CN104403982A - Preparation method of oil displacement microbe high-density concentrated microbial inoculant by fermentation and freeze-drying - Google Patents
Preparation method of oil displacement microbe high-density concentrated microbial inoculant by fermentation and freeze-drying Download PDFInfo
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- CN104403982A CN104403982A CN201410802821.7A CN201410802821A CN104403982A CN 104403982 A CN104403982 A CN 104403982A CN 201410802821 A CN201410802821 A CN 201410802821A CN 104403982 A CN104403982 A CN 104403982A
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- 238000006073 displacement reaction Methods 0.000 title claims abstract description 23
- 238000004108 freeze drying Methods 0.000 title claims abstract description 23
- 238000000855 fermentation Methods 0.000 title claims abstract description 18
- 230000004151 fermentation Effects 0.000 title claims abstract description 18
- 230000000813 microbial effect Effects 0.000 title abstract description 18
- 238000002360 preparation method Methods 0.000 title abstract description 4
- 239000002054 inoculum Substances 0.000 title abstract 8
- 239000007788 liquid Substances 0.000 claims abstract description 31
- 238000000034 method Methods 0.000 claims abstract description 21
- 238000005138 cryopreservation Methods 0.000 claims abstract description 9
- 239000000843 powder Substances 0.000 claims abstract description 6
- 238000002156 mixing Methods 0.000 claims abstract description 5
- 235000019198 oils Nutrition 0.000 claims description 35
- 241000894006 Bacteria Species 0.000 claims description 24
- 239000002068 microbial inoculum Substances 0.000 claims description 21
- 241000122230 Acinetobacter junii Species 0.000 claims description 10
- 230000006872 improvement Effects 0.000 claims description 9
- 239000002609 medium Substances 0.000 claims description 9
- 235000015097 nutrients Nutrition 0.000 claims description 9
- 229960005486 vaccine Drugs 0.000 claims description 9
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 230000001954 sterilising effect Effects 0.000 claims description 8
- 230000004083 survival effect Effects 0.000 claims description 7
- 239000008399 tap water Substances 0.000 claims description 7
- 235000020679 tap water Nutrition 0.000 claims description 7
- 239000008176 lyophilized powder Substances 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 4
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 4
- 235000013379 molasses Nutrition 0.000 claims description 4
- 235000013923 monosodium glutamate Nutrition 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 235000020183 skimmed milk Nutrition 0.000 claims description 4
- 229940073490 sodium glutamate Drugs 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 235000020238 sunflower seed Nutrition 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 3
- 239000008101 lactose Substances 0.000 claims description 3
- 238000005457 optimization Methods 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 238000012512 characterization method Methods 0.000 claims 1
- 239000012141 concentrate Substances 0.000 claims 1
- 239000001963 growth medium Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 8
- 238000004321 preservation Methods 0.000 abstract description 4
- 239000003223 protective agent Substances 0.000 abstract description 4
- 238000005406 washing Methods 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract 1
- 238000011049 filling Methods 0.000 abstract 1
- 239000003921 oil Substances 0.000 description 29
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 11
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 238000007710 freezing Methods 0.000 description 3
- 230000008014 freezing Effects 0.000 description 3
- 235000011187 glycerol Nutrition 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000011169 microbiological contamination Methods 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- 230000009257 reactivity Effects 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 230000007423 decrease Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 238000012807 shake-flask culturing Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 239000006137 Luria-Bertani broth Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000010779 crude oil Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000005966 endogenous activation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 208000018875 hypoxemia Diseases 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000003716 rejuvenation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C09K8/00—Compositions for drilling of boreholes or wells; Compositions for treating boreholes or wells, e.g. for completion or for remedial operations
- C09K8/58—Compositions for enhanced recovery methods for obtaining hydrocarbons, i.e. for improving the mobility of the oil, e.g. displacing fluids
- C09K8/582—Compositions for enhanced recovery methods for obtaining hydrocarbons, i.e. for improving the mobility of the oil, e.g. displacing fluids characterised by the use of bacteria
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Abstract
The invention relates to a preparation method of an oil displacement microbe high-density concentrated microbial inoculant by fermentation and freeze-drying, belonging to the technical field of preparation of a microbe oil displacement high-density concentrated microbial inoculant. The method comprises the following steps: a. preparing a high-concentration fermentation liquid by a high-density culture technique; b. washing the fermentation liquid with sterile normal saline 2-3 times, and carrying out centrifugal concentration; c. adding a right amount of freeze-drying protective agent, and uniformly mixing; d. uniformly filling the microbial liquid into a cryopreservation tube; and e. freeze-drying in a vacuum freeze drier to obtain the microbial inoculant. The method can be used for preparing the higher-concentration microbial inoculant and the freeze-dried powder thereof, and is simple and effective to operate; the strain activity is high; the concentration of the concentrated microbial inoculant can reach 10<9>-10<10> cfu/mL; the microbe content of the freeze-dried microbial powder is 10<10>-10<11> cfu/mL; and the microbial inoculant has the advantage of longer preservation time, is convenient to carry and reduces the transportation cost for the oil displacement field microbial inoculant.
