total sugar content is 77.56 μ g.
Embodiment 5:
The present embodiment is identical with other process of embodiment 1, and difference is only in step 2, and in waxy oil mud, add the wood chip that 0.33g particle diameter is 0.3mm, the mass ratio of waxy oil mud and wood chip is 6:1.
In step 3, during preparation degraded system, dispersion system step 2 obtained adds in 50ml Erlenmeyer flask, on shaking table, in the dispersion system in step 2, adds 5ml minimal medium.
In step 4, add step one and obtain Pseudomonas aeruginosa NY3 seed liquor 4ml, Pseudomonas stutzeri N2 seed liquor 1ml in step 3 in the degraded system obtained, on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, the clearance of C14 ~ C31 normal paraffin is between 80% ~ 92.5%, and the normal paraffin clearance of C32 ~ C38 macromolecule is between 66% ~ 71%, and clearance that is luxuriant and rich with fragrance and pyrene can reach 32.5% and 32.1% respectively.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.26g converted product, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 394.51 μ g, and total sugar content is 70.77 μ g.
Embodiment 6:
The present embodiment is identical with other process of embodiment 1, difference is only in step 4, add step one in the degraded system obtained in step 3 and obtain Pseudomonas aeruginosa NY3 seed liquor 4ml, Pseudomonas stutzeri N2 seed liquor 1ml, on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, the degradation rate of normal paraffin C14 ~ C34 is all more than 58%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 24.9% and 26.4%.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.21g converted product, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 400.56 μ g, and total sugar content is 78.89 μ g.
Embodiment 7:
The present embodiment is identical with other process of embodiment 1, difference is only that in step 3, preparation is degraded in plant process, in degraded system, add the promotor Adlerika that 300 μ l volumetric concentrations are 5g/l, then add account for whole degraded system volume 12 ‰ mass concentration be 60% promotor glycerine solution;
In step 4, add step one and obtain Pseudomonas aeruginosa NY3 seed liquor 4ml, Pseudomonas stutzeri N2 seed liquor 1ml in step 3 in the degraded system obtained, on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, the degradation rate of normal paraffin C14 ~ C34 is all more than 88%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 32.9% and 39.4%.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.27g converted product, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 420.31 μ g, and total sugar content is 78.62 μ g.
Embodiment 8:
The present embodiment is identical with other process of embodiment 7, and difference is only that in step 3, preparation is degraded in plant process, only adds the promotor Adlerika that 300 μ l volumetric concentrations are 5g/l in degraded system;
Result test characterizes:
The degradation rate of normal paraffin C14 ~ C34 is all more than 75%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 28.9% and 36.2%.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.28g converted product, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 397.58 μ g, and total sugar content is 74.63 μ g.
Embodiment 9:
The present embodiment is identical with other process of embodiment 1, and difference is only that the oil length of waxy oil mud in step 2 is 24.7%.
The present embodiment is identical with other process of embodiment 1, difference is only that in step 3, preparation is degraded in plant process, in degraded system, add the promotor Adlerika that 300 μ l volumetric concentrations are 5g/l, then add account for whole degraded system volume 12 ‰ mass concentration be 60% promotor glycerine solution;
In step 4, add step one and obtain Pseudomonas aeruginosa NY3 seed liquor 4ml, Pseudomonas stutzeri N2 seed liquor 1ml in step 3 in the degraded system obtained, on shaking table, at 30 DEG C, constant temperature aerobic cultivation 6d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, the degradation rate of straight chain normal paraffin C14 ~ C34 is all more than 83%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 27.5% and 32.6%.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.17g converted product, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 435.68 μ g, and total sugar content is 78.82 μ g.
Embodiment 10:
The present embodiment is identical with other process of embodiment 9, and difference is only in step 4, and on shaking table, at 35 DEG C, constant temperature aerobic cultivation 8d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, the degradation rate of straight chain normal paraffin C14 ~ C34 is all more than 89%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 30.5% and 38.9%.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.24g converted product, the results are shown in Figure 4, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 362.58 μ g, and total sugar content is 66.35 μ g.
Embodiment 11:
The present embodiment is identical with other process of embodiment 9, and difference is only in step 4, and on shaking table, at 28 DEG C, constant temperature aerobic cultivation 5d, completes the degraded to waxy oil mud.
Result test characterizes:
In waxy oil mud, the degradation rate of straight chain normal paraffin C14 ~ C34 is all more than 78%, and degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 25.5% and 27.8%.
Transforming Product yields in waxy oil mud is that every gram of waxy oil mud correspondence produces 0.08g converted product, the results are shown in Figure 4, and it is characterized, characterization result is identical with embodiment 1, prove in converted product product containing glycoprotein, detect known by sugar and protein content, in 1mg waxy oil mud converted product, protein content is 396.28 μ g, and total sugar content is 76.34 μ g.
Embodiment 12:
The present embodiment is identical with other process of embodiment 1, and difference is only: in step 3, degraded system is divided into two kinds, adds 0mg and 50mg greasy dirt converted product respectively; In step 4, on shaking table, at 28 DEG C, constant temperature aerobic cultivation 2d, completes the degraded to waxy oil mud.
