CN108048374A - A kind of degradation bacteria strains JN4 of oily sludge petrochina hydro carbons and its application - Google Patents
A kind of degradation bacteria strains JN4 of oily sludge petrochina hydro carbons and its application Download PDFInfo
- Publication number
- CN108048374A CN108048374A CN201810115860.8A CN201810115860A CN108048374A CN 108048374 A CN108048374 A CN 108048374A CN 201810115860 A CN201810115860 A CN 201810115860A CN 108048374 A CN108048374 A CN 108048374A
- Authority
- CN
- China
- Prior art keywords
- degradation
- oily sludge
- bacterial strain
- pseudomonas stutzeri
- repaired
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/38—Pseudomonas
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Water Supply & Treatment (AREA)
- Environmental & Geological Engineering (AREA)
- Hydrology & Water Resources (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The present invention discloses efficient degrading bacteria JN4 and its application of one plant of oily sludge petrochina hydro carbons, belongs to technical field of biological remediation.The degradation bacteria JN4 is preserved in China General Microbiological culture presevation administrative center, and preserving number is CGMCC NO.14975, and classification number belongs to Pseudomonas stutzeri for Pseudomonas stutzeri.Bacterial strain JN4 purposes of the present invention with petroleum hydrocarbon degradation function is that the improvement of oily sludge is carried out in a manner of degraded oil hydro carbons.The bacterium was 71.05% to the degradation rate of total petroleum hydrocarbon in oily sludge (TPH) in 30 days.Single bacterial strain provided by the invention is to n C10~n C30With apparent degradation capability, by GC MS analysis shows, the bacterial strain is stronger to the degradation capability of cycloalkane naphthalane, indenes alkane, d-homoandrostane.The bacterium is 25 40 DEG C, pH value 5 11 to the most suitable degradation condition of petroleum pollution.The bioanalysis is used for the recovery technique of oily sludge, has efficient, at low cost, advantages of environment protection.
Description
Technical field
The present invention relates to technical field of biological remediation, and in particular to bacterial strain that the degradation of one plant of oily sludge is repaired and its
Application in degradation oily sludge.
Background technology
Since 20th century, oil is had become as one of most important energy of the mankind, and same what is largely exploited and used
When, oil and its processed goods are endangered caused by environment and the mankind and are also on the rise.China's crude oil since reform and opening-up is produced per year
For amount more than 100,000,000 t, the China for becoming one of ten big oil-producing country of the world possesses more than 400 exploration, the oil field of exploitation and oil gas field,
National 25 provinces (city) and autonomous region are distributed in the ratio for accounting for national territorial area 3%.However economic technology fast development with
And during oil demand amount constantly rises, we still fall behind the technical matters that oil field region is recovered the oil relatively, ring
Border influence appraisement system is relatively not perfect, while the recovery technique of Petroleum concentration is relatively lacked so that China's oil class pollutes
Degree is far above developed country, and aggravates situation in accumulation year by year.
Greasy filth was the main waste pollution problem of each elephant generally existing of China, early in State Environmental Protection Administration in 1998
Greasy filth is just classified as hazardous waste (waste classification HW08), it is desirable that harmless treatment must be carried out to oily sludge the like waste.
Since the source of oily sludge is different, component, property are also not quite similar.By the sampling analysis to all kinds of oily sludges, contain
The general oil-containing 15%~50% of greasy dirt.It is larger to environmental hazard.According to《People's Republic of China's promotion law on cleaner production》It will
Harmless treatment must be carried out to oily sludge by asking.It is effective a large amount of human and material resources progress oily sludges have been employed both at home and abroad at present
The research of processing, attempted in succession cure landfill, burn, elution, chemical oxidation and it is biological prosthetic the methods of handle oily sludge.
Bioremediation technology have advantage of lower cost, it is safe, easy to operate, do not easily cause secondary pollution, can implement in situ at
The advantages that reason, it is considered to be most promising reparation means.But since oily sludge complicated component is difficult to degrade, at present both at home and abroad
It is relatively fewer to the microbial degradation repairing research of oily sludge.
CN1594549A discloses a kind of preparation of pseudomonas stutzeri strain and its to organic compounds containing sulfur desulfurization
In application technology, the bacterial strain be Pseudomonas stutzeri UP-1；It is dirty that it is mainly derived from the higher oil field of sulfur-bearing
In water, oil field sludge and the soil that is polluted by the oil, which can be by specifically cultivating, separating and preparation means obtain, it
Liquid medium body cell, resting cell and immobilized cell can organic compounds containing sulfur be acted on by Kodama routes,
Generation is dissolved in the sulfurous organic compound of water.
CN101074425A discloses a nitro-reductive pseudomonads and its application.The microorganism of the present invention is from oil
The Pseudomonas nitroreducens Pseudomonas that separation and purified culture obtain in the activated sludge of chemical plant sewage treatment plant
Nitroreducens S1 have benzothiazole efficient single-minded degradation property, suitable for the waste water containing benzothiazole, give up
The biological treating process of gas, the water of pollution and soil.