Description
Technical field
The invention belongs to microbial oil displacement Concentrated Compact microbial inoculum preparing technical field, be specifically related to a kind of Concentrated Compact bacteria fermentation and freeze-drying process of oil displacement microbes.
Background technology
Domestic and international at present can only the crude oil of extraction subsurface deposit 30% ~ 40% with conventional oil recovery technique, improving oil recovery is one of key issue of petroleum industry research both at home and abroad at present.In recent years since, utilized Microbial-flooding Technology, namely utilize microbe metabolite tensio-active agent, organic acid, microbial profile control, biogas to increase oil and interfacial effect etc., achieve certain effect, oil production rate increases to some extent.Under laboratory condition, the microbial oil displacement effect of artificial culture is better, but under field condition, due to the imprecision of technical process, the shortage of source of clean water, the deficiency of heating and thermal insulation equipment, cause being easy to microbiological contamination, death in fermentation process, thalline fermentation can not reach quantity required, affects oil displacement efficiency.
Current microbial oil displacement mainly contains two kinds of technology, and a kind of method is inject oil reservoir after adopting the oil displacement microbes of screening to cultivate, and another kind of method is injected nutrition for utilization and activated indigenous microorganism.Oil reservoir is a kind of particular surroundings, mineral composition wherein and character, porosity and permeability, reservoir pressure, the temperature of fluid, pH, salinity, oil property, residual oil saturation etc., all on microorganism growth generation significantly impact, itself can contain some endogenous microbes again in oil reservoir simultaneously, after injection inoculating microbe or nutrition, also can there are some changes, and interact with inoculating microbe.Therefore be high density, the high reactivity that external source injection or endogenous activation all need to ensure aimed strain, ensure that oil displacement microbes occupies advantage.
The high-activity fermented liquid that field factories is produced is the important factor affecting oil displacement efficiency, and the oil displacement microbes bacterial classification of high reactivity, high density is an important indicator of restriction plant produced.The routine techniques fermentation that current oil field adopts, because various condition limits, can only by the concentration of thalline by original 10
3~ 10
4cfu/mL brings up to 10
6~ 10
7cfu/mL, and be easy to microbiological contamination.Because needs fermented liquid repeatedly circulates injection oil reservoir, fermented liquid adopts the method for glycerine fluid-tight to carry out preservation usually, because storage conditions is severe, be difficult to reach the condition such as low temperature, anoxic and reduce the metabolic condition of thalline, cause the microbial activity in fermented liquid lower, thalline mortality ratio is high, and it is longer that fermented liquid fermentation period is prepared by factory, and thalline microbiological contamination rate is high.Under laboratory condition, the bacterium of conventional glycerine preservation Oil-displacement Bacteria is dense only has 10
7~ 10
8cfu/mL, bacterium powder concentration are 10
8~ 10
9cfu/g, thalline survival rate only have 1.4% ~ 9.8%.
The present invention, by high-density culture technology, not only increases fermented liquid concentration, and improves the productive rate of final Fungal biodiversity and target product, reduce bio-reactor volume, reduces facility investment, shortens fermentation period, reduces water power and uses.The fermented liquid of high density carries out concentrated freeze-dried by the present invention simultaneously, not only simple to operation, and microbial inoculum is easy to carry about with one, and reduces on-the-spot transportation cost.Can the seed liquor of acquisition high density rapidly and efficiently when field factories is produced, decrease fermentation period, the object suppressing other microbial contaminations can be reached simultaneously, be conducive to improving oil recovery.
Summary of the invention
For the shortcoming and defect that existing microbial oil displacement is produced, the invention provides a kind of method of preparing of high density, high reactivity microbial inoculum that can be permanently effective.
