CN108298780A - A kind of biological cleaner and application method of processing oily sludge - Google Patents
A kind of biological cleaner and application method of processing oily sludge Download PDFInfo
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- CN108298780A CN108298780A CN201810053678.4A CN201810053678A CN108298780A CN 108298780 A CN108298780 A CN 108298780A CN 201810053678 A CN201810053678 A CN 201810053678A CN 108298780 A CN108298780 A CN 108298780A
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- rhamnolipid
- biological cleaner
- oily sludge
- biological
- cleaner
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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
- C02F11/002—Sludge treatment using liquids immiscible with water
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- Hydrology & Water Resources (AREA)
- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to field of solid waste treatment, more particularly to a kind of biological cleaner and application method of processing oily sludge.Biological cleaner active constituent is single rhamnolipid, wherein a concentration of 0.01wt%~0.2wt% of single rhamnolipid in biological cleaner.The greasy filth cleaning agent of the present invention is environmentally protective, preparation is easy, cleaning performance is good, has a wide range of application, and low energy consumption for the cleaning method based on the cleaning agent, can be applied to the greasy filth of generation during processing oil exploitation, the defeated, refinery of collection etc..
Description
Technical field
The invention belongs to field of solid waste treatment, more particularly to it is a kind of processing oily sludge biological cleaner and make
Use method.
Background technology
A large amount of oily solid sludge is generated during oil exploitation, transport, refining and oily waste water treatment, it has also become
The important sources of soil petroleum pollution.National annual about 10,000,000 tons of oil-containing mud sand of generation, oil-containing mud sand has been put into《Country's danger
Dangerous waste register》, it is the primary pollutant in oil field.The oil content of greasy filth is between 10%-70%, although greasy filth is a kind of pollution
Object, but clean and reuse crude oil therein is carried out to greasy filth so that greasy filth becomes a kind of resource again.With other oil-sludge treatment methods
It compares, Biochemical method greasy filth has the characteristics that not formed secondary pollution, economic and practical, and treatment effect and environmental benefit have more
Advantage.However greasy filth of the oil content less than 10% could be suitble to microbial degradation to handle.Develop environment-friendly cleaning agent to greasy filth into
Row cleaning, on the one hand can recycle crude resources, and on the other hand can make that greasy filth oil content is reduced to microbial degradation processing again can
The scope of application.
Currently, mainly being cleaned to greasy filth using chemical surfactant, although chemical surfactant can be preferable
Oily sludge is cleaned, but needs higher concentration and treatment temperature, and the biological degradability of chemical surfactant is poor, has one
Determine toxicity, secondary pollution is easily caused to environment.Therefore, obtain it is a kind of environmentally protective and can cleaning mud oil at normal temperatures cleaning
Agent will solve above-mentioned bottleneck problem.
Invention content
In order to overcome the above-mentioned deficiencies of the prior art, the present invention provides a kind of environmentally protective and can clean at normal temperatures
The biological cleaner and application method of a kind of processing oily sludge of greasy filth.
To achieve the above object, the invention adopts a technical scheme as：
A kind of biological cleaner of processing oily sludge, biological cleaner active constituent are single rhamnolipid, wherein biology
A concentration of 0.01wt%~0.2wt% of single rhamnolipid in cleaning agent.
A concentration of 0.01wt%~0.1wt% of single rhamnolipid in the biological cleaner.
The biological cleaner is solution form, is using single rhamnolipid producing strains as the sandlwood containing list obtained by starting strain
Fermentation culture medium, fermented and cultured suspension or the fermentation clear liquid of glycolipid, or the solution containing single rhamnolipid.
The list rhamnolipid is using single rhamnolipid producing strains as fermentation culture medium obtained by the fermented culture of starting strain
Extract, or single rhamnolipid by detaching acquisition in rhamnolipid.
The list rhamnolipid producing strains are the improved bacterial strains of genetic engineering, using pseudomonas aeruginosa as starting strain,
It knocks out and synthesizes relevant key gene rhlC with pair rhamnolipids and it is made not express, or by the key of pseudomonas aeruginosa
Gene rhlAB, which is imported into suitable host strain, carries out heterogenous expression so that is only expressed in two genoid engineering bacterias and single sandlwood
Glycolipid synthesizes relevant key gene rhlAB, finally makes to be complete single rhamnose in the rhamnolipid product of genetic engineering bacterium
Fat, such as Pseudomonas stutzeri Pseudomonas stutzeri Rhl (the related notes of Pseudomonas stutzeri Rhl
It carries referring to Zhao et al.2015 Heterologous production of Pseudomonas aeruginosa
rhamnolipid under anaerobic conditions for microbial enhanced oil recovery)。
By taking Pseudomonas stutzeri Pseudomonas stutzeri Rhl as an example：
Single rhamnolipid is to be inoculated in hair using Pseudomonas stutzeri Pseudomonas stutzeri Rhl as starting strain
In ferment culture medium under the conditions of 30~40 DEG C, 160~220rpm, cultivate 3~6 days, the obtained zymotic fluid containing single rhamnolipid；
The fermentation medium is by mass fraction：Carbon source 3%, NaNO30.3%, KH2PO40.1%, K2HPO4·3H2O 0.2%,
MgSO4·7H2O 0.04%, yeast powder 0.01%, surplus are water；PH 6.5~7.0.
It is described by starting strain seed liquor, be 2-6 by seed liquor and the volume ratio of fermentation medium:100 ratio inoculation.
The carbon source is one or more in glycerine, glucose, sucrose, molasses.
A kind of application method of the biological cleaner of processing oily sludge, the cleaning agent is mixed with pending greasy filth,
The oily sludge of concussion processing at normal temperatures；Wherein, cleaning agent and pending greasy filth solid-liquid mass volume ratio (g/ml) are 1:5-10.
It is described to mix the cleaning agent with pending greasy filth, under the conditions of at 30-45 DEG C with the concussion of 150-200rpm
Handle 12-48h；A concentration of 200-1000mg/L of single rhamnolipid in the biological cleaner.
Beneficial effects of the present invention：
The greasy filth cleaning agent main component of the present invention is single rhamnolipid, water-wet side containing only there are one sandlwood saccharide ring, because
This single rhamnolipid has better oil-wet behavior, shows as having better emulsification of crude oil activity, has the cleaning of preferable greasy filth
Activity also has many advantages, such as that environmental-friendly, preparation is easy, cleaning performance is good.In addition, the cleaning method energy consumption of cleaning agent of the present invention
It is low, greasy filth cleaning can be carried out at normal temperatures.Therefore, biological cleaner and cleaning method of the present invention, recycling, decrement in greasy filth
Changing process field will be with important application value.
Specific implementation mode
In conjunction with following specific embodiments, the present invention will be described in detail, makes those skilled in the art more comprehensively
Understanding the present invention, but do not limit the invention in any way.In following examples, unless otherwise specified, material used and
Reagent can be bought from biochemical reagents company and obtain；In addition to the method for specified otherwise, test method used is laboratory routine
Experimental method.
Embodiment 1：Prepare single rhamnolipid fermentation liquor
It is weighed respectively with electronic balance, 30g glucose, 3g NaNO3, 1g KH2PO4, 2g K2HPO4·3H2O, 0.4g
MgSO4·7H2O, 1g yeast powder；After distilled water mixing is added, 1L is settled to distilled water；The NaOH of mass fraction 30% is added,
Adjust the pH to 7.0 of culture medium；The 30min of sterilizing is placed at 121 DEG C after packing.
Rhamnolipid product is selected to be entirely the Pseudomonas stutzeri Pseudomonas stutzeri of single rhamnolipid
Rhl (record referring to Zhao et al.2015Heterologous by the related of Pseudomonas stutzeri Rhl
production of Pseudomonas aeruginosa rhamnolipid under anaerobic conditions
For microbial enhanced oil recovery) strain as production list rhamnolipid.
After fermentation medium is cooled to room temperature (20-30 DEG C), the single rhamnolipid of generation of inoculation medium volume 3%
Seed liquor (the bacterium number 10 of bacterium Rhl5A/ml), it under the conditions of 37 DEG C, 180rpm, cultivates 5 days, is used for fermenting and producing list sandlwood
Glycolipid obtains single rhamnolipid fermentation liquor.
Wherein, the seed liquor of Rhl is to take 2 oese of inclined-plane of Rhl bacterium, is inoculated into the triangular flask of the culture mediums of LB containing 100ml
In, at 30 DEG C, 180rpm/min, cultivate 6-10 hours, until OD600Value is for 0.7 to get to culture to the logarithmic growth later stage
Seed liquor, bacterium number 105A/ml.
Above-mentioned acquisition list rhamnolipid fermentation liquor is diluted with water, single rhamnolipid concentration is made to reach 100-500mg/
L, with the biological cleaner for dealing with greasy filth.
Embodiment 2：Prepare single rhamnolipid aqueous solution
The zymotic fluid that above-described embodiment culture is terminated is centrifuged off the insoluble matters such as thalline under the conditions of 10,000g, profit
With chloroform/methanol (v/v, 2:1) single rhamnolipid in extraction extraction zymotic fluid.
Qualitative analysis is carried out to the single rhamnolipid product extracted using thin-layered chromatography, takes the sandlwood after a small amount of extraction
Glycolipid product is dissolved in dichloromethane, prepares single rhamnolipid sample of 200mg/L, by sample spot on silica gel plate, chloroform/
Methanol/acetic acid=65:15:It is unfolded in the solvent of 2 (v/v/v), is developed the color with color developing agent.Under phenolsulfuric acid color developing agent,
Product shows yellow, calculates its Rf value.Under phenolsulfuric acid color developing agent, product shows yellow, and on thin layer silica gel plate only
One point, Rf value (Rf values) are 0.82.Exist if obtained product is the mixture of single rhamnolipid and double rhamnolipids
It will appear two different points of Rf values on lamellae.
Under the conditions of the Development of Thin-Layer Chromatography, monosaccharide fat polarity is weak, and the adsorption coefficient between silica gel plate is small and has stronger
Migration performance.It is shown according to documents and materials, the Rf values of single rhamnolipid are about 0.89, the Rf values of double rhamnolipids are about 0.55
(record related to double rhamnolipid thin-layer chromatographic analysis of single rhamnolipid is referring to the thalline of clock China .2008. rhamnolipids is inhaled
Modifying function research attached and its to phage surface).Therefore, illustrate that the rhamnolipid product extracted from zymotic fluid is only
Single rhamnolipid.
Aqueous solution is prepared using the single rhamnolipid extracted, wherein a concentration of 100-500mg/L of single rhamnolipid, is used as
Handle the biological cleaner of greasy filth.
Embodiment 3：Application examples
NW China Sludge in Oilfields is cleaned using the biological cleaner that Examples 1 and 2 obtain respectively
It with electronic balance weighing oily sludge 10g, pours into triangular flask, is separately added into 100ml embodiments 1 or embodiment 2
The biological cleaner of a concentration of 300mg/L obtained, stirs and evenly mixs, under the conditions of 37 DEG C, 180rpm, oscillation cleaning greasy filth 48h.
After cleaning, oil, water, mud separation are carried out, the upper layer oil that recovery and rinsing goes out, the final oil of 1 gained biological cleaner of embodiment returns
Yield is 56.8%；The 2 final oil recovery rate of gained biological cleaner of embodiment is 59.1%.
Embodiment 4：Comparative example
The rhamnolipid product of most rhamnolipid producing strains synthesis is the mixed of single rhamnolipid and double rhamnolipids
Close object.The single rhamnolipid of embodiment comparison and common rhamnolipid (single rhamnolipid and double rhamnolipid mixtures), to oil
The treatment effect of mud.
Selection while the pseudomonas aeruginosa (Pseudomonas for producing single rhamnolipid and double rhamnolipids
aeruginosa)ATCC 10145(Chayabutra et al.,Polyhydroxyalkanoic acids and
rhamnolipids are synthesized sequentially in hexadecane fermentation by
Pseudomonas aeruginosa ATCC 10145.Biotechnology progress,2001,17(3):419-423.)
For starting strain, 2 oese of slant strains of strains A TCC 10145 is taken, the triangular flask of the culture mediums of LB containing 100ml is inoculated into
In, at 37 DEG C, 180rpm/min, cultivate 6-10 hours, until OD600Value is for 0.7 to get to culture to the logarithmic growth later stage
Seed liquor, bacterium number 105A/ml.
Utilize the culture medium in embodiment 1, the seed liquor of the strains A TCC 10145 of inoculation medium volume 3%, 37
DEG C, under the conditions of 180rpm, cultivate 4 days, obtain the zymotic fluid containing rhamnolipid.
By above-mentioned zymotic fluid, under the conditions of 10,000g, be centrifuged off the insoluble matters such as thalline, using chloroform/methanol (v/v,
2:1) the rhamnolipid product in extraction extraction zymotic fluid, i.e., common rhamnolipid.
The common rhamnolipid product of above-mentioned acquisition is diluted with water, rhamnolipid concentration is made to reach 300mg/L, is used
Deal with the biological cleaner A of greasy filth.
The above-mentioned rhamnolipid product being obtained by extraction is subjected to silica gel column chromatography, isolates single rhamnolipid in product.
Silica gel column chromatography separation method can refer to pertinent literature, and (thalline of clock China .2008. rhamnolipids adsorbs and its to phage surface
Modifying function studies).The single rhamnolipid being separated to is verified by thin layer chromatography in embodiment 2.
Single rhamnolipid isolate of above-mentioned acquisition is diluted with water, single rhamnolipid concentration is made to reach 300mg/L,
With the biological cleaner B for dealing with greasy filth.
NW China Sludge in Oilfields is handled under same room temperature cleaning condition using biological cleaner A and B, finally
Oil recovery rate is respectively 20.3% and 55.7%.
