CN109880756A - A kind of preparation method of oil degradation microbial inoculum - Google Patents
A kind of preparation method of oil degradation microbial inoculum Download PDFInfo
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Abstract
The present invention relates to a kind of preparation methods of oil degradation microbial inoculum, belong to oil pollution recovery technique field.Pomelo peel and active carbon are carried out pyrolysis processing by the present invention, as the carrier of bacterium solution, also have hole more containing a large amount of cellulose pomelo peel in the white flock layer inside pomelo peel, the big feature in aperture, pomelo peel has compact institutional framework, network structure is unobvious, has certain absorption to oil product, the pomelo peel tridimensional network after being pyrolyzed becomes apparent, big aperture staggeredly communicates, be conducive to fast absorbing oil, increase obviously before the relatively pyrolysis of aperture, be conducive to the raising of oil absorption；Artificial carrier can preferably match, to obtain more preferably immobilization effect and repairing performance because having the advantages that controllable specific surface area, pore-size distribution and mass-transfer performance are good with specific microorganism and complicated pollutant；Charcoal has stronger adsorption capacity to petroleum, realizes enrichment-degradation integration of pollutant, peomotes soil remediation effect.
Description
Technical field
The present invention relates to a kind of preparation methods of oil degradation microbial inoculum, belong to oil pollution recovery technique field.
Background technique
With the increasingly increase of oil extraction, in exploitation, transport, storage and accident leakage, petroleum largely into
Enter environment, seriously polluted soil, underground water, river and ocean.It is a Xiang Qing using the soil of microorganism remediation oil pollution
The Low investment of clean environment, high benefit, the emerging technology convenient for applying, development potentiality is big.Petroleum is in production, processing and use process
In, due to upset operation, accident and maintenance etc., the spilling and discharge of petroleum hydrocarbon are had, causes petroleum to environment
Pollution, repair pollution it is extremely urgent.Microorganism remediation technology is a kind of efficient, safe, inexpensive oil pollution reparation skill
Art, but free microorganism influences in actually pollution environment vulnerable to extraneous factor, and degradation is difficult to give full play to.
In order to eliminate the pollution of petroleum, remedying oil-polluted to experienced quite long technological innovation and research, people are to stone
The restorative procedure of oily pollution has done style, mainly there is physical method, chemical method and bioanalysis.
Physical method is remedying oil-polluted using physical method, comprising: gas floatation process, membrane separation process, absorption method, heat treating process
(incineration method) soil replacement method, isolation method etc..Absorption method is one of most common physical method, is using with porous solids
Matter achievees the effect that one or more substances move to solid phase surface from liquid phase.Active carbon is generally physical absorption carrier
Selection.Physical method further includes chemical flocculation, chemical oxidization method, electrochemical process.
It is biological prosthetic to contaminated soil progress, it is primarily referred to as utilizing specific biological (plant, microorganism or protozoan)
By the growth and metabolic process of biology itself, removing, degrade or convert harmful substance is innocuous substance, reduces dustiness, extensive
Multiple good environment and ecological effect.Common biological renovation method mainly has several: (1) microorganism remediation, microorganism
It repairs mainly using petroleum as nutriment needed for growth, petroleum is degraded.Microorganism remediation divides biology in situ to repair
Multiple and Ex situ bioremediation.(2) phytoremediation, phytoremediation technology are root and the intracorporal enzyme of plant using plant to pollution
The method administered of environment.(3) plant-microorganism joint repair, it is actual it is biological prosthetic in, plant-microorganism connection
Zoarium system can promote fast degradation, the mineralising of petroleum.
The bioremediation technology producing cost of oil-polluted soils is low, does not generate secondary pollution, be one have it is wide before
The new and high technology of scape.Therefore, how to become researchers at home and abroad using biological prosthetic oil-polluted soils to try to explore to solve
Important topic.Exist in itself in soil it is some can with the indigenous microorganism of degraded oil, they be people obtain it is biological prosthetic
The major microorganisms source of soil.