A kind of method that CN104001712A discloses alkali thatch and microbial association remedying oil-polluted soils, step is such as
Under：Oil degradation microbial inoculum and soil are pressed 1 with weight ratio:5 ratios are uniformly mixed, and are sprinkled into alkali thatch seed, and plant germination is treated in watering,
It requires to manage according to plant normal growth；The oil degradation microbial inoculum presses 1 for Pseudomonas stutzeri and corynebacterium glutamicum:
1 mixing；Described for the soil of planting alkali thatch is contaminated soil that total petroleum hydrocarbon mass fraction is 10000+5000mg/kg.This
Method is that microbial inoculum and mixing are added in the salt affected soil of oil pollution, then sows alkali thatch seed, periodically waters so that plant
With microorganism Synergistic degradation soil petrochina polluter.Alkali thatch is in addition to playing an important role of to promote petroleum hydrocarbon degradation, also with green
The effect of change adds in microbial bacterial agent and degradation rate and degradation speed is all improved, and what is obtained after microbial degradation has
As vegetable fertilizer, plant and microorganism mutualism, degradation time continues longer machine matter so that petroleum hydrocarbon class pollutant matter has
There is preferable removal effect.
CN104276738A provides a kind of fast and harmless biodegradation method for wax greasy dirt, passes through preparation
Seed liquor, the scattered wax greasy dirt of heating prepare degradation system and biodegradation to realize, are produced specifically for oil recovery oil refining process
Raw wax greasy dirt, the substance in the wax greasy dirt that can effectively degrade, method of the invention is with pseudomonas aeruginosa NY3
It is bacterium source with Pseudomonas stutzeri N2, the hydrocarbon in the greasy dirt that can effectively degrade converts part petroleum hydrocarbon
For glycoprotein substance, the requirement of greasy dirt harmless treatment is finally reached.Using biological treatment, secondary dirt is not generated
Dye, does not have an impact environment；Treatment process is simple, and operating cost is low；Processing time is short, it is efficient.Greasy dirt converted product
There is facilitation to the degradation of greasy dirt, this is a unexpected effect during the present invention tests.
CN105254144A discloses a kind of innoxious oil soil treatment technique, including greasy filth fluidization, biological treatment, sludge
Dehydration and biology four steps of induction, the treatment process steps are that the Candida lipolytica of guarantor is added into biological reactor
Y-57 and pseudomonas putida P-101, after aeration and biological reacts, greasy filth oil-containing is less than 3% in terms of over dry mud, and water oil-containing is small
In 130mg/L；On water surface top, oil slick is recycled by oil-collecting device, and bottom fine deposit standard discharge, greasy filth enters sludge
Dehydration.Microorganism treatment and biological revulsion and Mechanical Method use in conjunction can reach sludge oil content by the present invention
0.5% standard recovers sludge environment, achievees the purpose that recycling, innoxious.
CN106565066A discloses a kind of oily sludge harmless biological processing method, and method is as follows：1)
Crude oil liquefacient is added, solves solidifying crude oil；2) auxiliary material, anaerobism compost are added；3) microbial bacterial agent, aerobic fermentation are added；4)
Plant assistant degradation；Wherein microbial bacterial agent includes the green pus bar of pseudomonas, the micrococcus luteus of bacterium Micrococcus, gemma
The bacillus subtilis of Bacillus, the Acinetobacter junii of acinetobacter, the acetobacter aceti of Acetobacter, weight percent
Ratio is 1:1.5:1:1.6:2.The object of the present invention is to provide a kind of economy, environmental protection, efficient process for treating oil-containing sludge.
CN107299066A discloses preparation method and the degradation treatment side of a kind of microbial degradation liquid of oily sludge
Method, the preparation method include the following steps：S1：The first culture medium of preparing microorganism flora；S2：By complex microbial community
It is cultivated in first culture medium of step S1, obtains the first microflora degradation liquid, the complex microbial community is
Radioresistance acinetobacter calcoaceticus, bacillus subtilis, Acinetobacter lwoffii, Acinetobacter junii, Pseudomonas amygdali and branch's pole
Bacterium is according to weight ratio 1:1-2:0.3-0.7:1-1.4:1:2-3 is mixed；S3：The second culture medium of preparing microorganism flora；S4：
The first microflora degradation liquid is inoculated in second culture medium and is cultivated, obtains the second microflora degradation liquid；S5：It uses
The second microflora degradation liquid is come oily sludge of degrading, completion microbial degradation processing.The method passes through unique microorganism
The composite portfolio of multiple technical characteristics such as flora selection, cultural method and degradation technique operation and collaboration, it is good so as to achieve
Degradation effect.
