CN104560938B - A kind of preparation method and applications of immobilizing oil degradation bacteria embedded particles - Google Patents
A kind of preparation method and applications of immobilizing oil degradation bacteria embedded particles Download PDFInfo
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- CN104560938B CN104560938B CN201310517924.4A CN201310517924A CN104560938B CN 104560938 B CN104560938 B CN 104560938B CN 201310517924 A CN201310517924 A CN 201310517924A CN 104560938 B CN104560938 B CN 104560938B
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Abstract
The present invention relates to a kind of preparation method of immobilizing oil degradation bacteria embedded particles, including：（1）35.8wt% ~ 46.5wt% water hyacinth, 14.2wt% ~ 18.5wt% wheat stalk are mixed with 35wt% ~ 50wt% walnut shell activated carbon, as assistant carrier；（2）3.0wt% ~ 7.0wt% assistant carrier is taken, access 10wt% ~ 20wt% oil degradation bacteria seed liquor is adsorbed；（3）8.0wt% ~ 10wt% polyvinyl alcohol is dissolved in 80 ~ 100 DEG C of water, adds 1.2wt% ~ 1.5wt% sodium alginates to mix, the water of surplus is added after cooling；（4）Will（2）With（3）Composition embedding system is mixed, instills CaCl2Balling-up is crosslinked in solution, then is transferred to Na2SO4Immobilization in solution, obtain immobilization embedded particles.Immobilizing oil degradation bacteria embedded particles prepared by the present invention are applied to the in-situ immobilization of oil-polluted soils, have good repairing effect to oil-polluted soils.
Description
Technical field
The present invention relates to microbial immobilized field, and in particular to the oil degradation bacteria for remedying oil-polluted soils is consolidated
Surely the preparation method and applications of embedded particles are changed.
Background technology
Oil-polluted soils are one of important pollutions of oil and petroleum chemical enterprise, and petroleum pollution therein is to human body and ring
Border has very high toxic action.The pollution of soil has been to be concerned by more and more people, and ensures the efficiency of soil remediation, recovers strong
Health production also just becomes more and more important.
The processing method of domestic and international oil-polluted soils mainly has：Burning method, landfill method, stack method and biological restoration
Deng.But it is most economical, most thorough with bioanalysis, it is the most effective means for realizing Soil ecology restoration.In order to solve at traditional biological
In reason technology field experiment free microorganism with indigenous bacterium competitiveness is weak, mithridatism pollution capacity is poor, is easily impacted by external environment
And the shortcomings of being difficult in adapt to environment, in recent years, the widely used immobilized microorganism technique of people is situated between to repair contaminated fluid
Matter（Such as waste water）With semifluid medium（Such as mud）, it have that cell density is high, microbial activity is high and stably, resistance to poison ability
The features such as strong.Therefore, nearly ten years, contaminated non-fluid medium is repaired to it（Such as soil）Research it is also increasing, but
The report for being widely used in petroleum-polluted soil reparation is also less.
The selection of fixation support and process for fixation is the critical problem of immobilized microorganism technique.Carrier property is
It is related to the key factor of immobilization technology success or failure.Fixation support mainly has organic carrier, inorganic carrier and complex carrier.Reason
The fixation support thought should possess following characteristics：It is non-toxic to microbial cell；Matrix permeability is good, and mass-transfer performance is stable；
High mechanical strength, service life length；It is cheap；High recycling rate, secondary pollution etc. is not produced after use.Traditional fixation
Change method mainly has absorption method and investment, and prepared by absorption method easy, and carrier is renewable, but the combination degree of microorganism and carrier
It is not firm enough；The bacterium amount of investment embedding is big, cell not easy to leak, and stability is good, is resistant to the infringement of noxious material, but may
Part cytoactive can be caused to lose.
CN101177677A discloses one kind and is used for polycyclic aromatic hydrocarbon（PHAs）It is prepared by the immobilization particle of contaminated soil remediation
Method, it will cultivate 12 ~ 14h seed liquor according to the ratio of weight 5 ~ 20% and coagulant liquid mixes, in 25 ~ 40 DEG C and pH nature bars
Under part, immobilization particle is made by pellet fabrication device.Coagulant liquid percent mass proportioning is：Sodium alginate 2 ~ 3%,（150 mesh）Activity
Charcoal, red brick powder, peat soil and/or flyash 0.3 ~ 0.6%, surplus are water.The invention using sodium alginate as main carrier material,
With activated carbon, red brick powder, peat soil and/or flyash etc. for assistant carrier, traditional sodium alginate embedded immobilization is improved
It technique, can greatly improve the mechanical strength, elasticity and sustained release performance of immobilization particle.But this method is directly entered to thalline
Row embedding, influences the activity of thalline, and cannot guarantee that adsorbance of the carrier to thalline, prepared flyash immobilization
Particle is up to 54.1% and 45.8% to P in soil YR and BaP clearance, and other immobilization particles are low to PYR and BaP clearances
In 50% and 40%.