Result test characterizes:
Degrade after 2 days, add the degradation rate of straight chain normal paraffin C14 ~ C34 in the degraded system of 0mg greasy dirt converted product all at 50%-57%, degradation rate that is luxuriant and rich with fragrance and pyrene is respectively 59% and 61%; Add the degraded system of 50mg greasy dirt converted product, in greasy dirt, the degradation rate of each direct-connected alkane is compared the degraded system of not adding greasy dirt converted product and is improved between 14%-20%, and degradation rate that is luxuriant and rich with fragrance and pyrene improves 23% and 20% respectively.
Claims (10)
1., for a fast and harmless biodegradation method for waxy oil mud, it is characterized in that: the method with Pseudomonas aeruginosa NY3 and Pseudomonas stutzeri N2 for bacterium source processes waxy oil mud.
2. biodegradation method as claimed in claim 1, is characterized in that: be added with wood chip in described waxy oil mud.
3. biodegradation method as claimed in claim 2, is characterized in that: the particle diameter of described wood chip is 0.3mm ~ 1mm.
4. biodegradation method as claimed in claim 1, is characterized in that: also adding in described waxy oil mud has promotor, and described promotor is the mixture of glycerine, magnesium ion or glycerine and magnesium ion.
5. the biodegradation method as described in claim as arbitrary in Claims 1-4, is characterized in that: the method comprises the following steps:
Step one, preparation seed liquor:
Preserve inclined-plane from Pseudomonas aeruginosa NY3 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, obtain Pseudomonas aeruginosa NY3 seed liquor;
Preserve inclined-plane from Pseudomonas stutzeri N2 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, obtain Pseudomonas stutzeri N2 seed liquor;
Step 2, heating dispersion wax greasy dirt:
Waxy oil mud to be degraded is heated to molten state, in waxy oil mud, adds wood chip, dispersed with stirring is even, stops heating, makes the cooling of waxy oil mud, obtain dispersion system;
Step 3, preparation degraded system:
On shaking table, in the dispersion system in step 2, add minimal medium, obtain degraded system;
Step 4, biological degradation:
Add the seed liquor that step one is obtained in the degraded system obtained in step 3, described seed liquor is the mixture of Pseudomonas aeruginosa NY3 seed liquor and Pseudomonas stutzeri N2 seed liquor, on shaking table, at constant temperature aerobic cultivation 5d ~ 8d.
6. the biodegradation method as described in claim as arbitrary in Claims 1-4, is characterized in that: the method comprises the following steps:
Step one, preparation seed liquor:
Preserve inclined-plane from Pseudomonas aeruginosa NY3 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, obtain OD
600nmit is the Pseudomonas aeruginosa NY3 seed liquor of 1.82 ± 0.08;
Preserve inclined-plane from Pseudomonas stutzeri N2 bacterial strain and cultivate in picking one ring bacterium access beef extract-peptone liquid nutrient medium, obtain OD
600nmit is the Pseudomonas stutzeri N2 seed liquor of 1.82 ± 0.08;
Step 2, heating dispersion wax greasy dirt:
Waxy oil mud to be degraded is heated to molten state, in waxy oil mud, adds wood chip, dispersed with stirring is even, stops heating, makes the temperature of waxy oil mud be down to 28 ~ 35 DEG C, obtain dispersion system;
The mass ratio of waxy oil mud and wood chip is (5 ~ 7): 1;
Step 3, preparation degraded system:
On shaking table, in the dispersion system in step 2, add minimal medium, obtain degraded system;
Wherein: in dispersion system, every 2g waxy oil mud correspondence adds 5ml ~ 10ml minimal medium;
Step 4, biological degradation:
Add the seed liquor that step one is obtained in the degraded system obtained in step 3, described seed liquor is the mixture of Pseudomonas aeruginosa NY3 seed liquor and Pseudomonas stutzeri N2 seed liquor, on shaking table, and constant temperature aerobic cultivation 5d ~ 8d at 28 ~ 35 DEG C;
Wherein: in degraded system, every 2g waxy oil mud correspondence adds 5ml ~ 10ml seed liquor;
In described seed liquor, the volume ratio of Pseudomonas aeruginosa NY3 seed liquor and Pseudomonas stutzeri N2 seed liquor is 7:3 ~ 9:1.
7. the biodegradation method as described in the arbitrary claim of claim 1,2,3 and 4, is characterized in that: the oil length of described waxy oil mud is 24% ~ 30%.
8. biodegradation method as claimed in claim 5, is characterized in that: the oil length of described waxy oil mud is 24% ~ 30%.
9. biodegradation method as claimed in claim 6, is characterized in that: the oil length of described waxy oil mud is 24% ~ 30%.
10. biodegradation method as claimed in claim 5, it is characterized in that: in the degraded system of step 3, add promotor, described promotor is the mixture of glycerine, magnesium ion or glycerine and magnesium ion.
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| CN114717152A (en) * | 2022-04-24 | 2022-07-08 | 北京沃太斯环保科技发展有限公司 | Composite functional microbial inoculum with pseudomonas stutzeri as carrier for in-situ remediation of contaminated soil and application thereof |
| CN114717152B (en) * | 2022-04-24 | 2024-01-09 | 北京沃太斯环保科技发展有限公司 | Composite functional microbial agent taking pseudomonas stutzeri as carrier for in-situ remediation of polluted soil and application thereof |
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