Bioremediation is divided to two classes, first, adding nutriment into oily sludge, is aerated, and promotes the micro- life of sludge original inhabitants
Object growing multiplication, so as to fulfill the degradation to pollutant.Second is that the microorganism of efficient degradation petroleum hydrocarbon is added into oily sludge
Microbial inoculum.The advantages of biologic treating technique, is：When secondary pollution will not be formed or pollutant is caused to shift；Second is that technique is simple
Single, processing cost is low；Third, high treating effect, by biochemical treatment, pollutant residual quantity can be greatly lowered.Study table
Bright, adding petroleum hydrocarbon degradation bacterium can make degradation rate be increased to more than 50%.Using being metabolized surfactant microorganism to containing greasy dirt
Mud processing, improves petroleum hydrocarbon solubility, can improve the degradation efficiency of petroleum hydrocarbon, can remove PAHs classes using biological treatment has
Evil substance, biological treatment can degrade aliphatic hydrocarbon and aromatic hydrocarbon in oily sludge simultaneously, but bioremediation exists by environment
Factor influences the limitations such as big, long processing period, therefore there are no inputs at home for the treatment technology of biological treatment oily sludge
Scale application.Meanwhile current microbial degradation microbial inoculum, based on complex micro organism fungicide, a variety of bacterial strains need individually culture
It is mixed in a certain ratio after activation, then degrades after carrying out second of culture activation as degradation composite bacteria agent to pollutant,
Culture process is cumbersome.Also there is no more excellent oily sludge petrochina is realized using single strain in the prior art
The technical solution of the degradation of hydro carbons.
The content of the invention
It is an object of the present invention to provide one plant of hydrocarbon degradation bacteria strain, and by it in oily sludge degrades repair process
Using.
One plant of Pseudomonas stutzeri of the present invention and its application in oily sludge of degrading, technical solution are：
A kind of hydrocarbon degradation bacteria strain, which is characterized in that the bacterial strain is Pseudomonas stutzeri category (Pseudomonas
Stutzeri), it is preserved in China General Microbiological culture presevation administrative center (abbreviation CGMCC), address on December 1st, 2017
For Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, preserving number CGMCC
NO.14975。
A kind of efficient degrading bacterial strain that separated as described above from the oily sludge of Liaohe Oil Field is come in oily sludge of degrading
Petroleum hydrocarbon class pollutant method, it is characterised in that specifically implement according to following steps：
Liaohe Oil Field oily sludge 10g is taken to be put into the conical flask equipped with 100mL LB fluid nutrient mediums, 30 DEG C,
It is shaken on the constant-temperature table of 120rpm, enrichment culture is for 24 hours.
The component of the LB culture mediums is：Tryptone 10g, yeast extract 5g, sodium chloride 10g, distilled water 1000mL.
In screening and culturing medium, the initial n-octadecane of screening and culturing medium adds upper strata bacterium solution after inoculation is stood afterwards for 24 hours
Enter amount for 1g/L, the n-octadecane content in screening and culturing medium is continuously improved thereafter until without colony growth in 5d.
The component of the minimal medium is：NaCl 30g, NH4NO33g, KH2PO41g, K2HPO41g,
CaCl20.02g, MgSO40.5g, agar powder 20g；Deionized water 1L；Trace element solution 10ml.
Trace element solution：CuSO40.05g, MnSO40.05g, FeSO4·7H2O 0.05g, deionized water 50ml.
Screening and culturing medium (alkane culture medium)：N-octadecane is added in minimal medium as sole carbon source.
After bacterium colony is grown, colonial morphology is observed, with aseptic inoculation ring from the sieve of the previous concentration of above-mentioned no colony growth
It selects and poor morphology is selected in culture medium away from big bacterium colony, be inoculated in using plate streaking partition method on LB solid mediums, be placed in perseverance
Culture is inverted in warm incubator for 24 hours for 30 DEG C, the picking single bacterium colony after bacterium colony is grown, and the training of LB solids is seeded in the same way
Support base.Above-mentioned line separation process is repeated, until forming the single purifying bacterium colony of form, is inoculated in Storaged media inclined-plane
On saved backup in 4 DEG C of refrigerator.
The colonial morphology of above-mentioned bacterial strains is：Bacterium colony is in faint yellow, and translucent, surface slightly has gloss, and colony edge is unintelligible,
Such as Fig. 1.Thalline is in rod-short, and ne ar is shown in scanning electron microscope Fig. 2.
The 16S ribosomal rna gene sequence signatures of above-mentioned bacterial strains are carried out sequence and database using analytic approach
Analysis is compared, it is found that the bacterial strain belongs to Pseudomonas stutzeri category (Pseudomonas stutzeri), DNA sequence dna table such as sequence
Shown in table.
Bacterium solution is made for 24 hours using preceding activate it in LB fluid nutrient mediums, bacterium solution is inoculated in 10% inoculum concentration
In oily sludge.
Preferably, the optimum condition of the degradation is：10% inoculum concentration, 35 DEG C of temperature, pH degrade under conditions of being 7
Best results.
The present invention has the following advantages compared with prior art：
1st, Pseudomonas stutzeri category (Pseudomonas stutzeri) JN4 of the invention comes from oily sludge, therefore should
Bacterial strain is quickly become dominant microflora after entering oily sludge, occupies certain ecological niche, effectively facilitates the degradation of petroleum hydrocarbon, optimization
Edaphon system improves the physical behavior of soil, enhances the bioactivity of soil.