In order to achieve the above object, experimental program is as follows:
1. the slant strains being stored in refrigerator is inoculated on solid LB media and activated for 2 ~ 3 generations.
2. by 30 ~ 37 DEG C, 140 ~ 160r/min fermentation culture, 20 ~ 24h in the colony inoculation on solid LB flat board to the liquid nutrient medium of improvement.
3. fermented liquid is put into centrifuge tube 4000 ~ 8000r/min, centrifugal 10 ~ 15min, abandon supernatant, add the lyophilized vaccine of optimization in proportion, fully mix.
4. the microbial inoculum of mixing is put into cryopreservation tube or Freeze Drying Equipment.
5. the microbial inoculum of mixing is distinguished pre-freeze 20min ~ 40min, 1h ~ 2h, 4h ~ 6h at 0 ~ 4 DEG C ,-20 DEG C ~ 30 DEG C ,-60 DEG C ~ 80 DEG C refrigerators successively, after using Freeze Drying Equipment freeze-drying, concentrated freeze-dried microbial inoculum can be obtained.
The liquid nutrient medium of described Acinetobacter junii improvement consists of: sunflower seed oil 1% ~ 10% (v/v), molasses 0.1% ~ 0.5% (m/v), sodium-chlor 3 ~ 5g, peptone 6 ~ 10g, yeast extract paste 3 ~ 5g, 1L is settled to, pH5.0 ~ 5.5 with tap water; Pseudomonas aeruginosa improvement liquid nutrient medium consists of: sunflower seed oil 2% ~ 8% (v/v), molasses 0.5% ~ 3% (m/v), sodium-chlor 3 ~ 5g, peptone 6 ~ 10g, extractum carnis 3 ~ 8g, 1L is settled to, pH7.0 ~ 8.5 with tap water.
The described the suitableeest lyophilized vaccine of Acinetobacter junii is combined as (m/v): skim-milk 1% ~ 10%, Sodium Glutamate 1% ~ 10%, sucrose 1% ~ 10%; The suitableeest lyophilized vaccine of Pseudomonas aeruginosa is (m/v): lactose 10% ~ 15%.121 DEG C of sterilizing 15min.
The invention has the advantages that:
The present invention is by carrying out single factor test and orthogonal design optimization to different fermentations substratum composition, lyophilized vaccine composition; obtain the suitableeest fermention medium composition, the lyophilized vaccine composition of oil displacement microbes; vacuum freeze is utilized to reach the conditions such as the hypoxemia of culture presevation, low water, low temperature; in Oil Field test; by microbial inoculum concentration technique; substantially prolongs the preservation time of bacterial classification; ensure that the relative stability of spawn activity simultaneously; rejuvenation of spawn is rapid; activity is recovered very soon, has saved displacement of reservoir oil microbial inoculum fermentation time.Concentrated Compact technology makes that bacterial classification bacterium is dense reaches 10
10~ 10
11cfu/mL, thalline lyophilized powder bacteria containing amount reaches 10
11~ 10
12cfu/g, survival rate is up to more than 74.1%.It is 10 that conventional glycerine is preserved bacterium dense
7~ 10
8cfu/mL, bacterium powder lyophilized powder bacteria containing amount 10
8~ 10
9cfu/g, thalline survival rate only has 1.4% ~ 9.8%.The preparation of freeze-drying microbial inoculum decreases the industrial cost of bacterium liquid, ensure that microbial inoculum biomass is with active, can also suppress living contaminants simultaneously, improve microbial oil displacement effect.
Embodiment
Embodiment 1
1. prepare 50mL Acinetobacter junii improvement liquid nutrient medium, 121 DEG C of sterilizing 15min, the Acinetobacter junii bacterial classification of access activation, 30 DEG C, 140r/min shake-flask culture 24h.
2. prepare freezing protective agent: skim-milk 1g, Sodium Glutamate 1g, sucrose 1g, with tap water constant volume to 100mL, 121 DEG C of sterilizing 15min, for subsequent use.