Summary of the invention
The technical problems to be solved by the invention: it for the problem that the existing oil degradation microbial inoculum holding time is limited, provides
A kind of preparation method of oil degradation microbial inoculum.
In order to solve the above technical problems, the technical solution adopted by the present invention is that:
(1) fresh grapefruit skin is crushed, is placed in the baking oven that temperature is 90~100 DEG C and dries to constant weight to get broken shaddock
Skin mixes broken pomelo peel and active carbon, carries out pyrolysis processing, is cooled to room temperature, and grinds and crosses 30~50 meshes to get mixed
Close object；
(2) oil degradation bacteria is accessed into seed culture medium, solid culture is added to get seed liquor in culture processing in seed liquor
Base, culture processing is to get bacterium solution；
(3) mixture and bacterium solution are mixed, carries out vacuumize process, filtered up to filter residue；By acid fiber by polylactic and ionic liquid
Mixing, is stirred, is cooled to room temperature to get mixed solution, filter residue is added in mixed solution, stir and be spray-dried
Processing, is cooled to room temperature to get oil degradation microbial inoculum.
Broken pomelo peel and active carbon are mixed described in step (1), carry out pyrolysis treatment step are as follows: in mass ratio 1: 1
Broken pomelo peel and active carbon are mixed, 3~4h is pyrolyzed in the Muffle furnace that temperature is 300~400 DEG C.
The preparation step of oil degradation bacteria described in step (2) are as follows: in mass ratio 1: 1 by bacillus subtilis and false unit cell
Bacterium mixes to get oil degradation bacteria.
The component and preparation step of seed culture medium described in step (2) are as follows: beef extract 5g, peptone 10g, sodium chloride
5g, distilled water 1000mL, adjusting pH value is 7.0, and sterilize 20min at being 121 DEG C in temperature.
The component and preparation step of solid medium described in step (2) are as follows: sucrose 10g, turf 10g, beef extract 6g, ferment
Female powder 1.5g, distilled water 1000mL, sterilize 20min at being 121 DEG C in temperature.
Oil degradation bacteria is accessed into seed culture medium described in step (2), cultivates processing step are as follows: connect oil degradation bacteria
Enter seed culture medium, inoculum concentration 5% is 25~30 DEG C in temperature, and revolving speed is that 12~20h is cultivated under 160~200r/min.
Solid medium is added described in step (2) in seed liquor, cultivates processing step are as follows: in mass ratio 1: 1 is planting
Solid medium is added in sub- liquid, stirs evenly, is cultivated 2~3 days at being 25~30 DEG C in temperature.
Mixture and bacterium solution are mixed described in step (3), carry out vacuumize process step are as follows: in mass ratio 1: 20 will mix
Object and bacterium solution mixing are closed, is placed in obturator, 2~4h is continuously vacuumized, vacuum environment is kept to stand 1~2 day.
Acid fiber by polylactic and ionic liquid are mixed described in step (3), are stirred step are as follows: in mass ratio 1:
10 are uniformly mixed acid fiber by polylactic and ionic liquid, stir 2~3h at being 70~75 DEG C in temperature.
Filter residue is added described in step (3) in mixed solution, stirs simultaneously spray drying treatment step are as follows: in mass ratio 5:
1 is added filter residue in mixed solution, low whipping speed be stirred evenly under 500~600r/min, and inlet air temperature be 110~