The content of the invention
In view of the shortcomings of the prior art, the present invention provide a kind of immobilizing oil degradation bacteria embedded particles preparation method and
It is applied.Immobilizing oil degradation bacteria embedded particles prepared by the inventive method can be used for the in-situ immobilization of oil-polluted soils,
There is good repairing effect to oil-polluted soils, be it is a kind of can be with the environment-friendly products of popularization and application.
The preparation method of immobilizing oil degradation bacteria embedded particles of the present invention, including herein below：（1）By 35.8wt% ~
46.5wt% water hyacinth, 14.2wt% ~ 18.5wt% wheat stalk crush sieving, the walnut chitin activity with 35wt% ~ 50wt%
Charcoal mixes, as assistant carrier；（2）The assistant carrier for accounting for embedding system 3.0wt% ~ 7.0wt% is taken, access 10wt% ~ 20wt%'s
Oil degradation bacteria seed liquor, is adsorbed；（3）Embedding system 8.0wt% ~ 10wt% polyvinyl alcohol (PVA) stirring and dissolving will be accounted for
In 80 ~ 100 DEG C of water, the sodium alginate for adding 1.2wt% ~ 1.5wt% is mixed, and the water of surplus is added after cooling；（4）Will（2）With
（3）Composition embedding system is mixed, instills CaCl2Balling-up is crosslinked in solution, then is transferred to Na2SO4Immobilization in solution, obtain oil
Degradation bacteria immobilization embedded particles.
In the present invention, step（1）Described assistant carrier is preferably pre-processed, the water calabash that will be mixed in proportion first
Reed, wheat stalk and walnut shell activated carbon mixture 12 ~ 24h of distilled water immersion, pH7.0 ~ 7.2 are adjusted, it is miscellaneous to filter off upper strata
Matter, then 50 DEG C of the mixture dry 24h, 121 DEG C of high pressure steam sterilization 20min that will be filtered out.
In the present invention, step（1）The full carbon content of described water hyacinth is 220g/kg ~ 260g/kg, total nitrogen content is
38.5g/kg ~ 45.5g/kg, content of tatal phosphorus are the g/kg of 9 g/kg ~ 10.8.The full carbon content of wheat stalk be 28.1g/kg ~
35.6g/kg, total nitrogen content are 32.9g/kg ~ 46.8g/kg, content of tatal phosphorus is the g/kg of 0.64 g/kg ~ 0.83.Walnut chitin
Activated carbon is Powdered Activated Carbon, and its specific surface area is 960m2/ g, iodine sorption value 820mg/g.Walnut shell activated carbon is use
ZnCl2Prepared by activation method, specific preparation method is：Weigh a certain amount of walnut shell to pulverize and sieve and be impregnated in mass concentration
50% ZnCl2In solution, solid-liquid ratio 2:3, stirring stands 24 h after 2 hours, is put into after drying in tubular heater, in 300
℃、N2Atmosphere under carbonize 80min, gained carbonizes sample again in 50%ZnCl220min is impregnated in solution, through filtering, drying it is rearmounted
40 min are activated at 500 DEG C in Muffle furnace, activation products are boiled into 20 min with the hydrochloric acid of mass fraction 20% after cooling, then
Products therefrom is washed to neutrality with deionized water, is placed in 120 DEG C of drying in drying box.Walnut shell activated carbon surface area is big,
Adsorption capacity is strong, can effectively adsorb mycelium and resolvase, and its porous can also increase the diffusion speed of product and substrate
Degree, diffusional resistance is reduced, increase the permeable structures inside microballoon.Water hyacinth and wheat stalk contained fibrous matter and battalion in itself
Material is supported, contributes to charcoal absorption thalline, ensures that the bacterium amount of absorption meets to require, while the nutrition need of thalline can also be met
Ask.
In the present invention, step（2）The preparation of described oil degradation bacteria seed liquor uses beef extract-peptone Liquid Culture
Base, the quantity for exponential phase and the point of interface of stationary phase, ensureing oil degradation bacteria is cultivated under the conditions of 120r/min, 30 DEG C
Reach 1 × 1010~5×1010Individual/mL.By oil degradation bacteria culture to exponential phase and the point of interface of stationary phase, it is ensured that
The most highly active and stability of bacterium.
In the present invention, step（2）The absorption of described oil degradation bacteria is cultivated under the conditions of 120 ~ 160r/min, 30 DEG C
16 ~ 24h is adsorbed, oil degradation bacteria absorption is ensured that the bacterium amount of absorption reaches 9 × 10 on assistant carrier9~4.6×1010Individual/
mL。
In the present invention, step（4）The CaCl of use2The concentration of solution is 1wt% ~ 5wt%, Na2SO4The concentration of solution is
10wt%.After being crosslinked balling-up, by microballoon taking-up brine, Na is put into2SO4In solution, continue reaction 2 under stirring condition
~ 4h, brine is then used again, finally obtained immobilizing oil degradation bacteria embedded particles.
The application of immobilizing oil degradation bacteria embedded particles of the present invention, it is specially：First the oil degradation bacteria of preparation is fixed
Change embedded particles aseptically to access in proliferated culture medium by 10wt% ~ 20wt% ratio, 120 ~ 160r/min, 30 DEG C
Under the conditions of incubated 24h, then filter culture medium, 50 ~ 100g/kg petroleum-contaminated soils spread by every square metre in a manner of sowing
0.1 ~ 1kg immobilizing oil degradation bacteria embedded particles, soil 30wt% ~ 40wt% moisture is kept, and to 0 ~ 30cm of top layer
Soil is turned over, and is allowed to be well mixed with immobilizing oil degradation bacteria embedded particles, and ensures the good aeration of soil.
Compared with prior art, the inventive method has following outstanding feature：
1st, with natural inorganic carrier（Walnut shell activated carbon）With natural organic carrier（Water hyacinth and wheat stalk）
Mixture as assistant carrier, immobilizing oil degradation bacteria embedded particles are prepared using the method for absorption-embedding combination, can be with
Oil degradation bacteria is set to be preferentially adsorbed on assistant carrier, can be with microballoon using assistant carrier as attachment carrier and nutriment
More preferable growth and breeding, mycelium is secreted, it is smaller to the activity influence of immobilized bacterium than directly embedding, it can preferably adapt to the external world
The impact of environment.
2nd, used carrier of the present invention to biological nontoxic, mass-transfer performance is good, adsorption capacity is big, mechanical strength is big, acid and alkali-resistance, the longevity
Life length, can also provide nutriment for the growth and breeding of thalline, overcome exclusive use natural inorganic carrier and require supplementation with outside
Carry out nutrient source and the shortcomings that natural organic supports fix bacterium amount deficiency is used alone.
3rd, water hyacinth and wheat stalk are supplied to the nitrogen of microorganism, phosphorus mass ratio to have close to the N/P ratio in cell component
Beneficial to raising microbial degradation ability.Meanwhile on the one hand the N/P ratio can meet the needs of microbial growth is to nitrogen P elements,
The biological prosthetic demand to nitrogen P elements of oil-polluted soils can also be met by slowly discharging, without extra addition nitrogen phosphorus
Nutrient source.
4th, wheat stalk contains substantial amounts of cellulose and hemicellulose material, has good water retention property, micro- improving
While permeability, offer nitrogen and phosphorus source, increase activated carbon inhale bacterium amount inside ball, the watering in repair process can also be reduced
The frequency and watering amount, meet during bacterial remediation needed for moisture.
5th, immobilizing oil degradation bacteria embedded particles have obvious repairing effect to oil-polluted soils, to 50g/kg ~
100g/kg petroleum-contaminated soils, in 0.5 ~ 1kg/m of rate of charge2Under conditions of soil, repair time 40d, petroleum pollution clearance can
Up to 55% ~ 75%, specific ionization bacterium is higher by 21% ~ 33%, and wherein saturated hydrocarbons removal effect is best, up to 65% ~ 85%.
Embodiment
The present invention is further described below by embodiment, but and is not so limited the present invention.In the present invention,
Wt% is mass fraction.
Embodiment 1
1st, the preparation of assistant carrier
Water hyacinth and wheat stalk are dried, crushed, 80 mesh sieves is crossed, is then mixed with walnut shell activated carbon, its reclaimed water
Cucurbit 46.5wt%, wheat stalk 18.5wt%, walnut shell activated carbon 35wt%.Mixture distilled water immersion 24h, regulation
PH7.0, filter off the impurity and supernatant of upper strata floating, the dry 24h of 50 DEG C of mixture that will be filtered out, 121 DEG C of high pressure steam sterilizations
20min, it is standby to make assistant carrier.
The full carbon content of water hyacinth in assistant carrier is 250g/kg, total nitrogen content 43.8g/kg, content of tatal phosphorus are
10.2g/kg.The full carbon content of wheat stalk is 32.6g/kg, total nitrogen content is 40.8 g/kg, content of tatal phosphorus 0.76g/kg.
Walnut shell activated carbon is Powdered Activated Carbon, and its specific surface area is 960m2/ g, iodine sorption value 820mg/g.In assistant carrier
Carbon N/P ratio is 25:5.7:1, close to the N/P ratio in cell component, microbial growth on the one hand can be met to nitrogen P elements
Demand, while the biological prosthetic demand to nitrogen P elements of oil-polluted soils can also be met by slowly discharging, without
Extra addition nitrogen and phosphorus source.
Walnut shell activated carbon is to use ZnCl2Prepared by activation method, specific preparation method is：Weigh a certain amount of crushed
The walnut shell of sieve is impregnated in the ZnCl of mass concentration 50%2In solution, solid-liquid ratio 2:3, stirring stands 24 h after 2 hours, after drying
It is put into tubular heater, in 300 DEG C, N2Atmosphere under carbonize 80min, gained carbonizes sample again in 50%ZnCl2Soaked in solution
Stain 20min, through filtering, dry after be placed in Muffle furnace at 500 DEG C and activate 40 min, by activation products quality point after cooling
The hydrochloric acid solution of number 20% boils 20 min, then is washed products therefrom to neutrality with deionized water, is placed in drying box in 120
DEG C drying.
2nd, the preparation of seed liquor
Oil degradation bacteria is inoculated in beef extract-peptone fluid nutrient medium, cultivated under the conditions of 120r/min, 30 DEG C
To the exponential phase of oil degradation bacteria and the point of interface of stationary phase, the seed liquor of oil degradation bacteria is made, ensures in seed liquor
The quantity of oil degradation bacteria reaches 1 × 1010Individual/mL.The formula of beef extract-peptone fluid nutrient medium is：NaCl 10g/L, egg
White peptone 10g/L, beef extract 5g/L and certain volume distilled water are well mixed, and pH7.0 is adjusted with NaOH or hydrochloric acid, 121 DEG C
20 min of vapor sterilizing.
3rd, the preparation of immobilization embedded particles
1. culture is taken to exponential phase and the 20wt% of the point of interface of stationary phase oil degradation bacteria seed liquor and 7wt%
Assistant carrier mixes, and 120r/min, adsorbs 16h under the conditions of 30 DEG C, oil degradation bacteria absorption is ensured absorption on assistant carrier
Bacterium amount reach 9 × 109Individual/mL.
2. by 9wt% polyvinyl alcohol stirring and dissolving in 80 ~ 100 DEG C of water, add 1.35wt% sodium alginates and mix, cooling
To after 40 DEG C, the water of surplus is added, is then well mixed with the assistant carrier after 1. middle absorption thalline, forms embedding system.Most
Afterwards the CaCl that concentration is 1wt% is instilled with syringe2Balling-up is crosslinked in solution, microballoon is taken out after 30min, washed with physiology salt
After washing 3 times, 10wt% Na is transferred to2SO4In solution, continue to react 3h under stirring condition, then with brine, final system
Obtain immobilizing oil degradation bacteria embedded particles.
4th, the application of immobilization embedded particles
The mass concentration that 20wt% is pressed under the immobilizing oil degradation bacteria embedded particles aseptic condition of preparation is first accessed into propagation
In culture medium, 120r/min, incubated 24h under the conditions of 30 DEG C, immobilization oil degradation bacteria is activated.Culture medium is filtered, to broadcast
The mode of kind spreads 0.5kg immobilizing oil degradation bacteria embedded particles to 100g/kg petroleum-contaminated soils by every square metre, keeps soil
35wt% moisture, and top layer 0 ~ 30cm soil is turned over, under conditions of repair time 40d, petroleum pollution removes
Rate is up to 70.3%, and specific ionization bacterium is higher by 32.2%, and wherein saturated hydrocarbons removal effect is best, up to 79.1%.
Described proliferation culture medium formula is：The g/L of glucose 20, the g/L of yeast extract 3, the g/L of peptone 2, ammonium nitrate 1
G/L, MgSO4•7H27.0 ~ 7.5,115 DEG C of 30 m in of sterilizing of the g/L of O 0.2 g/L, KCl 0.2, regulation pH.
Embodiment 2
One of do not exist together and be with embodiment 1：In the preprocessing process of assistant carrier, the proportioning of each material is：Water hyacinth
43wt%, wheat stalk 17wt%, walnut shell activated carbon 40wt%.Two not existed together are：Prepare oil degradation bacteria seed liquor
When, ensure that seed liquor petrochina degradation bacteria quantity reaches 3 × 1010Individual/mL.Three not existed together are：Immobilizing oil degradation bacteria
When prepared by embedded particles, the quality for adding polyvinyl alcohol is 10wt%, sodium alginate 1.5wt%, CaCl2The concentration of solution is 3wt%
In, in 10wt% Na2SO4Reaction time in solution is 2h.
Other conditions are identical, and under conditions of repair time 40d, petroleum pollution clearance is gone up to 64.7%, wherein saturated hydrocarbons
Except rate is up to 73%.
Embodiment 3
One of do not exist together and be with embodiment 1：In the preprocessing process of assistant carrier, the proportioning of each material is：Water hyacinth
35.8wt%, wheat stalk 14.2wt%, walnut shell activated carbon 50wt%.Two not existed together are：Prepare oil degradation bacteria seed
During liquid, ensure that the quantity of seed liquor petrochina degradation bacteria reaches 5 × 1010Individual/mL.Three not existed together are：Oil degradation bacteria is consolidated
When changing embedded particles surely and preparing, the assistant carrier of addition is 3wt%, 120r/min, cultivates 24h under the conditions of 30 DEG C.Add PVA's
Quality is 8wt%, sodium alginate 1.2wt%, CaCl2The concentration of solution is 5wt%, in 10wt% Na2SO4Reaction time in solution
For 2h.Four not existed together are：Petroleum-contaminated soil oil-containing 50g/kg, the immobilizing oil degradation bacteria embedded particles amount of being sprinkled into are 1kg/
m2。
Other conditions are identical, and under conditions of repair time 40d, petroleum pollution clearance is gone up to 59.3%, wherein saturated hydrocarbons
Except rate is up to 64.6%.
Comparative example 1
One of do not exist together and be with embodiment 1, it is 960m that assistant carrier, which is all specific surface area,2/ g, iodine sorption value are
820mg/g powdered walnut shell activated carbon.
Other conditions are identical, and under conditions of repair time 40d, petroleum pollution clearance is gone up to 48.9%, wherein saturated hydrocarbons
Except rate is 55%.
Comparative example 2
One of not existing together and be with embodiment 1, assistant carrier is all water hyacinth and wheat stalk, wherein, water hyacinth accounts for
71.6wt%, wheat stalk 28.4wt%.Two not existed together are：When prepared by immobilizing oil degradation bacteria embedded particles, fail to protect
The bacterium amount adsorbed on card assistant carrier reaches 9 × 109Individual/mL.
Other conditions are identical, and under conditions of repair time 40d, petroleum pollution clearance is gone up to 51.6%, wherein saturated hydrocarbons
Except rate is 58.5%.
Comparative example 3
One of do not exist together and be with embodiment 1, it is 960m that assistant carrier, which is all specific surface area,2/ g, iodine sorption value are
820mg/g powdered walnut shell activated carbon.The two of difference are：Soil watering is being turned over using immobilization particle
While applied fertilizer, it is 25 to ensure carbon N/P ratio:5.7:1.
Other conditions are identical, and under conditions of repair time 40d, petroleum pollution clearance removes up to 72%, wherein saturated hydrocarbons
Rate is up to 80.2%.
Comparative example 4
Difference from Example 1 is：When prepared by immobilizing oil degradation bacteria embedded particles, directly 9wt% PVA are stirred
Mix and be dissolved in 80 ~ 100 DEG C of water, add 1.35wt% sodium alginate solns and be well mixed, add the mixing of 7wt% assistant carriers
Uniformly, after being cooled to 40 DEG C or so, the water of surplus is added, then with cultivating to exponential phase and the point of interface of stationary phase
20wt% oil degradation bacteria seed liquor is mixed together uniformly, forms embedding system.Then it is 1wt%'s to instill concentration with syringe
CaCl2Solution crosslinking balling-up, framboid is taken out after 30min, after brine 3 times, be transferred to 10wt% Na2SO4It is molten
In liquid, continue to react 3h under stirring condition, then with brine, immobilizing oil degradation bacteria embedded particles are made.
By obtained immobilizing oil degradation bacteria embedded particles in a manner of sowing by 0.5kg/m2It is sprinkled into 100g/kg greasy dirts
In soil, soil 40wt% moisture is kept, and top layer 0 ~ 30cm soil is turned over, repairs 40d, petroleum pollution
Clearance is 58.65%, and specific ionization bacterium is higher by 20.55%, and wherein saturated hydrocarbons clearance is up to 63%.
Claims (7)
1. a kind of preparation method of immobilizing oil degradation bacteria embedded particles, including herein below：（1）By 35.8wt% ~
46.5wt% water hyacinth, 14.2wt% ~ 18.5wt% wheat stalk crush sieving, the walnut chitin activity with 35wt% ~ 50wt%
Charcoal mixes, and as assistant carrier, described walnut shell activated carbon is Powdered Activated Carbon, specific surface area 960m2/ g, iodine absorption
It is worth for 820mg/g；（2）The assistant carrier for accounting for embedding system 3.0wt% ~ 7.0wt% is taken, accesses 10wt% ~ 20wt% oil degradation
Bacterium seed liquor, is adsorbed；The preparation of described oil degradation bacteria seed liquor uses beef extract-peptone fluid nutrient medium,
120r/min, culture to exponential phase and the point of interface of stationary phase, ensures that the quantity of oil degradation bacteria reaches 1 under the conditions of 30 DEG C
×1010~5×1010Individual/mL；The absorption of described oil degradation bacteria is that absorption is cultivated under the conditions of 120 ~ 160r/min, 30 DEG C
16 ~ 24h, oil degradation bacteria absorption is set to ensure that the bacterium amount of absorption reaches 9 × 10 on assistant carrier9~4.6×1010Individual/mL；
（3）Embedding system 8.0wt% ~ 10wt% polyvinyl alcohol stirring and dissolving will be accounted in 80 ~ 100 DEG C of water, add 1.2wt% ~ 1.5wt%
Sodium alginate mix, after cooling add surplus water；（4）Will（2）With（3）Composition embedding system is mixed, instills CaCl2Solution
Middle crosslinking balling-up, then it is transferred to Na2SO4Immobilization in solution, obtain immobilizing oil degradation bacteria embedded particles.
2. in accordance with the method for claim 1, it is characterised in that：Step（1）Described assistant carrier is pre-processed, and is first used
12 ~ 24h of distilled water immersion, pH7.0 ~ 7.2 are adjusted, filter off upper strata impurity, then 50 DEG C of the mixture dry 24h that will be filtered out, 121 DEG C
High pressure steam sterilization 20min.
3. in accordance with the method for claim 1, it is characterised in that：Step（1）The full carbon content of described water hyacinth is 220g/kg
~ 260g/kg, total nitrogen content are 38.5g/kg ~ 45.5g/kg, and content of tatal phosphorus is 9 g/kg ~ 10.8g/kg.
4. in accordance with the method for claim 1, it is characterised in that：Step（1）The full carbon content of described wheat stalk is
28.1g/kg ~ 35.6g/kg, total nitrogen content are 32.9g/kg ~ 46.8g/kg, and content of tatal phosphorus is 0.64 g/kg ~ 0.83g/kg.
5. in accordance with the method for claim 1, it is characterised in that：Step（4）CaCl2The concentration of solution is 1wt% ~ 5wt%,
Na2SO4The concentration of solution is 10wt%.
6. in accordance with the method for claim 1, it is characterised in that：Step（4）After being crosslinked balling-up, by microballoon taking-up physiology salt
Water washing；Place into Na2SO4In solution, 2 ~ 4h of stirring reaction, then use brine.
7. the application of the immobilizing oil degradation bacteria embedded particles prepared according to any described method of claim 1 to 6, it is special
Sign is：The immobilizing oil degradation bacteria embedded particles of preparation are aseptically accessed in 10wt% ~ 20wt% ratio and increased
Grow in culture medium, 120 ~ 160r/min, incubated 24h under the conditions of 30 DEG C, then filter culture medium, in a manner of sowing pair
50 ~ 100g/kg petroleum-contaminated soils spread 0.1 ~ 1kg immobilizing oil degradation bacteria embedded particles by every square metre, keep soil 30wt% ~
40wt% moisture, and top layer 0 ~ 30cm soil is turned over.
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