2nd, Pseudomonas stutzeri category (Pseudomonas stutzeri) JN4 of the invention is using n-octadecane as sole carbon
The dominant strain that source screens, bacterial strain can change the property of oily sludge petrochina hydrocarbons during growth and breeding,
The component of oily sludge petrochina hydro carbons is made to change.
3rd, Pseudomonas stutzeri category (Pseudomonas stutzeri) JN4 of the invention with single bacterial strain to n-C10~n-
C30With apparent degradation capability, by GC-MS analysis shows, the bacterial strain to aromatic hydrocarbon naphthalane, indenes alkane, d-homoandrostane degradation energy
Power is stronger, reaches 100%.
4th, Pseudomonas stutzeri category (Pseudomonas stutzeri) JN4 of the invention be with single strain realize compared with
For the degradation of excellent oily sludge petrochina hydro carbons, reduce each bacterial strain in mix bacterium agent preparation process and activate respectively and in proportion
The complex steps of mixing, the optimum reaction condition of easily controllable oily sludge petroleum hydrocarbon degradation, and single strains for degrading is kept away
It is complicated to exempt from the reaction product after degradation, is easy to joint other methods and further degrades to the hydrocarbon in oily sludge.
Description of the drawings
Fig. 1 is group's form of bacterial strain JN4.
Fig. 2 is the scanning electron microscope (SEM) photograph of bacterial strain JN4.
Fig. 3 Pseudomonas stutzeri JN4 are to the degradation curve of oily sludge petrochina hydro carbons.
Fig. 4 Pseudomonas stutzeri JN4 are under different pH to the degradation curve of oily sludge petrochina hydro carbons.
Fig. 5 Pseudomonas stutzeri JN4 are bent to the degradation of oily sludge petrochina hydro carbons at different temperatures
Line.
Specific embodiment
For a better understanding of the present invention, present invention is further elaborated with reference to embodiment, but
Present disclosure is not limited solely to the following examples.
A kind of degradation bacteria strains of oily sludge petrochina hydro carbons, bacterial strain are Pseudomonas stutzeri category.
(1) material prepares
Fetch oily sludge from Liaohe Oil Field somewhere, 4 DEG C of preservations, transport back be inoculated into LB culture mediums behind laboratory into
Row activation.
2. culture medium：
The component of minimal medium is：N-octadecane 1g, NaCl 30g, NH4NO33g, KH2PO41g, K2HPO41g,
CaCl20.02g, MgSO40.5g, agar powder 20g；Deionized water 1L；Trace element solution 10ml.
Trace element solution：CuSO40.05g, MnSO40.05g, FeSO4·7H2O 0.05g, deionized water 50ml.
LB solid mediums：Tryptone 10g, yeast extract 5g, NaCl 10g, 20 distilled water 1000mL of agar powder,
It is 7 to adjust pH
3. experimental instruments
Constant temperature oscillator
Electro-heating standing-temperature cultivator
High-pressure sterilizing pot,
UV-1750 ultraviolet-uisible spectrophotometers --- -- Prism Optical Technology Co,
Superclean bench,
480 infrared spectrometric oil detectors of OIL --- -- Beijing Kechuang Co., Ltd of China,
PCR instrument.
(2) separation screening of bacterial strain and domestication
Oily sludge 10g is taken to be put into the conical flask equipped with 100ml LB fluid nutrient mediums, in 30 DEG C, the constant temperature of 120rpm
It wants to be shaken on bed, enrichment culture is for 24 hours；
1ml culture solutions is taken to be applied to spreading rod in the solid inorganic salt culture medium containing n-octadecane afterwards for 24 hours, are put into perseverance
Warm incubator culture 7d；
After bacterium colony is grown, colonial morphology is observed, shape is selected from above-mentioned solid inorganic salt culture medium with aseptic inoculation ring
The big bacterium colony of state gap, is inoculated in using plate streaking partition method on LB solid mediums, is placed in constant incubator to fall for 30 DEG C
Put culture for 24 hours, the picking single bacterium colony after bacterium colony is grown, and LB solid mediums are seeded in the same way.Repeat above-mentioned stroke
Line separation process, until forming the single purifying bacterium colony of form.Single bacterium colony is expanded in LB liquid medium numerous, and be stored in 4
DEG C, it is spare.It is activated using the preceding bacterial strain by preservation in LB fluid nutrient mediums.
It is specific embodiment part below.
Embodiment 1：The degradation repairing test of Liaohe Oil Field oily sludge
Liaohe Oil Field oily sludge is taken, grinding is air-dried, crosses 1mm sieves, it is spare.Picking single bacterium falls within the LB Liquid Cultures of sterilizing
It in base, is sealed with breathable sealing film, conical flask is placed in shaking table in 120rpm, 30 DEG C of activation for 24 hours.Weigh above-mentioned air-dried grinding
Oily sludge 40g after sieving, the Pseudomonas stutzeri JN4 that OD600=1 is added according to 10% inoculum concentration live
The bacterium solution of change, and it is 2 to adjust soil ratio with distilled water:5, it is sealed with breathable sealing film.All conical flasks are positioned over 30 DEG C
Quiescent culture in constant incubator is sampled every 5d, and measuring total petroleum hydrocarbon class in oily sludge using infrared spectrometric oil detector contains
Amount, degradation curve is as shown in Figure 3.The bacterium has the petroleum hydrocarbon in oily sludge preferable degradation to imitate as seen from the figure
Fruit was 68.37% to the degradation rate of total petroleum hydrocarbon in oily sludge (TPH) in 30 days.