120 DEG C, outlet temperature is that 5~10min is spray-dried at 49~53 DEG C.
The present invention is compared with other methods, and advantageous effects are:
(1) pomelo peel and active carbon are carried out pyrolysis processing by the present invention, the white flock as the carrier of bacterium solution, inside pomelo peel
Contain a large amount of cellulose, hemicellulose in layer, pomelo peel also has hole more, the big feature in aperture, and pomelo peel has compact
Institutional framework, inner wall is rough, more folds, and hole is more, and aperture is larger, and network structure is unobvious, has to oil product certain
Absorption, the pomelo peel tridimensional network after being pyrolyzed become apparent, and big aperture staggeredly communicates, and are conducive to fast absorbing oil, aperture
It is obvious compared with increasing before pyrolysis, be conducive to the raising of oil absorption；Artificial carrier because have controllable specific surface area, pore-size distribution with
And mass-transfer performance it is good the advantages that, can preferably be matched with specific microorganism and complicated pollutant, to more preferably be fixed
Change effect and repairing performance；Charcoal has stronger adsorption capacity to petroleum, prepares oil degradation microbial inoculum by carrier of charcoal
After adding soil, can promote pollutant from soil to fixation support migrate, make fixation support Sync enrichment microorganism and
Pollutant increases contact of the microorganism with pollutant, realizes enrichment-degradation integration of pollutant, peomotes soil and repair
Multiple effect；
(2) mode that liquid and solid-state phase of the invention combine cultivates the oil degradation microbial inoculum comprising 2 kinds of bacteriums, beef
Cream, peptone are then seeded into the solid medium of sucrose and turf and cultivate as liquid culture medium culture primary seed, system
Standby high efficient petroleum degrading bacteria agent a kind of out；Organic matter and humic acid are rich in turf, water conservation ventilation is that excellent soil improvement produces
Product, pseudomonad energy secreting surfactant rhamnolipid, evenly spread to petroleum in water phase, and bacillus subtilis has height
The oil degradation performance of effect, two plants of bacterium are used in combination, and can dramatically increase the contact surface of petroleum and bacterial strain, improve petroleum degradation rate；
(3) present invention prepares a kind of oil degradation microbial inoculum that the holding time is long using physisorphtion and investment, with pyrolysis
Pomelo peel and active carbon afterwards is carrier, and mixed bacteria is adsorbed in matrix surface or bore area, will using acid fiber by polylactic
Bacterium solution is limited in the small grid of gel, while matrix can be allowed to penetrate into and diffused out with product, so that oil degradation microbial inoculum
Holding time extends；
(4) present invention prepares oil degradation microbial inoculum using spray drying process, and material solution is dispersed the liquid that nebulizes by atomizer
Drop, carries out heat exchange under the action of dried medium (hot wind), evaporates solvent in misty liquid droplets rapidly, obtain powder
Drying process.Drop temperature is relatively low in the drying process, thus the drying particularly suitable for heat sensitive material (biological products),
And quality of finished product good.
Specific embodiment
Fresh grapefruit skin is crushed, is placed in the baking oven that temperature is 90~100 DEG C and dries to constant weight to get broken shaddock
Skin, in mass ratio 1: 1 mixes broken pomelo peel and active carbon, and 3~4h is pyrolyzed in the Muffle furnace that temperature is 300~400 DEG C,
It is cooled to room temperature, grinds and crosses 30~50 meshes to get mixture；In mass ratio 1: 1 by bacillus subtilis and pseudomonad
Mixing is to get oil degradation bacteria；Seed culture medium are as follows: beef extract 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL are adjusted
Saving pH value is 7.0, and sterilize 20min at being 121 DEG C in temperature；Solid medium are as follows: sucrose 10g, turf 10g, beef extract 6g, ferment
Female powder 1.5g, distilled water 1000mL, sterilize 20min at being 121 DEG C in temperature；Oil degradation bacteria is accessed into seed culture medium, is connect
Kind amount is 5%, is 25~30 DEG C in temperature, and revolving speed is that 12~20h of culture is under 160~200r/min to get seed liquor, by quality
Solid medium is added in seed liquor than 1: 1, stirs evenly, cultivates 2~3 days at being 25~30 DEG C in temperature to get bacterium solution；
In mass ratio 1: 20 mixes mixture and bacterium solution, is placed in obturator, continuously vacuumizes 2~4h, keeps vacuum environment quiet
After setting 1~2 day, filter up to filter residue；In mass ratio 1: 10 is uniformly mixed acid fiber by polylactic and ionic liquid, is 70 in temperature
2~3h is stirred at~75 DEG C, is cooled to room temperature and filter residue is added in mixed solution to get mixed solution, in mass ratio 5: 1,
Mixing speed is 110~120 DEG C in inlet air temperature to stir evenly under 500~600r/min, and outlet temperature is 49~53 DEG C
5~10min of lower spray drying.It is cooled to room temperature to get oil degradation microbial inoculum.
