CN104140962B - A kind of process for fixation of oil degradation flora - Google Patents
A kind of process for fixation of oil degradation flora Download PDFInfo
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- CN104140962B CN104140962B CN201310523187.9A CN201310523187A CN104140962B CN 104140962 B CN104140962 B CN 104140962B CN 201310523187 A CN201310523187 A CN 201310523187A CN 104140962 B CN104140962 B CN 104140962B
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Abstract
The present invention discloses a kind of process for fixation of oil degradation flora, belongs to microorganism fixed network.Specifically, comprising the following steps：（1）Oil degradation flora liquid fermentation；（2）Immobilization of the oil degradation flora on diatomite/activated carbon composite carrier.Oil degradation flora process for fixation proposed by the invention has cost low, beneficial to the high advantage of large-scale application and oil removal efficiency, and the degradation flora synergy of immobilization is strong, degradation capability is big, it is adaptable to the biodegradation of all kinds of oil-containing solid wastes.
Description
Technical field：
The present invention relates to a kind of process for fixation of oil degradation flora, belong to microorganism fixed network.
Background technology：
In recent years, as countries in the world are to oil and its growing demand of product, Exploration of Oil And Gas, exploitation drilling well
During a large amount of oil-containing solid wastes for producing, because it contains the pollutants such as petroleum hydrocarbon, heavy metal and organic matter, be put into state
Family's danger wastes, if just directly being discharged without processing, it will cause to seriously endanger to ecological environment.Therefore, oil-containing solid waste is dirty
Dye has become the great environmental problem that people must face.In oil-containing solid waste treatment technology, biologic treating technique is because of tool
There are safety, economic, efficient, non-secondary pollution, it has also become the important hand of live oil-containing solid waste safe handling and disposal
Section, the hole oil and oil sheet that can not be particularly removed to mechanical device, while when limitation is using chemical agent again, with biology
Treatment technology can show bigger superiority.
Oil is that one kind contains a variety of hydro carbons（Normal alkane, branched paraffin, aromatic hydrocarbons and alicyclic）And other a small amount of organic matters
（Sulfide, nitride and naphthenic）Complex mixture.One typical oil sample contains different hydrocarbon compounds
Up to 200~300 kinds.Different microorganisms has different degradation capabilities to petroleum component, and single oil degradation bacteria is difficult
Oil is thoroughly removed from environment, therefore, the complexity of petroleum components determines that its degraded needs the common of multiple-microorganism
Participate in.The structure of oil degradation flora can make full use of symbiosis and synergy between each degradation bacteria, realize the same of petroleum component
Step degraded, greatly improves the degradation rate and degree of petroleum pollution, effectively shortens the degradation cycle of petroleum pollution.
Immobilized microorganism technology is to be fixed on free microorganism by using method physically or chemically to limit space
In region, retain its intrinsic activity, and the modern biological project technique that can be repeated and continuously used.On oil degradation list
The immobilization technology of bacterium is it has been reported that 200910255830.8 disclose a kind of oil degradation using reed earth's surface coring as carrier
Bacteria immobilization method；201210006059.2 immobilizing oil degradation bacteria method and its application are disclosed, the fixation of both approaches
Change single bacterium, due to limitation of the single bacterium degradation bacteria to petroleum hydrocarbon degradation species, it is impossible to realize the synchronous degradation of petroleum component, cause
Degradation cycle is longer.
The content of the invention:
Present invention is primarily intended in order to overcome, existing Biochemical method oil-containing solid waste speed is not high, cycle length shows
It is defective, propose a kind of process for fixation of oil degradation flora.
Technical scheme is as follows：
A kind of process for fixation of oil degradation flora, comprises the following steps：
（1）Oil degradation flora liquid fermentation；
（2）Immobilization of the oil degradation flora on diatomite/activated carbon composite carrier.
Specifically, the oil degradation flora includes three kinds of single bacteriums, respectively pseudomonad Pseudomonus sp., bud
Spore bacillus Bacillus sp. and micrococcus luteus Micrococcus sp., according to mass ratio 1：1：1 mixing.