3. get fermented liquid 1mL bacterium liquid respectively, draw fermented liquid 10mL respectively with pipettor, the centrifugal 10min of 6000r/min; with after stroke-physiological saline solution washing, the same terms is centrifugal again; add Acinetobacter junii respectively and optimize protective material 1mL, fully mix, obtaining bacterium dense is 2.1 × 10
10the frozen storing liquid of cfu/mL.Last often pipe 0.2mL is dispensed in 2mL cryopreservation tube, and by cryopreservation tube successively after 4 DEG C ,-20 DEG C ,-80 DEG C refrigerators respectively pre-freeze 30min, 1h, 4h, use Freeze Drying Equipment freeze-drying 20h, obtain concentrated freeze-dried microbial inoculum, gained bacterium is dense is 1.7 × 10
10cfu/mL, lyophilized powder bacteria containing amount is 2.3 × 10
10cfu/g, freeze-drying thalline survival rate is 81.0%.
Embodiment 2
1. prepare 1.5L Pseudomonas aeruginosa improvement liquid nutrient medium, put into 3L fermentor tank, 121 DEG C of sterilizing 15min, the Pseudomonas aeruginosa bacterial classification of access activation, 36 DEG C, 160r/min fermentation culture 24h.
2. prepare freezing protective agent: lactose 15g, with tap water constant volume to 100mL, 121 DEG C of sterilizing 15min, for subsequent use.
3. draw fermented liquid 100mL respectively, the centrifugal 15min of 7000r/min, with after stroke-physiological saline solution washing, the same terms is centrifugal again, add Pseudomonas aeruginosa respectively and optimize protective material 1mL, fully mix, obtaining bacterium dense is 1.8 × 10
11the frozen storing liquid of cfu/mL. last often pipe 0.2mL is dispensed in 2mL cryopreservation tube, and by cryopreservation tube successively after 4 DEG C ,-20 DEG C ,-80 DEG C refrigerators respectively pre-freeze 30min, 2h, 4h, use Freeze Drying Equipment freeze-drying 22h, obtain concentrated freeze-dried microbial inoculum, gained bacterium is dense is 1.4 × 10
11cfu/mL, lyophilized powder bacteria containing amount is 3.6 × 10
12cfu/g, freeze-drying thalline survival rate is 77.8%.
Embodiment 3
1. prepare 50mL Acinetobacter junii improvement liquid nutrient medium, 121 DEG C of sterilizing 15min, the Acinetobacter junii bacterial classification of access activation, 37 DEG C, 160r/min shake-flask culture 24h.
2. prepare freezing protective agent: skim-milk 5g, Sodium Glutamate 10g, sucrose 5g, with tap water constant volume to 100mL, 121 DEG C of sterilizing 15min, for subsequent use.
3. draw fermented liquid 10mL respectively, the centrifugal 15min of 8000r/min, with after stroke-physiological saline solution washing, the same terms is centrifugal again, add respectively and optimize protective material 1mL, after fully mixing, obtaining bacterium dense is 2.1 × 10
10the frozen storing liquid of cfu/mL.Last often pipe 0.2mL is dispensed in 2mL cryopreservation tube, and by cryopreservation tube successively after 4 DEG C ,-20 DEG C ,-80 DEG C refrigerators respectively pre-freeze 30min, 1h, 4h, use Freeze Drying Equipment freeze-drying 24h, obtaining concentrated freeze-dried microbial inoculum gained bacterium dense is 1.5 × 10
10cfu/mL, lyophilized powder bacteria containing amount is 3.4 × 10
11cfu/g, freeze-drying thalline survival rate is 71.4%.
Claims (8)
1. the Concentrated Compact bacteria fermentation of oil displacement microbes and a freeze-drying process, its characterization step is: a, the oil displacement microbes slant strains being stored in refrigerator to be inoculated on solid LB plate culture medium and to activate for 2 ~ 3 generations; B, by colony inoculation after the activation on solid LB flat board to improvement liquid nutrient medium in fermentation 20 ~ 24h, obtain high-density oil displacement microbes fermented liquid; C, fermented liquid is centrifugal after, abandon supernatant, obtain Concentrated Compact microbial inoculum, add the lyophilized vaccine of optimization in proportion, mix; D, the microbial inoculum of mixing is evenly dispensed in cryopreservation tube or Freeze Drying Equipment; E, microbial inoculum respectively after pre-freeze 20min ~ 40min, 1h ~ 2h, 4h ~ 6h, after using Freeze Drying Equipment freeze-drying, can be obtained concentrated microbial inoculum lyophilized powder successively under 0 ~ 4 DEG C ,-20 DEG C ~ 30 DEG C ,-60 DEG C ~ 80 DEG C conditions.
2. method according to claim 1, is characterized in that: oil displacement microbes is Acinetobacter junii and Pseudomonas aeruginosa.