The GC-MS analysis results of oily sludge before and after Pseudomonas stutzeri JN4 degrade 30 days can be seen that
(table 1), the main component in original oily sludge is naphthalane, n-tetradecane, octadecylene, tetramethyl-pentadecane, tetramethyl ten
Six alkane, indenes alkane, d-homoandrostane, N- (2- trifluoromethylbenzenes)-Niacinamide oxime (C13H10F3N3O), cerotene, hopance.
Contain pregnane (C in oily sludge after Pseudomonas stutzeri JN4 microbial degradations21H36) and Beta-methyl violet
Ketone (C14H22O), illustrate that the variation of molecular structure has occurred in the hydrocarbon in oily sludge under microbial action, greatly
The polluter of molecular structure is under the action of microorganism by oxygenolysis into small molecule structure.Containing after microbial degradation simultaneously
Trimethyl hexadecane and hexadecane in greasy dirt illustrate Pseudomonas stutzeri JN4 microorganisms and tetramethyl hexadecane
Methyl first react, further degrade after demethyl.Pseudomonas stutzeri JN4 are with single bacterial strain to n-
C10~n-C30With apparent degradation capability, and the bacterial strain is to the naphthalane (C in oily sludge10H18), the tetradecane, octadecylene, four
Methyl pentadecane, indenes alkane (C19H36), d-homoandrostane (C21H36O2)、C13H10F3N3The degradation capability of O and cerotene is stronger, drop
Solution rate is up to 100%.After Pseudomonas stutzeri JN4 degrade 30 days, still there is part hydrocarbon in oily sludge
Residual can extend to increase the microbial degradation time and change the modes such as microbial Degradation Conditions and further degrade.
1 Pseudomonas stutzeri JN4 of table degrade 30 days before and after oily sludge GC-MS results
Embodiment 2：Pseudomonas stutzeri JN4 are at various ph values to the drop of oily sludge petrochina hydro carbons
Solve effect
Liaohe Oil Field oily sludge is taken, grinding is air-dried, crosses 1mm sieves.Above-mentioned oily sludge 40g is weighed in conical flask, is used
Dilute H2SO4The pH that soil is adjusted with NaOH is respectively 1,3,5,7,9,11, adds in OD600=1's according to 10% inoculum concentration
The bacterium solution of Pseudomonas stutzeri JN4 activation, and it is 2 to adjust soil ratio with distilled water:5, it is sealed with breathable sealing film
Mouthful.All conical flasks are positioned over quiescent culture in 30 DEG C of constant incubator and were sampled after 25 days, are surveyed using infrared spectroscopy
Oily instrument measures total petroleum hydrocarbon class content in oily sludge.The strain Pseudomonas stutzeri JN4 couple under condition of different pH
The degradation effect of petroleum hydrocarbon in oily sludge is as shown in Figure 4.As shown in Figure 4, bacterial strain of the present invention is in pH 5-11 scopes
The interior petroleum hydrocarbon in oily sludge has preferable removal effect, and degradation effect is optimal under conditions of pH 7.
Embodiment 3：Pseudomonas stutzeri JN4 are at different temperatures to the drop of oily sludge petrochina hydro carbons
Solve effect
Liaohe Oil Field oily sludge is taken, grinding is air-dried, crosses 1mm sieves.Above-mentioned oily sludge 40g is weighed in conical flask, is pressed
According to the bacterium solution that the Pseudomonas stutzeri JN4 of 10% inoculum concentration addition OD600=1 are activated, and adjusted with distilled water
Soil ratio is 2:5, it is sealed with breathable sealing film.By conical flask be respectively placed in 10 DEG C, 15 DEG C, 20 DEG C, 25 DEG C, 30 DEG C, 35 DEG C,
40 DEG C, 45 DEG C, quiescent culture in 50 DEG C of constant incubator, and sampled after 25 days, contained using infrared spectrometric oil detector measure
Total petroleum hydrocarbon class content in greasy dirt.Strain Pseudomonas stutzeri JN4 are in oily sludge at different temperatures
The degradation effect of total petroleum hydrocarbon class is as shown in Figure 5.As shown in Figure 5, bacterial strain of the present invention in the range of 25-40 DEG C to containing greasy dirt
Petroleum hydrocarbon in mud has certain degradation effect, and degradation rate is then reduced as the rise of temperature first raises, 35 DEG C of items
It degrades under part best results.