Specifically, step（1）The liquid fermentation includes procedure below：To the hair of the fermentation medium equipped with high-temperature sterilization
In fermentation tank, oil degradation flora liquid seeds are inoculated with according to 5-10% inoculum concentration, in 30-37 DEG C, 100-120r/min blowing airs
Cultivate 12-60h；Progress switching culture in another fermentation tank equipped with fresh fermentation medium is transferred to by 5-10% inoculum concentration, when
After number of times of transferring is more than 5, that is, obtain oil degradation bacteria mass-sending zymotic fluid.
Specifically, prepared by the oil degradation flora liquid seeds includes procedure below：The single bacterium is taken respectively, etc. quality
It is well mixed, it is inoculated according to 5-10% total inoculum concentration in fermentation medium, in 30-37 DEG C, 100-120r/min, blowing air
Cultivate 12-36h to culture medium petrochina degradation flora total quantity be 108More than CFU/mL, obtains oil degradation flora liquid strain
Son.
Specifically, the fermentation medium includes the component of following weight fraction：8-10% peptones, 3-5% beef extracts,
8-10%NaCl, the pH of the culture medium is neutrality, and liquid fermentation medium need to sterilize and use.
Specifically, step（2）The oil degradation flora immobilization includes procedure below：By step（1）In oil drop
Solution microflora fermentation liquid is mixed in equal volume with diatomite/activated carbon composite carrier after sterilizing, obtains thalline-carrier mixed liquor, so
Mixed liquor is instilled into CaCl afterwards2In solution, 12-24h is continuously stirred, it is room temperature drying, standby.
Specifically, the diatomite/activated carbon composite carrier, using process, the mass ratio of activated carbon and diatomite mixing
Example is 1:1.
Beneficial effects of the present invention：
（1）The immobilization material used is activated carbon and diatomite, wide material sources, to the immobilization role of microorganism species
Effect is good, long lifespan, cost are low, is conducive to commercialization, industrialization.
（2）Several degradation bacterias can be immobilized onto on carrier by the described method of invention simultaneously, and synergy is strong, can be achieved
The synchronous degradation of oil different component, effectively improves oil degradation speed, it is adaptable to the biodegradation of all kinds of oil-containing solid wastes.
Embodiment：
Embodiment 1：
（1）Oil degradation flora liquid fermentation
It is prepared by degradation flora liquid seeds：Take the mass mixings such as pseudomonad, bacillus and micrococcus luteus uniform respectively, press
Total inoculum concentration according to 5% is inoculated in 500mL taper triangle shaking flasks, and the liquid amount of every bottle of fermentation medium is 300mL, 30 DEG C,
100r/min blowing air culture 12h, are 10 to culture medium petrochina degradation flora total quantity8More than CFU/mL, obtains oil drop
Solve flora liquid seeds.
Wherein, fermentation medium includes the component of following weight fraction：8% peptone, 3% beef extract, 8%NaCl is described
The pH of culture medium is neutrality, and sterilizing is used.
Flora liquid fermentation：Into the fermentation tank of the fermentation medium equipped with high-temperature sterilization, it is inoculated with according to 5% inoculum concentration
Degradation flora liquid seeds, 30 DEG C, 100r/min blowing air cultures 12h；It is transferred to by 5% inoculum concentration another equipped with fresh fermentation
Switching culture is carried out in the fermentation tank of culture medium, after transferring 6 times, oil degradation bacteria is mass-sended into being fixed of zymotic fluid.
（2）Immobilization of the oil degradation flora on diatomite/activated carbon composite carrier：
By step（1）Obtained oil degradation bacteria mass-sending zymotic fluid is with diatomite/activated carbon fixation support after sterilizing etc.
Volume mixture, obtains thalline-carrier mixed liquor, mixed liquor is gently instilled into CaCl2In solution, continuously stir after 12h, room temperature is blown
It is dry, standby.
Wherein, stone diatomite/activated carbon composite carrier, using process, the mass ratio of activated carbon and diatomite mixing is
1:1。
Embodiment 2
It is prepared by degradation flora liquid seeds：Take the mass mixings such as pseudomonad, bacillus and micrococcus luteus uniform respectively, press
Total inoculum concentration according to 10% is inoculated in 500mL taper triangle shaking flasks, and the liquid amount of every bottle of fermentation medium is 300mL, 37 DEG C,
120r/min blowing air culture 36h, are 10 to culture medium petrochina degradation flora total quantity8More than CFU/mL, obtains oil drop
Solve flora liquid seeds.
Wherein, fermentation medium includes the component of following weight fraction：10% peptone, 5% beef extract, 10%NaCl, institute
It is neutrality to state the pH of culture medium, and sterilizing is used.
Flora liquid fermentation：Into the fermentation tank of the fermentation medium equipped with high-temperature sterilization, it is inoculated with according to 10% inoculum concentration
Degradation flora liquid seeds, 37 DEG C, 120r/min blowing air cultures 48h；It is transferred to by 10% inoculum concentration another equipped with fresh fermentation
Switching culture is carried out in the fermentation tank of culture medium, after transferring 6 times, oil degradation bacteria is mass-sended into being fixed of zymotic fluid.
（2）Immobilization of the oil degradation flora on diatomite/activated carbon composite carrier：
By step（1）Obtained oil degradation bacteria mass-sending zymotic fluid is with diatomite/activated carbon fixation support after sterilizing etc.
Volume mixture, obtains thalline-carrier mixed liquor, mixed liquor is gently instilled into CaCl2In solution, continuously stir after 24h, room temperature is blown
It is dry, standby.
Wherein, stone diatomite/activated carbon composite carrier, using process, the mass ratio of activated carbon and diatomite mixing is
1:1。
Embodiment 3
It is prepared by degradation flora liquid seeds：Take the mass mixings such as pseudomonad, bacillus and micrococcus luteus uniform respectively, press
Total inoculum concentration according to 8% is inoculated in 500mL taper triangle shaking flasks, and the liquid amount of every bottle of fermentation medium is 300mL, 35 DEG C,
110r/min blowing air culture 24h, are 10 to culture medium petrochina degradation flora total quantity8More than CFU/mL, obtains oil drop
Solve flora liquid seeds.
Wherein, liquid fermentation medium includes the component of following weight fraction：9% peptone, 4% beef extract, 9%NaCl,
The pH of the culture medium is neutrality, and sterilizing is used.
Flora liquid fermentation：Into the fermentation tank of the fermentation medium equipped with high-temperature sterilization, it is inoculated with according to 8% inoculum concentration
Degradation flora liquid seeds, 35 DEG C, 120r/min blowing air cultures 24h；It is transferred to by 8% inoculum concentration another equipped with fresh fermentation
Switching culture is carried out in the fermentation tank of culture medium, after transferring 6 times, oil degradation bacteria is mass-sended into being fixed of zymotic fluid.
（2）Immobilization of the oil degradation flora on diatomite/activated carbon composite carrier：
By step（1）Obtained oil degradation bacteria mass-sending zymotic fluid is with diatomite/activated carbon fixation support after sterilizing etc.
Volume mixture, obtains thalline-carrier mixed liquor, mixed liquor is gently instilled into CaCl2In solution, continuously stir after 24h, room temperature is blown
It is dry, standby.
Wherein, stone diatomite/activated carbon composite carrier, using process, the mass ratio of activated carbon and diatomite mixing is
1:1。
Analyzing of applying effects of the immobilization degradation flora in oil-based drill cuttings petrochina alkane degradation
Application example 1：
Oil degradation flora：By the immobilization of embodiment 1
Oil-based drill cuttings：Initial oil content 30000mg/kg
In the application, nutritive salt is added in oil-based drill cuttings first, makes the C in oil-based drill cuttings:N:P=100:10:1（Quality
Than）, while it is 7.0-8.0 to adjust solid waste upper strata leachate pH, by immobilization degradation flora according to weight than being applied for 0.05%
Dosage is added in oil-based drill cuttings, then sequentially add electron acceptor, surfactant, leavening agent, and water content control 20%~
30%, it is well mixed.Turn over once per 7d, after 15 days, determine oil-based drill cuttings petrochina hydrocarbon content, oil hydrocarbon content by
30000mg/kg is down to 10350mg/kg, and oily clearance is up to 65.6%.
Application example 2：
Oil degradation flora：By the immobilization of embodiment 2
Oil-based drill cuttings：Initial oil content 85000mg/kg
In the application, nutritive salt is added in oil-based drill cuttings first, makes the C in oil-based drill cuttings:N:P=100:10:1（Quality
Than）, while it is 7.0-8.0 to adjust solid waste upper strata leachate pH, by immobilization degradation flora according to weight than being applied for 0.05%
Dosage is added in oil-based drill cuttings, then sequentially add electron acceptor, surfactant, leavening agent, and water content control 20%~
30%, it is well mixed.Turn over once per 7d, after 30 days, determine oil-based drill cuttings petrochina hydrocarbon content, oil hydrocarbon content by
85000mg/kg is down to 20100mg/kg, and oily clearance is up to 76.4%.
Application example result shows that immobilized bacterium group energy is enough to carry out efficient degradation to oil-based drill cuttings petrochina hydrocarbon pollutant,
After degraded 30 days, oil content is 2%, reaches HJ607-2011《Waste mineral oil recycles pollution control technology specification》Middle drilling cuttings
Safety dumping standard.
Claims (3)
1. a kind of process for fixation of oil degradation flora, it is characterised in that comprise the following steps：
(1) oil degradation flora liquid fermentation；
(2) immobilization of the oil degradation flora on diatomite/activated carbon composite carrier；
Wherein, step (1) the oil degradation flora includes pseudomonad Pseudomonus sp., bacillus
Sp. with micrococcus luteus Micrococcus sp.；Pseudomonad Pseudomonus sp., bacillus sp. and microballoon
Bacterium Micrococcus sp., according to mass ratio 1:1:1 mixing；
The oil degradation flora liquid fermentation includes procedure below：To the fermentation tank of the fermentation medium equipped with high-temperature sterilization
In, oil degradation flora liquid seeds are inoculated with according to 5-10% inoculum concentration, in 30-37 DEG C, the training of 100-120r/min blowing airs
Support 12-60h；Progress switching culture in another fermentation tank equipped with fresh fermentation medium is transferred to by 5-10% inoculum concentration, when
After number of times of transferring is more than 5, that is, obtain oil degradation bacteria mass-sending zymotic fluid；
The oil degradation flora immobilization includes procedure below：By oil degradation bacteria mass-send zymotic fluid with sterilizing after diatomite/
Activated carbon composite carrier is mixed in equal volume, obtains the carrier mixed liquor of thalline one, mixed liquor then is instilled into CaC12In solution, even
Continuous stirring 12-24h, it is room temperature drying, standby；
The mass ratio of the stone diatomite/activated carbon composite carrier, activated carbon and diatomite mixing is 1:1.
2. the process for fixation of oil degradation flora according to claim 1, it is characterised in that the oil degradation flora
Prepared by liquid seeds include procedure below：Oil degradation flora is taken, fermentation medium is inoculated according to 5-10% total inoculum concentration
In, in 30-37 DEG C, 100-120r/min, blowing air culture 12-36h to culture medium petrochina degradation flora total quantity is
108More than CFU/mL, obtains oil degradation flora liquid seeds.
3. the process for fixation of oil degradation flora according to claim 1, it is characterised in that the fermentation medium bag
Component containing following weight fraction：8-10% peptones, 3-5% beef extracts, 8-10%NaCl, during the pH of the culture medium is
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| CN100347285C (en) * | 2005-12-21 | 2007-11-07 | 中国石油化工股份有限公司 | Liquid microbe preparation for use in petroleum pollution degradation |
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