3. method according to claim 1, it is characterized in that: in step b, the liquid nutrient medium of Acinetobacter junii improvement consists of: sunflower seed oil 1% ~ 10% (v/v), molasses 0.1% ~ 0.5% (m/v), sodium-chlor 3 ~ 5g, peptone 6 ~ 10g, yeast extract paste 3 ~ 5g, 1L is settled to, pH5.0 ~ 5.5 with tap water; Pseudomonas aeruginosa improvement liquid nutrient medium consists of: sunflower seed oil 2% ~ 8% (v/v), molasses 0.5% ~ 3% (m/v), sodium-chlor 3 ~ 5g, peptone 6 ~ 10g, extractum carnis 3 ~ 8g, 1L is settled to, pH7.0 ~ 8.5 with tap water; Culture condition is: 30 DEG C ~ 37 DEG C, 140 ~ 180r/min.
4. method according to claim 1, is characterized in that: in step c, centrifugal condition is temperature 0 ~ 8 DEG C, rotating speed 4000 ~ 8000r/min, 10 ~ 15min.
5. method according to claim 1, is characterized in that: it is 10 that step c middle-high density concentrates microbial inoculum bacterium dense
10~ 10
11cfu/mL.
6. method according to claim 1, is characterized in that: in step c, the suitableeest lyophilized vaccine of Acinetobacter junii is combined as (m/v): skim-milk 1% ~ 10%, Sodium Glutamate 1% ~ 10%, sucrose 1% ~ 10%; The suitableeest lyophilized vaccine of Pseudomonas aeruginosa is (m/v): lactose 10% ~ 15%, 121 DEG C of sterilizing 15min.
7. method according to claim 1, is characterized in that: the ratio of the lyophilized vaccine in step c and concentrated microbial inoculum is 1: 1 ~ 1: 100.
8. method according to claim 1, is characterized in that: in step e, microbial inoculum dry powder water content is: 2% ~ 8%, and microbial inoculum dry powder bacteria containing amount is 10
11~ 10
12cfu/g, survival rate is to more than 71.4%.
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Cited By (5)
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| CN106520616A (en) * | 2016-11-07 | 2017-03-22 | 中国石油大学(北京) | Acinetobacter junii for producing bio-surfactant and application of acinetobacter junii |
| CN108102980A (en) * | 2018-02-07 | 2018-06-01 | 沈阳建筑大学 | A kind of method that salt tolerant denitrification compound bacteria agent freezes the preparation and its processing high-salt wastewater of bacterium powder |
| WO2022142381A1 (en) * | 2020-12-31 | 2022-07-07 | 东南大学 | Method for preparing mineralized microbial powder |
| CN117363332A (en) * | 2023-12-04 | 2024-01-09 | 胜利星科石油技术开发(山东)有限公司 | Microbial oil extraction and displacement system and preparation method and application thereof |
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2014
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106244459A (en) * | 2016-08-31 | 2016-12-21 | 沈阳化工研究院有限公司 | A kind of pseudomonas dry powder and preparation method thereof |
| CN106520616A (en) * | 2016-11-07 | 2017-03-22 | 中国石油大学(北京) | Acinetobacter junii for producing bio-surfactant and application of acinetobacter junii |
| CN106520616B (en) * | 2016-11-07 | 2019-08-06 | 中国石油大学(北京) | The Acinetobacter junii of biosurfactant production and its application |
| CN108102980A (en) * | 2018-02-07 | 2018-06-01 | 沈阳建筑大学 | A kind of method that salt tolerant denitrification compound bacteria agent freezes the preparation and its processing high-salt wastewater of bacterium powder |
| CN108102980B (en) * | 2018-02-07 | 2021-07-06 | 沈阳建筑大学 | Preparation of salt-tolerant denitrification composite microbial inoculum freeze-dried bacterial powder and method for treating high-salinity wastewater by using same |
| WO2022142381A1 (en) * | 2020-12-31 | 2022-07-07 | 东南大学 | Method for preparing mineralized microbial powder |
| CN117363332A (en) * | 2023-12-04 | 2024-01-09 | 胜利星科石油技术开发(山东)有限公司 | Microbial oil extraction and displacement system and preparation method and application thereof |
| CN117363332B (en) * | 2023-12-04 | 2024-02-13 | 胜利星科石油技术开发(山东)有限公司 | Microbial oil extraction and displacement system and preparation method and application thereof |
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