Single rhamnolipid of the greasy filth cleaning agent Microbe synthesis of the present invention, environmental-friendly, preparation simplicity, cleaning performance
Good, low energy consumption for the cleaning method based on the cleaning agent, can carry out crude oil washing/COW at normal temperatures.The biological cleaner and cleaning side
Method in recycling, the minimizing process field of greasy filth there are important promotion and application to be worth.
Claims (6)
1. a kind of biological cleaner of processing oily sludge, it is characterised in that：Biological cleaner active constituent is single rhamnolipid,
Wherein, a concentration of 0.01wt%~0.2wt% of single rhamnolipid in biological cleaner.
2. the biological cleaner of processing oily sludge described in accordance with the claim 1, it is characterised in that：In the biological cleaner
Single a concentration of 0.01wt%~0.1wt% of rhamnolipid.
3. according to the biological cleaner of processing oily sludge as claimed in claim 1 or 2, it is characterised in that：The biology cleaning
Agent be using single rhamnolipid producing strains as obtained by starting strain the fermentation culture medium containing single rhamnolipid, fermented and cultured suspension or
Fermentation clear liquid, or the solution containing single rhamnolipid.
4. the biological cleaner of processing oily sludge described in accordance with the claim 3, it is characterised in that：It is described list rhamnolipid be
Using single rhamnolipid producing strains as the extract of fermentation culture medium obtained by the fermented culture of starting strain, or by rhamnolipid
Single rhamnolipid that middle separation obtains.
5. a kind of application method of the biological cleaner of processing oily sludge described in claim 1, it is characterised in that：It will be described
Cleaning agent is mixed with pending greasy filth, the oily sludge of concussion processing at normal temperatures；Wherein, cleaning agent and pending greasy filth solid-liquid matter
It is 1 to measure volume ratio (g/ml):5-10.
6. the application method of the biological cleaner of processing oily sludge as described in claim 5, it is characterised in that：It is described by institute
It states cleaning agent to mix with pending greasy filth, to handle 12-48h under the conditions of the concussion of 150-200rpm at 30-45 DEG C；The life
A concentration of 200-1000mg/L of single rhamnolipid in object cleaning agent.
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| CN201810053678.4A CN108298780B (en) | 2018-01-19 | 2018-01-19 | Biological cleaning agent for treating oily sludge and use method |
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| CN201810053678.4A CN108298780B (en) | 2018-01-19 | 2018-01-19 | Biological cleaning agent for treating oily sludge and use method |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109987806A (en) * | 2019-03-02 | 2019-07-09 | 西南石油大学 | A kind of greasy filth cleaning and recovery method based on food waste grease |
| CN111849846A (en) * | 2020-07-06 | 2020-10-30 | 曲阜师范大学 | Gene modification method for high-yield monorhamnolipid and application thereof |
| CN112205407A (en) * | 2020-09-29 | 2021-01-12 | 曲阜师范大学 | Agricultural bacteriostatic agent from microorganisms and preparation method thereof |
| CN112708510A (en) * | 2020-12-30 | 2021-04-27 | 宁波锋成先进能源材料研究院有限公司 | Cleaning agent and preparation method and application thereof |
| CN115010333A (en) * | 2022-06-27 | 2022-09-06 | 郑州大学 | Oily sludge cleaning fluid and oily sludge treatment method |
| CN116144333A (en) * | 2021-11-23 | 2023-05-23 | 中国石油天然气股份有限公司 | Super-thick oil biochemical viscosity reducer and preparation method thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109987806A (en) * | 2019-03-02 | 2019-07-09 | 西南石油大学 | A kind of greasy filth cleaning and recovery method based on food waste grease |
| CN111849846A (en) * | 2020-07-06 | 2020-10-30 | 曲阜师范大学 | Gene modification method for high-yield monorhamnolipid and application thereof |
| CN112205407A (en) * | 2020-09-29 | 2021-01-12 | 曲阜师范大学 | Agricultural bacteriostatic agent from microorganisms and preparation method thereof |
| CN112708510A (en) * | 2020-12-30 | 2021-04-27 | 宁波锋成先进能源材料研究院有限公司 | Cleaning agent and preparation method and application thereof |
| CN116144333A (en) * | 2021-11-23 | 2023-05-23 | 中国石油天然气股份有限公司 | Super-thick oil biochemical viscosity reducer and preparation method thereof |
| CN116144333B (en) * | 2021-11-23 | 2024-01-30 | 中国石油天然气股份有限公司 | Super-thick oil biochemical viscosity reducer and preparation method thereof |
| CN115010333A (en) * | 2022-06-27 | 2022-09-06 | 郑州大学 | Oily sludge cleaning fluid and oily sludge treatment method |
| CN115010333B (en) * | 2022-06-27 | 2023-09-15 | 郑州大学 | Oily sludge cleaning liquid and treatment method of oily sludge |
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