Embodiment 4：Pseudomonas stutzeri belongs to Pseudomonas stutzeri JN4 and repairs actual oily sludge
On the basis of above example, Pseudomonas stutzeri is belonged into Pseudomonas stutzeri JN4 for repairing
Actual oily sludge, experiment are as follows：
(1) prepared by the bacteria suspension of strain Pseudomonas stutzeri JN4：The single bacterium of picking bacterial strain JN4 falls within LB liquid
It in body culture medium, is positioned in shaking table, in 40 DEG C, is cultivated for 24 hours under the conditions of 120rpm.Fermented and cultured is carried out using fermentation tank, it will
The seed liquor of above-mentioned culture is inoculated in by 5% inoculum concentration in LB culture mediums, is aerated using air compressor, speed of agitator
For 200r/min, timing sampling detection OD600 values are grown to spare after exponential phase.
(2) for examination oily sludge：Liaohe Oil Field somewhere oily sludge is picked up from, through natural air drying, crosses 8 mesh sieves (3mm sieves).It will
Oily sludge sample is fitted into plexiglass box, and the Pseudomonas of OD600=1 is added according to 10% inoculum concentration
The bacterium solution of stutzeri JN4 activation, and liquid soil is adjusted than being 2 with minimal medium:5, it is sealed with breathable sealing film.
(3) oily sludge for being with the addition of bacterium solution and minimal medium is positioned over quiescent culture in constant incubator, is
Reach optimal degradation effect, the pH value for adjusting soil is 7, and maintains temperature at 35 DEG C.
Oily sludge is detected after the biodegradable processing of 30 days, is found to total petroleum hydrocarbon in oily sludge
(TPH) degradation rate is 71.05%, has preferable degradation effect to the petroleum hydrocarbon in oily sludge.
Sequence table
<110>Peking University
<120>A kind of degradation bacteria strains JN4 of oily sludge petrochina hydro carbons and its application
<130> 2018
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1410
<212> DNA
<213>Pseudomonas stutzeri (Pseudomonas stutzeri)
<400> 1
tgcagtcgag cggatgaagg gagcttgctc cctgatttag cggcggacgg gtgagtaatg 60
cctaggaatc tgcctggtag tgggggacaa cgtttcgaaa ggaacgctaa taccgcatac 120
gtcctacggg agaaagcagg ggaccttcgg gccttgcgct atcagatgag cctaggtcgg 180
attagctagt tggtgaggta aaggctcacc aaggcgacga tccgtaactg gtctgagagg 240
atgatcagtc acactggaac tgagacacgg tccagactcc tacgggaggc agcagtgggg 300
aatattggac aatgggcgaa agcctgatcc agccatgccg cgtgtgtgaa gaaggtcttc 360
ggattgtaaa gcactttaag ttgggaggaa gggcagtaag ctaatacctt gctgttttga 420
cgttaccgac agaataagca ccggctaact tcgtgccagc agccgcggta atacgaaggg 480
tgcaagcgtt aatcggaatt actgggcgta aagcgcgcgt aggtggttcg ttaagttgga 540
tgtgaaagcc ccgggctcaa cctgggaact gcatccaaaa ctggcgagct agagtatggc 600
agagggtggt ggaatttcct gtgtagcggt gaaatgcgta gatataggaa ggaacaccag 660
tggcgaaggc gaccacctgg gctaatactg acactgaggt gcgaaagcgt ggggagcaaa 720
caggattaga taccctggta gtccacgccg taaacgatgt cgactagccg ttgggatcct 780
tgagatctta gtggcgcagc taacgcatta agtcgaccgc ctggggagta cggccgcaag 840
gttaaaactc aaatgaattg acgggggccc gcacaagcgg tggagcatgt ggtttaattc 900
gaagcaacgc gaagaacctt accaggcctt gacatgcaga gaactttcca gagatggatt 960
ggtgccttcg ggaactctga cacaggtgct gcatggctgt cgtcagctcg tgtcgtgaga 1020
tgttgggtta agtcccgtaa cgagcgcaac ccttgtcctt agttaccagc acgttaaggt 1080
gggcactcta aggagactgc cggtgacaaa ccggaggaag gtggggatga cgtcaagtca 1140
tcatggccct tacggcctgg gctacacacg tgctacaatg gtcggtacaa agggttgcca 1200
agccgcgagg tggagctaat cccataaaac cgatcgtagt ccggatcgca gtctgcaact 1260
cgactgcgtg aagtcggaat cgctagtaat cgtgaatcag aatgtcacgg tgaatacgtt 1320
cccgggcctt gtacacaccg cccgtcacac catgggagtg ggttgctcca gaagtagcta 1380
gtctaacctt cgggaggacg gtaccacgga 1410
Claims (8)
1. the bacterial strain JN4 that a kind of oily sludge degradation is repaired, which is characterized in that the bacterial strain is Pseudomonas stutzeri category
Pseudomonas stutzeri, the entitled China General Microbiological culture presevation administrative center of depositary institution, preserving number are
CGMCC NO.14975。
2. the bacterial strain JN4 that a kind of oily sludge degradation according to claim 1 is repaired, it is characterised in that：The degradation bacteria
The purposes that the degradation of oily sludge is repaired is carried out in a manner that petroleum hydrocarbon is degraded.