Example 1
Fresh grapefruit skin is crushed, is placed in baking oven at a temperature of 90 °C and dries to constant weight to get broken pomelo peel, will be crushed
Pomelo peel and active carbon mixing, carry out pyrolysis processing, are cooled to room temperature, and grind and cross 30 meshes to get mixture；Petroleum is dropped
Solve bacterium and access seed culture medium, solid medium is added in seed liquor to get seed liquor in culture processing, culture processing to get
Bacterium solution；Mixture and bacterium solution are mixed, vacuumize process is carried out, is filtered up to filter residue；Acid fiber by polylactic and ionic liquid are mixed
It closes, is stirred, is cooled to room temperature to get mixed solution, filter residue is added in mixed solution, stir and be spray-dried place
Reason, is cooled to room temperature to get oil degradation microbial inoculum.Broken pomelo peel and active carbon are mixed, carry out pyrolysis treatment step are as follows: press
Mass ratio 1: 1 mixes broken pomelo peel and active carbon, is pyrolyzed 3h in the Muffle furnace that temperature is 300 DEG C.Oil degradation bacteria
Preparation step are as follows: in mass ratio 1: 1 mixes bacillus subtilis and pseudomonad to get oil degradation bacteria.Seed culture medium
Component and preparation step are as follows: beef extract 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, adjust pH value be 7.0,
Temperature is the 20min that sterilizes at 121 DEG C.The component and preparation step of solid medium are as follows: sucrose 10g, turf 10g, beef extract 6g,
Yeast powder 1.5g, distilled water 1000mL, sterilize 20min at being 121 DEG C in temperature.Oil degradation bacteria is accessed into seed culture medium,
Cultivate processing step are as follows: oil degradation bacteria is accessed into seed culture medium, inoculum concentration 5% is 25 DEG C in temperature, revolving speed 160r/
12h is cultivated under min.Solid medium is added in seed liquor, cultivates processing step are as follows: in mass ratio 1: 1 adds in seed liquor
Enter solid medium, stir evenly, is cultivated 2 days at being 25 DEG C in temperature.Mixture and bacterium solution are mixed, vacuumize process is carried out
Step are as follows: in mass ratio 1: 20 mixes mixture and bacterium solution, is placed in obturator, continuously vacuumizes 2h, keeps vacuum ring
Border stands 1 day.Acid fiber by polylactic and ionic liquid are mixed, step is stirred are as follows: in mass ratio 1: 10 by polylactic acid
Fiber and ionic liquid are uniformly mixed, and stir 2h at being 70 DEG C in temperature.Filter residue is added in mixed solution, stir and does by spraying
Dry processing step are as follows: in mass ratio 5: 1 are added filter residue in mixed solution, and low whipping speed is to stir evenly under 500r/min,
And inlet air temperature be 110 DEG C, outlet temperature be 49 DEG C at be spray-dried 5min.
Example 2
Fresh grapefruit skin is crushed, is placed in the baking oven that temperature is 93 DEG C and dries to constant weight to get broken pomelo peel, will be crushed
Pomelo peel and active carbon mixing, carry out pyrolysis processing, are cooled to room temperature, and grind and cross 39 meshes to get mixture；Petroleum is dropped
Solve bacterium and access seed culture medium, solid medium is added in seed liquor to get seed liquor in culture processing, culture processing to get
Bacterium solution；Mixture and bacterium solution are mixed, vacuumize process is carried out, is filtered up to filter residue；Acid fiber by polylactic and ionic liquid are mixed
It closes, is stirred, is cooled to room temperature to get mixed solution, filter residue is added in mixed solution, stir and be spray-dried place
Reason, is cooled to room temperature to get oil degradation microbial inoculum.Broken pomelo peel and active carbon are mixed, carry out pyrolysis treatment step are as follows: press
Mass ratio 1: 1 mixes broken pomelo peel and active carbon, is pyrolyzed 3h in the Muffle furnace that temperature is 330 DEG C.Oil degradation bacteria
Preparation step are as follows: in mass ratio 1: 1 mixes bacillus subtilis and pseudomonad to get oil degradation bacteria.Seed culture medium
Component and preparation step are as follows: beef extract 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, adjust pH value be 7.0,
Temperature is the 20min that sterilizes at 121 DEG C.The component and preparation step of solid medium are as follows: sucrose 10g, turf 10g, beef extract 6g,
Yeast powder 1.5g, distilled water 1000mL, sterilize 20min at being 121 DEG C in temperature.Oil degradation bacteria is accessed into seed culture medium,
Cultivate processing step are as follows: oil degradation bacteria is accessed into seed culture medium, inoculum concentration 5% is 26 DEG C in temperature, revolving speed 170r/
14h is cultivated under min.Solid medium is added in seed liquor, cultivates processing step are as follows: in mass ratio 1: 1 adds in seed liquor
Enter solid medium, stir evenly, is cultivated 2 days at being 26 DEG C in temperature.Mixture and bacterium solution are mixed, vacuumize process is carried out
Step are as follows: in mass ratio 1: 20 mixes mixture and bacterium solution, is placed in obturator, continuously vacuumizes 3h, keeps vacuum ring
Border stands 1 day.Acid fiber by polylactic and ionic liquid are mixed, step is stirred are as follows: in mass ratio 1: 10 by polylactic acid
Fiber and ionic liquid are uniformly mixed, and stir 2h at being 72 DEG C in temperature.Filter residue is added in mixed solution, stir and does by spraying
Dry processing step are as follows: in mass ratio 5: 1 are added filter residue in mixed solution, and low whipping speed is to stir evenly under 530r/min,
And inlet air temperature be 113 DEG C, outlet temperature be 50 DEG C at be spray-dried 6min.
Example 3
Fresh grapefruit skin is crushed, is placed in the baking oven that temperature is 96 DEG C and dries to constant weight to get broken pomelo peel, will be crushed
Pomelo peel and active carbon mixing, carry out pyrolysis processing, are cooled to room temperature, and grind and cross 45 meshes to get mixture；Petroleum is dropped
Solve bacterium and access seed culture medium, solid medium is added in seed liquor to get seed liquor in culture processing, culture processing to get
Bacterium solution；Mixture and bacterium solution are mixed, vacuumize process is carried out, is filtered up to filter residue；Acid fiber by polylactic and ionic liquid are mixed
It closes, is stirred, is cooled to room temperature to get mixed solution, filter residue is added in mixed solution, stir and be spray-dried place
Reason, is cooled to room temperature to get oil degradation microbial inoculum.Broken pomelo peel and active carbon are mixed, carry out pyrolysis treatment step are as follows: press
Mass ratio 1: 1 mixes broken pomelo peel and active carbon, is pyrolyzed 3h in the Muffle furnace that temperature is 360 DEG C.Oil degradation bacteria
Preparation step are as follows: in mass ratio 1: 1 mixes bacillus subtilis and pseudomonad to get oil degradation bacteria.Seed culture medium
Component and preparation step are as follows: beef extract 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, adjust pH value be 7.0,
Temperature is the 20min that sterilizes at 121 DEG C.The component and preparation step of solid medium are as follows: sucrose 10g, turf 10g, beef extract 6g,
Yeast powder 1.5g, distilled water 1000mL, sterilize 20min at being 121 DEG C in temperature.Oil degradation bacteria is accessed into seed culture medium,
Cultivate processing step are as follows: oil degradation bacteria is accessed into seed culture medium, inoculum concentration 5% is 29 DEG C in temperature, revolving speed 180r/
18h is cultivated under min.Solid medium is added in seed liquor, cultivates processing step are as follows: in mass ratio 1: 1 adds in seed liquor
Enter solid medium, stir evenly, is cultivated 3 days at being 29 DEG C in temperature.Mixture and bacterium solution are mixed, vacuumize process is carried out
Step are as follows: in mass ratio 1: 20 mixes mixture and bacterium solution, is placed in obturator, continuously vacuumizes 4h, keeps vacuum ring
Border stands 2 days.Acid fiber by polylactic and ionic liquid are mixed, step is stirred are as follows: in mass ratio 1: 10 by polylactic acid
Fiber and ionic liquid are uniformly mixed, and stir 3h at being 74 DEG C in temperature.Filter residue is added in mixed solution, stir and does by spraying
Dry processing step are as follows: in mass ratio 5: 1 are added filter residue in mixed solution, and low whipping speed is to stir evenly under 560r/min,
And inlet air temperature be 116 DEG C, outlet temperature be 52 DEG C at be spray-dried 8min.
Example 4
Fresh grapefruit skin is crushed, is placed in the baking oven that temperature is 100 DEG C and dries to constant weight to get broken pomelo peel, will break
Broken pomelo peel and active carbon mixing, carry out pyrolysis processing, are cooled to room temperature, and grind and cross 50 meshes to get mixture；By petroleum
Degradation bacteria accesses seed culture medium, and solid medium is added to get seed liquor in culture processing in seed liquor, and culture is handled, i.e.,
Obtain bacterium solution；Mixture and bacterium solution are mixed, vacuumize process is carried out, is filtered up to filter residue；By acid fiber by polylactic and ionic liquid
Mixing, is stirred, is cooled to room temperature to get mixed solution, filter residue is added in mixed solution, stir and be spray-dried
Processing, is cooled to room temperature to get oil degradation microbial inoculum.Broken pomelo peel and active carbon are mixed, pyrolysis treatment step is carried out are as follows:
In mass ratio 1: 1 mixes broken pomelo peel and active carbon, is pyrolyzed 4h in the Muffle furnace that temperature is 400 DEG C.Oil degradation bacteria
Preparation step are as follows: in mass ratio 1: 1 mixes bacillus subtilis and pseudomonad to get oil degradation bacteria.Seed culture
The component and preparation step of base are as follows: beef extract 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, adjusting pH value is 7.0,
Sterilize 20min at being 121 DEG C in temperature.The component and preparation step of solid medium are as follows: sucrose 10g, turf 10g, beef extract
6g, yeast powder 1.5g, distilled water 1000mL, sterilize 20min at being 121 DEG C in temperature.Oil degradation bacteria is accessed into seed culture
Base cultivates processing step are as follows: oil degradation bacteria is accessed seed culture medium, inoculum concentration 5% is 30 DEG C in temperature, and revolving speed is
20h is cultivated under 200r/min.Solid medium is added in seed liquor, cultivates processing step are as follows: in mass ratio 1: 1 in seed liquor
Middle addition solid medium, stirs evenly, and cultivates 3 days at being 30 DEG C in temperature.Mixture and bacterium solution are mixed, vacuumized
Processing step are as follows: in mass ratio 1: 20 mixes mixture and bacterium solution, is placed in obturator, continuously vacuumizes 4h, keeps true
Altitude stands 2 days.Acid fiber by polylactic and ionic liquid are mixed, are stirred step are as follows: in mass ratio 1: 10 will gather
Acid fiber and ionic liquid are uniformly mixed, and stir 3h at being 75 DEG C in temperature.Filter residue is added in mixed solution, stir and sprays
Mist is dried step are as follows: in mass ratio 5: 1 are added filter residue in mixed solution, and low whipping speed is to stir under 600r/min
It is even, and be 120 DEG C in inlet air temperature, outlet temperature is to be spray-dried 10min at 53 DEG C.
Reference examples: the oil degradation microbial inoculum of Dalian company production.
The oil degradation microbial inoculum that example and reference examples are prepared is detected, specific detection is as follows:
Petroleum removal rate: it by the microbial inoculum inoculum concentration of 0.5g, is added in crude oil culture medium and is cultivated in 160rpm, 30 DEG C of shaking tables, survey
Fixed week degradation rate.If 2 is parallel, petroleum removal rate is measured.
Specific test result such as table 1.
1 performance characterization contrast table of table
| Detection project | Example 1 | Example 2 | Example 3 | Example 4 | Reference examples |
| Removal rate/% | 50.7 | 49.8 | 49.5 | 50.3 | 16.8 |
As shown in Table 1, oil degradation microbial inoculum prepared by the present invention has good deoiling effect.
Claims (10)
1. a kind of preparation method of oil degradation microbial inoculum, it is characterised in that specific preparation step are as follows:
(1) fresh grapefruit skin is crushed, is placed in the baking oven that temperature is 90~100 DEG C and dries to constant weight to get broken shaddock
Skin mixes broken pomelo peel and active carbon, carries out pyrolysis processing, is cooled to room temperature, and grinds and crosses 30~50 meshes to get mixed
Close object；
(2) oil degradation bacteria is accessed into seed culture medium, solid culture is added to get seed liquor in culture processing in seed liquor
Base, culture processing is to get bacterium solution；
(3) mixture and bacterium solution are mixed, carries out vacuumize process, filtered up to filter residue；By acid fiber by polylactic and ionic liquid
Mixing, is stirred, is cooled to room temperature to get mixed solution, filter residue is added in mixed solution, stir and be spray-dried
Processing, is cooled to room temperature to get oil degradation microbial inoculum.
2. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (1)
Broken pomelo peel and active carbon are mixed, carry out pyrolysis treatment step are as follows: in mass ratio 1: 1 mixes broken pomelo peel and active carbon
It closes, 3~4h is pyrolyzed in the Muffle furnace that temperature is 300~400 DEG C.
3. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (2)
The preparation step of oil degradation bacteria are as follows: in mass ratio 1: 1 mixes bacillus subtilis and pseudomonad to get oil degradation
Bacterium.
4. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (2)
The component and preparation step of seed culture medium are as follows: beef extract 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL adjust pH
Value is 7.0, and sterilize 20min at being 121 DEG C in temperature.
5. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (2)
The component and preparation step of solid medium are as follows: sucrose 10g, turf 10g, beef extract 6g, yeast powder 1.5g, distilled water
1000mL, sterilize 20min at being 121 DEG C in temperature.
6. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (2)
Oil degradation bacteria is accessed into seed culture medium, cultivates processing step are as follows: oil degradation bacteria is accessed into seed culture medium, inoculum concentration is
5%, it is 25~30 DEG C in temperature, revolving speed is that 12~20h is cultivated under 160~200r/min.
7. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (2)
Solid medium is added in seed liquor, cultivates processing step are as follows: in mass ratio 1: 1 is added solid medium in seed liquor,
It stirs evenly, is cultivated 2~3 days at being 25~30 DEG C in temperature.
8. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (3)
Mixture and bacterium solution are mixed, vacuumize process step is carried out are as follows: in mass ratio 1: 20 mixes mixture and bacterium solution, is placed in close
It closes in device, continuously vacuumizes 2~4h, vacuum environment is kept to stand 1~2 day.
9. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (3)
Acid fiber by polylactic and ionic liquid are mixed, step is stirred are as follows: in mass ratio 1: 10 by acid fiber by polylactic and ion
Liquid is uniformly mixed, and stirs 2~3h at being 70~75 DEG C in temperature.
10. a kind of preparation method of oil degradation microbial inoculum according to claim 1, it is characterised in that: described in step (3)
Filter residue is added in mixed solution, stirs simultaneously spray drying treatment step are as follows: in mass ratio 5: 1 are added filter in mixed solution
Slag, low whipping speed are 110~120 DEG C in inlet air temperature to stir evenly under 500~600r/min, outlet temperature 49
5~10min is spray-dried at~53 DEG C.
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| CN111906141A (en) * | 2020-07-14 | 2020-11-10 | 西南石油大学 | Soil remediation method based on biochar immobilized degrading bacteria |
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| CN111906141A (en) * | 2020-07-14 | 2020-11-10 | 西南石油大学 | Soil remediation method based on biochar immobilized degrading bacteria |
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