3. the bacterial strain JN4 that a kind of oily sludge degradation according to claim 1 is repaired, it is characterised in that：The degradation bacteria
Degradation condition to petroleum hydrocarbon is 25-40 DEG C, pH value 5-11.
4. the bacterial strain JN4 that a kind of oily sludge degradation according to claim 1 is repaired, it is characterised in that：The degradation bacteria
With single bacterial strain to n-C10~n-C30With apparent degradation capability, by GC-MS analysis shows, the bacterial strain is to aromatic hydrocarbon naphthalene
Alkane, indenes alkane, the degradation capability of d-homoandrostane are stronger, reach 100%.
5. the application for the bacterial strain JN4 that a kind of oily sludge degradation according to claim 2 is repaired, it is characterized in that, including such as
Lower step：
(1) picking Pseudomonas stutzeri JN4 single bacteriums are fallen in the conical flask of LB fluid nutrient mediums of sterilizing, with saturating
Sealing gland membrana oralis seal, by conical flask be placed in shaking table in 120rpm, 30 DEG C, activation seed culture fluid is made for 24 hours.
(2) seed liquor of above-mentioned culture is inoculated in culture medium fermentation tank containing LB by 5% inoculum concentration to cultivate, using air
Compressor is aerated, and speed of agitator 200r/min is cultivated to exponential phase.
(2) by Liaohe Oil Field oily sludge, air-dried grinding after crossing 1mm sieves, adds in OD600=1's according to 10% inoculum concentration
The bacterium solution of Pseudomonas stutzeri JN4 activation, and liquid soil is adjusted than being 2 with minimal medium:5, use breathable sealing
Film seals.
(3) oily sludge for being with the addition of bacterium solution and minimal medium is positioned over quiescent culture in constant incubator, to reach
Optimal degradation effect, the pH value for adjusting soil is 7, and maintains temperature at 35 DEG C.
6. the application for the bacterial strain JN4 that a kind of oily sludge degradation according to claim 5 is repaired, it is characterized in that：Described
The component of LB culture mediums is：Tryptone 10g, yeast extract 5g, NaCl 10g, distilled water 1000mL, it is 7 to adjust pH.
7. the application for the bacterial strain JN4 that a kind of oily sludge degradation according to claim 5 is repaired, it is characterized in that：Described
The component of minimal medium is：NaCl 30g, NH4NO33g, KH2PO41g, K2HPO41g, CaCl20.02g, MgSO4
0.5g, agar powder 20g；Deionized water 1L；Trace element solution 10ml.
8. the component of minimal medium according to claim 6, it is characterized in that：The component of trace element solution is：
CuSO40.05g, MnSO40.05g, FeSO4·7H2O 0.05g, deionized water 50ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810115860.8A CN108048374B (en) | 2018-02-06 | 2018-02-06 | Degradation strain JN4 for petroleum hydrocarbons in oily sludge and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810115860.8A CN108048374B (en) | 2018-02-06 | 2018-02-06 | Degradation strain JN4 for petroleum hydrocarbons in oily sludge and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108048374A true CN108048374A (en) | 2018-05-18 |
| CN108048374B CN108048374B (en) | 2020-08-04 |
Family
ID=62125649
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810115860.8A Active CN108048374B (en) | 2018-02-06 | 2018-02-06 | Degradation strain JN4 for petroleum hydrocarbons in oily sludge and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108048374B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109482638A (en) * | 2018-12-14 | 2019-03-19 | 中国石油集团川庆钻探工程有限公司 | Pseudomonas oil-reducing bacterium L-1, mixed microbial inoculum thereof and in-situ remediation method for petroleum-polluted soil |
| CN109576186A (en) * | 2018-12-27 | 2019-04-05 | 天津天丰泽田生物科技有限公司 | A kind of petroleum hydrocarbon degradation composite bacteria agent and preparation method thereof |
| CN111410392A (en) * | 2020-04-30 | 2020-07-14 | 袁登辉 | Biological treatment method for removing oil in oily sludge |
| CN115415299A (en) * | 2022-08-29 | 2022-12-02 | 东华大学 | Method for repairing pyrene contaminated soil through low-temperature heat treatment-pyrene degradation microbial inoculum combination |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102250798A (en) * | 2011-06-21 | 2011-11-23 | 中国农业科学院农业环境与可持续发展研究所 | Pseudomonas and microbial agent as well as applications thereof |
| CN103981119A (en) * | 2014-02-28 | 2014-08-13 | 天津科技大学 | Petroleum efficient degrading bacterium and bacterium group both in oily sludge and application of two |
| CN104745506A (en) * | 2015-03-11 | 2015-07-01 | 河南省水文地质工程地质勘察院有限公司 | Petroleum hydrocarbon degrading bacteria and application thereof |
| CN105176888A (en) * | 2015-10-22 | 2015-12-23 | 中国石油大学(华东) | Pseudomonad strain and application thereof in degrading petroleum pollutants |
-
2018
- 2018-02-06 CN CN201810115860.8A patent/CN108048374B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102250798A (en) * | 2011-06-21 | 2011-11-23 | 中国农业科学院农业环境与可持续发展研究所 | Pseudomonas and microbial agent as well as applications thereof |
| CN103981119A (en) * | 2014-02-28 | 2014-08-13 | 天津科技大学 | Petroleum efficient degrading bacterium and bacterium group both in oily sludge and application of two |
| CN104745506A (en) * | 2015-03-11 | 2015-07-01 | 河南省水文地质工程地质勘察院有限公司 | Petroleum hydrocarbon degrading bacteria and application thereof |
| CN105176888A (en) * | 2015-10-22 | 2015-12-23 | 中国石油大学(华东) | Pseudomonad strain and application thereof in degrading petroleum pollutants |
Non-Patent Citations (2)
| Title |
|---|
| 任丽君，刘宪斌，田胜艳: "含油污泥中石油降解菌的分离及其降解特性", 《环境工程学报》 * |
| 杨丞磊，罗宇维，陈平，朱战兵，项先忠，杨知勋，鲁军: "石油烃降解菌的初筛及其降解性能研究", 《化学工程师》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109482638A (en) * | 2018-12-14 | 2019-03-19 | 中国石油集团川庆钻探工程有限公司 | Pseudomonas oil-reducing bacterium L-1, mixed microbial inoculum thereof and in-situ remediation method for petroleum-polluted soil |
| CN109576186A (en) * | 2018-12-27 | 2019-04-05 | 天津天丰泽田生物科技有限公司 | A kind of petroleum hydrocarbon degradation composite bacteria agent and preparation method thereof |
| CN111410392A (en) * | 2020-04-30 | 2020-07-14 | 袁登辉 | Biological treatment method for removing oil in oily sludge |
| CN111410392B (en) * | 2020-04-30 | 2023-05-02 | 袁登辉 | Biological treatment method for removing oil in oily sludge |
| CN115415299A (en) * | 2022-08-29 | 2022-12-02 | 东华大学 | Method for repairing pyrene contaminated soil through low-temperature heat treatment-pyrene degradation microbial inoculum combination |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108048374B (en) | 2020-08-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109207413B (en) | A kind of highly effective petroleum degradation composite bacteria agent and the preparation method and application thereof | |
| CN101580808B (en) | Rhodococcus ruber and application thereof in degradation of hydrocarbon compounds | |
| de-Bashan et al. | The potential contribution of plant growth-promoting bacteria to reduce environmental degradation–A comprehensive evaluation | |
| CN110257310B (en) | A kind of vinyon efficient degrading bacteria and its separating screening method and application | |
| CN108102979A (en) | A kind of degradation bacteria strains JN5 of oily sludge petrochina hydro carbons and its application | |
| CN103436464B (en) | Low temperature-resistant petroleum-degrading bacillus sp. strain, culture method and application thereof | |
| CN108048374A (en) | A kind of degradation bacteria strains JN4 of oily sludge petrochina hydro carbons and its application | |
| CN103981119B (en) | The application of oily sludge petrochina efficient degrading bacteria and bacterium group | |
| CN102533619B (en) | Preparation method of hydrochloric ether efficient aerobic degradation mixed bacteria and application thereof | |
| CN110283772A (en) | A kind of preparation method of functional flora that repairing petroleum hydrocarbon contaminated soil and underground water | |
| CN111748483B (en) | Bacillus for degrading petroleum hydrocarbon and application thereof | |
| CN108034626A (en) | A kind of degradation bacteria strains JN1 of oily sludge petrochina hydro carbons and its application | |
| CN103215204A (en) | Arthrobacter strain for efficiently degrading phenanthrene and application thereof | |
| JPWO2020009097A1 (en) | Decontamination methods and materials used for environments contaminated by petroleum-related substances | |
| CN111733098B (en) | Application of bacillus in low-temperature degradation of petroleum hydrocarbon | |
| CN105647838B (en) | Skin spy's acinetobacter calcoaceticus and application thereof | |
| CN108102978A (en) | A kind of degradation bacteria strains JN8 of oily sludge petrochina hydro carbons and its application | |
| CN101935631B (en) | Ralstoniasp. and application thereof in bioremediation of petroleum-contaminated saline-alkali soil | |
| CN106497818B (en) | Pseudomonas aeruginosa and microbial inoculum and their applications in degraded oil and/or oil product | |
| CN108277175A (en) | 2,4 dinitrotoluene (DNT) sulfonate efficient degrading bacterial strain Microbacterium sp.X3 and its application | |
| CN114854626A (en) | Pseudomonas strain for degrading polycyclic aromatic hydrocarbon pollutants and application thereof | |
| CN108034625A (en) | A kind of degradation bacteria strains JN7 of oily sludge petrochina hydro carbons and its application | |
| CN109455891A (en) | A kind of compound bio preparation for repairing for oily sludge | |
| CN107129940A (en) | Composite flora, microbial inoculum and the culture of remedying oil-polluted soils, process for fixation and application | |
| CN114058528A (en) | Oily pollutant degradation strain XT4, microbial agent and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |