CN113980950A - Preparation method of ammonia nitrogen degrading bacteria immobilized pellet - Google Patents
Preparation method of ammonia nitrogen degrading bacteria immobilized pellet Download PDFInfo
- Publication number
- CN113980950A CN113980950A CN202111319297.4A CN202111319297A CN113980950A CN 113980950 A CN113980950 A CN 113980950A CN 202111319297 A CN202111319297 A CN 202111319297A CN 113980950 A CN113980950 A CN 113980950A
- Authority
- CN
- China
- Prior art keywords
- ammonia nitrogen
- fermentation
- wall material
- mass concentration
- ochrobactrum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 title claims abstract description 50
- 241000894006 Bacteria Species 0.000 title claims abstract description 28
- 230000000593 degrading effect Effects 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 239000008188 pellet Substances 0.000 title description 10
- 239000000463 material Substances 0.000 claims abstract description 31
- 239000000725 suspension Substances 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 14
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000001768 carboxy methyl cellulose Substances 0.000 claims abstract description 14
- 239000000661 sodium alginate Substances 0.000 claims abstract description 14
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 14
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 14
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 claims abstract description 14
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 claims abstract description 14
- 238000003756 stirring Methods 0.000 claims abstract description 11
- 241000588843 Ochrobactrum Species 0.000 claims abstract description 10
- 239000011324 bead Substances 0.000 claims abstract description 10
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 9
- 238000012258 culturing Methods 0.000 claims abstract description 7
- 230000003213 activating effect Effects 0.000 claims abstract description 4
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium chloride Substances Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 claims abstract description 4
- 238000000855 fermentation Methods 0.000 claims description 32
- 230000004151 fermentation Effects 0.000 claims description 32
- 230000001580 bacterial effect Effects 0.000 claims description 29
- 230000004913 activation Effects 0.000 claims description 23
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 21
- 239000002002 slurry Substances 0.000 claims description 19
- 239000001963 growth medium Substances 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 239000011259 mixed solution Substances 0.000 claims description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 241000588814 Ochrobactrum anthropi Species 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 239000007836 KH2PO4 Substances 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 5
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000005995 Aluminium silicate Substances 0.000 claims description 4
- 229910021536 Zeolite Inorganic materials 0.000 claims description 4
- 235000012211 aluminium silicate Nutrition 0.000 claims description 4
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims description 4
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 claims description 4
- 239000010457 zeolite Substances 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 3
- ZDQYSKICYIVCPN-UHFFFAOYSA-L sodium succinate (anhydrous) Chemical compound [Na+].[Na+].[O-]C(=O)CCC([O-])=O ZDQYSKICYIVCPN-UHFFFAOYSA-L 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 238000011068 loading method Methods 0.000 claims description 2
- 239000002609 medium Substances 0.000 claims description 2
- 239000002351 wastewater Substances 0.000 abstract description 16
- 244000005700 microbiome Species 0.000 abstract description 5
- 241001052560 Thallis Species 0.000 abstract description 4
- 239000008223 sterile water Substances 0.000 abstract description 4
- 239000011159 matrix material Substances 0.000 abstract description 2
- 230000004083 survival effect Effects 0.000 abstract description 2
- 239000011162 core material Substances 0.000 abstract 1
- 230000015556 catabolic process Effects 0.000 description 9
- 238000006731 degradation reaction Methods 0.000 description 9
- 239000010802 sludge Substances 0.000 description 7
- 239000012533 medium component Substances 0.000 description 6
- 238000005406 washing Methods 0.000 description 4
- 239000000306 component Substances 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 238000000053 physical method Methods 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 229940074404 sodium succinate Drugs 0.000 description 2
- 238000004065 wastewater treatment Methods 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000010779 crude oil Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000008235 industrial water Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000005504 petroleum refining Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- PVGBHEUCHKGFQP-UHFFFAOYSA-N sodium;n-[5-amino-2-(4-aminophenyl)sulfonylphenyl]sulfonylacetamide Chemical compound [Na+].CC(=O)NS(=O)(=O)C1=CC(N)=CC=C1S(=O)(=O)C1=CC=C(N)C=C1 PVGBHEUCHKGFQP-UHFFFAOYSA-N 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Dispersion Chemistry (AREA)
- Molecular Biology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Inorganic Chemistry (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
The invention discloses a preparation method of immobilized beads of ammonia nitrogen degrading bacteria. Firstly, preparing a strain suspension by activating strains and fermenting and culturing ochrobactrum; taking a suspension of the ochrobactrum sp as a core material, and taking sodium alginate, sodium carboxymethylcellulose and an ammonia nitrogen adsorbing materialMixing and stirring as wall material, and adding AlCl3And sterile water as a cross-linking agent to prepare the immobilized beads. Sodium alginate, sodium carboxymethylcellulose and ammonia nitrogen adsorbing materials form a space network, the structure is compact so as to fully protect thalli, a survival matrix is provided for microorganisms, the loss of thalli is reduced, and the obtained material has good stability and reusability and can efficiently treat ammonia nitrogen petrochemical wastewater.
Description
Technical Field
The invention belongs to the field of wastewater treatment technology and bioengineering. In particular to a preparation method of immobilized beads of ammonia nitrogen degrading bacteria.
Technical Field
China has a serious problem of water resource shortage, and in recent years, with the rapid development of industry and the continuous improvement of the living standard of people, the condition that water resources are polluted is continuously worsened. At present, the petrochemical industry in China develops rapidly, the quantity of processed crude oil and the types of products are increased continuously, and the water quality and the water quantity of discharged wastewater are increased year by year. Nitrogen is one of the main pollutants in the current petroleum refining and chemical wastewater, and ammonia nitrogen is the most common pollutant in the petrochemical wastewater. The excessive ammonia nitrogen in the water not only has toxic action on human and organisms, but also eutrophicates the water body, breeds bacteria and algae, and brings great difficulty to the treatment of domestic and industrial water, so that the removal of the ammonia nitrogen from petrochemical wastewater is necessary.
The microorganism treatment wastewater has the characteristics of strong comprehensive treatment capacity, simple and convenient treatment method, less secondary pollution and the like, but also has the problems of serious strain loss, small flora concentration and unstable reaction system which is easily influenced by the outside. The immobilized microorganism technology can solve the problem, the common microorganism immobilization technology has two categories of physical methods and chemical methods, but the cross-linking agent used by the traditional chemical method has the defects of certain toxicity, complex operation and the like, and the traditional physical method also has the problems of low treatment efficiency, long degradation time and the like, for example, in the CN201510557050.4 patent of 'an ammonia nitrogen degrading bacteria immobilization method for treating ammonia nitrogen wastewater', the degradation rate of immobilized high ammonia nitrogen degrading bacteria on ammonia nitrogen is 80-90%, the degradation time is 60h, and the degradation time is too long to be further improved.
Disclosure of Invention
The invention provides a preparation method of immobilized pellets of ammonia nitrogen degrading bacteria, aiming at the problems of high ammonia nitrogen concentration, serious thallus loss, long degradation time and the like in petrochemical wastewater treatment.
The technical scheme adopted by the invention is as follows: a preparation method of immobilized beads of ammonia nitrogen degrading bacteria comprises the following specific steps:
(1) activating the strain of the ochrobactrum anthropi to prepare a seed solution, and fermenting and culturing to obtain an ochrobactrum anthropi fermentation liquid;
(2) preparing a bacterial suspension: centrifuging the fermentation liquor obtained in the step (1) to collect bacterial sludge, washing with sterile normal saline, and preparing the cleaned bacterial sludge into the bacterial sludge with the effective viable count of 5 multiplied by 10 by using the sterile normal saline9～7×109CFU/mL of bacterial suspension;
(3) preparing immobilized ammonia nitrogen degrading bacteria: adding sodium alginate, sodium carboxymethylcellulose and ammonia nitrogen adsorbing material into deionized water, heating to dissolve, stirring uniformly, making into viscous wall material slurry, and cooling; adding the bacterial suspension obtained in the step (2) into the cooled thick wall material slurry, and uniformly stirring to prepare a mixed solution containing bacteria; then dripping the mixed solution containing the bacteria into AlCl3Standing the mixture in a cross-linking agent of the sterile aqueous solution at a low temperature for 6-12 hours to obtain immobilized beads; wherein the mass concentration of the sodium alginate in the viscous wall material slurry is 1.0-2.0%, the mass concentration of the sodium carboxymethyl cellulose is 0.5-1.5%, and the mass concentration of the ammonia nitrogen adsorbing material is 0.4-0.8%.
Preferably, the strain activation in the step (1) is as follows: inoculating ochrobactrum into an activation culture medium with the loading volume of 15-20%, controlling the temperature of activation culture to be 28-32 ℃, the pH to be 7.0-8.0 and the rotating speed to be 120-150 r/min, and performing activation culture for 36-48 h to obtain a seed solution, wherein the activation culture medium comprises the following components: 5.0-10.0 g/L of peptone, 1.0-5.0 g/L of yeast powder and 1.0-5.0 g/L of NaCl1.
Preferably, the fermentation culture in step (1) is: inoculating the seed liquid into a fermentation culture medium in an inoculation amount of 10-20% by volume ratio, and controlling fermentationFermenting and culturing at the temperature of 28-32 ℃, the pH value of 7.0-8.0 and the rotating speed of 120-150 r/min for 36-48 h to obtain fermentation liquor; wherein the fermentation medium comprises the following components: NH (NH)4Cl 0.765～1.146g/L、MgSO4 0.4～0.6g/L、K2HPO4 0.8～1.2g/L、KH2PO40.4-0.6 g/L, 8.0-10.0 g/L, FeSO g sodium succinate40.3～0.5g/L、NaCl0.3～0.5g/L。
Preferably, in the step (2), the centrifugal rotating speed is 6000-8000 r/min, and the centrifugal time is 3-5 min.
Preferably, the ammonia nitrogen adsorbing material in the step (3) is any one of artificial zeolite, kaolin and activated carbon; and (3) heating at the temperature of 60-80 ℃.
Preferably, the volume ratio of the ochrobactrum anthropi suspension to the viscous wall material slurry in the step (3) is 1: (3-5).
Preferably, the cross-linking agent AlCl in the step (3)3In a sterile aqueous solution of AlCl3The mass concentration of (A) is 1-3%; preferably, the low temperature in the step (3) is 0-4 ℃.
The strain is ochrobactrum anthropi purchased from Shanghai Bohu Biotechnology Limited. The ammonia nitrogen degrading bacteria immobilized pellet (w): petrochemical ammonia nitrogen wastewater (v) ═ 1 (5-10)) is weighed and placed in the petrochemical ammonia nitrogen wastewater, wherein the ammonia nitrogen concentration is 200-300 mg/L, and the ammonia nitrogen degradation rate is measured after shaking culture at 28-30 ℃ for 24-48 h.
Has the advantages that:
the immobilized ammonia nitrogen degrading bacteria provided by the invention has the synergistic effect of the components through the scientific proportion of sodium alginate, sodium carboxymethylcellulose, ammonia nitrogen adsorbing material and bacteria liquid. The sodium alginate, the sodium carboxymethylcellulose and the ammonia nitrogen adsorbing material form a space network, the structure is compact so as to fully protect thalli, a survival matrix is provided for microorganisms, the loss of thalli is reduced, and the obtained material has good stability and reusability and can efficiently treat ammonia nitrogen petrochemical wastewater.
Detailed Description
The present invention is further explained by the following examples, which are not intended to limit the present invention in any way.
Example 1:
(1) activation culture: inoculating ochrobactrum into an activation culture medium, wherein the filling volume is 15%, the activation culture temperature is 28 ℃, the pH is 7.0, and the rotating speed is 120 r/min; after activated culture for 36h, obtaining seed liquid; wherein the activation medium component (g/L): peptone 5.0, yeast powder 1.0 and NaCl1.0.
(2) Fermentation culture: inoculating the seed solution obtained in the step (1) into a fermentation culture medium with the inoculation amount of 10% (w/w), wherein the fermentation culture temperature is 28 ℃, the pH value is 7.0, and the rotation speed is 120 r/min; after fermentation culture for 36h, obtaining fermentation liquor; wherein the fermentation medium components (g/L): NH (NH)4Cl 0.765、MgSO4 0.4、K2HPO40.8、KH2PO40.4, sodium succinate 8.0, FeSO40.3、NaCl 0.3。
(3) Preparing a bacterial suspension: centrifuging the fermentation liquor obtained in the step (1) at 6000r/min for 3min, collecting bacterial sludge, washing with sterile normal saline, and repeating the steps for 3 times; then preparing the cleaned bacterial mud into 5 multiplied by 10 effective viable count by using sterile normal saline9CFU/mL of bacterial suspension.
(4) Preparing immobilized ammonia nitrogen degrading bacteria: adding sodium alginate, sodium carboxymethylcellulose and artificial zeolite into deionized water, heating at 60 deg.C for dissolving, and stirring to obtain viscous wall material slurry, wherein the wall material slurry contains sodium alginate 1.0 wt%, sodium carboxymethylcellulose 0.5 wt%, and artificial zeolite 0.4 wt%; adding the suspension of the ochrobactrum into the thick wall material slurry (bacterial suspension (v): thick wall material slurry (v): 1:3) which is cooled to room temperature, and uniformly stirring to prepare a mixed solution containing bacteria; ③ adding the mixed solution containing the bacteria into AlCl with the mass concentration of 1 percent dropwise3Standing the mixture for 6 hours at the temperature of 0 ℃ in the sterile water crosslinking agent to obtain the immobilized beads. Weighing immobilized pellets (w): petrochemical ammonia nitrogen wastewater (v): 1:5) and placing the pellets in the petrochemical ammonia nitrogen wastewater, wherein the ammonia nitrogen concentration is 200mg/L, performing shake culture at 28 ℃ for 24h, and the ammonia nitrogen degradation rate reaches 88.4%.
Example 2:
(1) activation culture: inoculating ochrobactrum into an activation culture medium, wherein the filling volume is 18%, the activation culture temperature is 30 ℃, the pH is 7.5, and the rotating speed is 135 r/min; after activation culture for 40h, obtaining seed liquid; wherein the activation medium component (g/L): peptone 8.0, yeast powder 3.0, NaCl 3.0.
(2) Fermentation culture: inoculating the seed solution obtained in the step (1) into a fermentation culture medium with an inoculation amount of 15% (w/w), wherein the fermentation culture temperature is 30 ℃, the pH value is 7.5, and the rotation speed is 135 r/min; fermenting and culturing for 40h to obtain fermentation liquor; wherein the fermentation medium components (g/L): NH (NH)4Cl 0.956、MgSO4 0.5、K2HPO41.0、KH2PO40.5, sodium succinate 9.0, FeSO40.4、NaCl 0.4。
(3) Preparing a bacterial suspension: centrifuging the fermentation liquor 7000r/min in the step (1) for 4min, collecting bacterial sludge, washing with sterile normal saline, and repeating the steps for 3 times; then preparing the cleaned bacterial mud into the effective viable count of 6 multiplied by 10 by using the sterile normal saline9CFU/mL of bacterial suspension.
(4) Preparing immobilized ammonia nitrogen degrading bacteria: adding sodium alginate, sodium carboxymethyl cellulose and kaolin into deionized water, heating at 70 ℃ to dissolve and uniformly stirring to prepare viscous wall material slurry, wherein the mass concentration of the sodium alginate in the wall material slurry is 1.5%, the mass concentration of the sodium carboxymethyl cellulose is 1.0%, and the mass concentration of the kaolin is 0.6%; adding the suspension of the ochrobactrum in the step (2) into the thick wall material slurry (bacterial suspension (v): thick wall material slurry (v): 1:4) which is cooled to room temperature, and uniformly stirring to prepare a mixed solution containing bacteria; ③ adding the mixed solution containing the bacteria into AlCl drop by drop3The immobilized beads are obtained by standing the mixture for 10 hours at the temperature of 2 ℃ in a cross-linking agent of sterile water with the mass concentration of 2%. Weighing immobilized pellets (w): petrochemical ammonia nitrogen wastewater (v): 1: 8), placing the immobilized pellets in the petrochemical ammonia nitrogen wastewater, wherein the ammonia nitrogen concentration is 250mg/L, performing shake culture at 29 ℃ for 36h, and the ammonia nitrogen degradation rate reaches 92.1%.
Example 3:
(1) activation culture: inoculating ochrobactrum into an activation culture medium, wherein the volume of a liquid filling volume is 20%, the activation culture temperature is 32 ℃, the pH value is 8.0, and the rotating speed is 150 r/min; activating and culturing for 48h to obtain seed liquid; wherein the activation medium component (g/L): peptone 10.0, yeast powder 5.0, NaCl 5.0.
(2) Fermentation culture: inoculating the seed solution obtained in the step (1) into a fermentation culture medium with an inoculation amount of 20% (w/w), wherein the fermentation culture temperature is 32 ℃, the pH value is 8.0, and the rotation speed is 150 r/min; fermenting and culturing for 48h to obtain fermentation liquor; wherein the fermentation medium components (g/L): NH (NH)4Cl 1.146、MgSO40.6、K2HPO41.2、KH2PO40.6, trisodium citrate 10.0, FeSO40.5、NaCl 0.5。
(3) Preparing a bacterial suspension: centrifuging the fermentation liquor 8000r/min obtained in the step (1) for 5min, collecting bacterial sludge, washing the bacterial sludge with sterile normal saline, and repeating the steps for 3 times; then using sterile physiological saline to prepare the cleaned bacterial mud into the bacterial mud with the effective viable count of 7 multiplied by 109CFU/mL of bacterial suspension.
(4) Preparing immobilized ammonia nitrogen degrading bacteria: adding sodium alginate, sodium carboxymethylcellulose and active carbon into deionized water, heating at 80 ℃ for dissolving, and uniformly stirring to prepare viscous wall material slurry, wherein the mass concentration of the sodium alginate is 2%, the mass concentration of the sodium carboxymethylcellulose is 1.5%, and the mass concentration of the active carbon is 0.8%; adding the suspension of the ochrobactrum in the step (2) into the cooled thick wall material slurry (the bacterial suspension (v): thick wall material slurry (v): 1:5), and uniformly stirring to prepare a mixed solution containing bacteria; ③ adding the mixed solution containing the bacteria into AlCl drop by drop3And standing the mixture for 12 hours at 4 ℃ in a crosslinking agent of sterile water with the mass concentration of 3% to obtain the immobilized beads. Weighing immobilized pellets (w): petrochemical ammonia nitrogen wastewater (v): 1: 10) and placing the pellets into the petrochemical ammonia nitrogen wastewater, wherein the ammonia nitrogen concentration is 300mg/L, and performing shake culture at 30 ℃ for 48h, wherein the ammonia nitrogen degradation rate reaches 90.5%.
Claims (7)
1. A preparation method of immobilized beads of ammonia nitrogen degrading bacteria comprises the following specific steps:
(1) activating the strain of the ochrobactrum anthropi to prepare a seed solution, and fermenting and culturing to obtain an ochrobactrum anthropi fermentation liquid;
(2) preparing a bacterial suspension: will be described in detail(1) The fermented liquid is centrifuged to collect bacterial mud, washed with sterile normal saline, and then the washed bacterial mud is prepared into the effective viable count of 5 multiplied by 10 by sterile normal saline9～7×109CFU/mL of bacterial suspension;
(3) preparing immobilized ammonia nitrogen degrading bacteria: adding sodium alginate, sodium carboxymethylcellulose and ammonia nitrogen adsorbing material into deionized water, heating to dissolve, stirring uniformly, making into viscous wall material slurry, and cooling; adding the bacterial suspension obtained in the step (2) into the cooled thick wall material slurry, and uniformly stirring to prepare a mixed solution containing bacteria; then dripping the mixed solution containing the bacteria into AlCl3Standing the mixture in a cross-linking agent of the sterile aqueous solution at a low temperature for 6-12 hours to obtain immobilized beads; wherein the mass concentration of the sodium alginate in the viscous wall material slurry is 1.0-2.0%, the mass concentration of the sodium carboxymethyl cellulose is 0.5-1.5%, and the mass concentration of the ammonia nitrogen adsorbing material is 0.4-0.8%.
2. The method of claim 1, wherein: the strain activation in the step (1) is as follows: inoculating ochrobactrum into an activation culture medium with the loading volume of 15-20%, controlling the temperature of activation culture to be 28-32 ℃, the pH to be 7.0-8.0 and the rotating speed to be 120-150 r/min, and performing activation culture for 36-48 h to obtain a seed solution, wherein the activation culture medium comprises the following components: 5.0-10.0 g/L of peptone, 1.0-5.0 g/L of yeast powder and 1.0-5.0 g/L of NaCl1.
3. The method of claim 1, wherein: the fermentation culture in the step (1) comprises the following steps: inoculating the seed solution into a fermentation culture medium by an inoculation amount of 10-20% by volume, controlling the fermentation culture temperature to be 28-32 ℃, the pH to be 7.0-8.0 and the rotating speed to be 120-150 r/min, and performing fermentation culture for 36-48 h to obtain a fermentation liquid; wherein the fermentation medium comprises the following components: NH (NH)4Cl 0.765～1.146g/L、MgSO4 0.4～0.6g/L、K2HPO4 0.8～1.2g/L、KH2PO40.4-0.6 g/L, 8.0-10.0 g/L, FeSO g sodium succinate40.3～0.5g/L、NaCl 0.3～0.5g/L。
4. The method of claim 1, wherein: in the step (2), the centrifugal rotating speed is 6000-8000 r/min, and the centrifugal time is 3-5 min.
5. The method of claim 1, wherein: the ammonia nitrogen adsorbing material in the step (3) is any one of artificial zeolite, kaolin or activated carbon; the heating temperature in the step (3) is 60-80 ℃.
6. The method of claim 1, wherein: the volume ratio of the bacterial suspension to the viscous wall material slurry in the step (3) is 1: (3-5).
7. The method of claim 1, wherein: AlCl in the cross-linking agent in the step (3)3The mass concentration of (A) is 1-3%; in the step (3), the low temperature is 0-4 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111319297.4A CN113980950A (en) | 2021-11-09 | 2021-11-09 | Preparation method of ammonia nitrogen degrading bacteria immobilized pellet |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111319297.4A CN113980950A (en) | 2021-11-09 | 2021-11-09 | Preparation method of ammonia nitrogen degrading bacteria immobilized pellet |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113980950A true CN113980950A (en) | 2022-01-28 |
Family
ID=79747251
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111319297.4A Pending CN113980950A (en) | 2021-11-09 | 2021-11-09 | Preparation method of ammonia nitrogen degrading bacteria immobilized pellet |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113980950A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928081A (en) * | 2006-09-28 | 2007-03-14 | 武汉益生泉生物科技开发有限责任公司 | Artificial biological film and preparation method |
| CN104593349A (en) * | 2015-01-14 | 2015-05-06 | 华南理工大学 | GY2B degrading bacteria immobilizing ball as well as preparation method and application thereof |
| CN105063009A (en) * | 2015-09-02 | 2015-11-18 | 博天环境集团股份有限公司 | Ammonia-nitrogen degrading bacterium immobilization method for treating ammonia-nitrogen wastewater |
-
2021
- 2021-11-09 CN CN202111319297.4A patent/CN113980950A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928081A (en) * | 2006-09-28 | 2007-03-14 | 武汉益生泉生物科技开发有限责任公司 | Artificial biological film and preparation method |
| US20080160596A1 (en) * | 2006-09-28 | 2008-07-03 | Qinsheng Li | Artificial biomembrane and the method for manufacturing the same |
| CN104593349A (en) * | 2015-01-14 | 2015-05-06 | 华南理工大学 | GY2B degrading bacteria immobilizing ball as well as preparation method and application thereof |
| CN105063009A (en) * | 2015-09-02 | 2015-11-18 | 博天环境集团股份有限公司 | Ammonia-nitrogen degrading bacterium immobilization method for treating ammonia-nitrogen wastewater |
Non-Patent Citations (3)
| Title |
|---|
| 孙霞 等: "聚乙烯醇-海藻酸钠-改性沸石固定化菌球降解氨氮的研究", 《化学与生物工程》 * |
| 王启山: "对硝基苯酚降解菌的固定化及其降解特性研究", 《中国优秀硕士论文库工程科技Ⅰ辑》 * |
| 陈坦林;赵薇;朱雪琴;朱鹏程;熊婷婷;: "CMC-硅藻土复合固定硝化细菌降解养殖水体中的氨氮", 江苏农业科学 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108359663B (en) | Phosphorus-accumulating bacterium immobilized pellet and application thereof | |
| Spettoli et al. | Immobilization of Leuconostoc oenos ML 34 in calcium alginate gels and its application to wine technology | |
| CN108624582B (en) | Microbial preparation for soil remediation | |
| CN112940732B (en) | Soil organic phosphorus pesticide degradation catalyst and preparation method thereof | |
| CN110643593A (en) | Preparation method of modified corn straw material immobilized agricultural microbial agent | |
| CN114014701B (en) | Method for strengthening aerobic fermentation of kitchen waste by using oil-removing microbial agent | |
| CN113462396A (en) | Biochar soil conditioner and preparation method and application thereof | |
| CN112011476B (en) | Preparation method of high-strength immobilized microspheres for embedding thiobacillus denitrificans | |
| CN111944799B (en) | Preparation method and application of immobilized particles embedded with Thiobacillus denitrificans | |
| CN111440786A (en) | Method for removing soil 2,4, 6-trichlorophenol by biomass charcoal immobilized high-efficiency degrading strain | |
| CN111072154A (en) | Preparation method of sewage treatment agent by using microorganisms | |
| CN112960766B (en) | Aerobic biomembrane material, preparation method and application thereof in sewage treatment technology | |
| CN109797120B (en) | Preparation method and application of microecological preparation for removing nitrate in soil | |
| CN113980950A (en) | Preparation method of ammonia nitrogen degrading bacteria immobilized pellet | |
| CN101165170A (en) | Atrazine herbicide degradation bacterium and preparation method for bacterium preparation of the same | |
| CN111115849A (en) | Preparation method of microbial sewage purification treatment agent | |
| CN105586331A (en) | Method of denitrified composite microbial inoculant | |
| CN115960783A (en) | Anaerobic microorganism combination with sulfonamide antibiotic degradation function and application thereof | |
| CN112921026B (en) | Granular composite microbial agent and preparation method thereof | |
| CN109231463A (en) | A kind of preparation and application of compound diatomite sewage-treating agent | |
| CN101392245B (en) | Preparation and application of flocculating alcaligenes faecalis fixed pellet | |
| CN114456811A (en) | Farmland carbon-based soil remediation agent and preparation method thereof | |
| CN110921841B (en) | Biochemical preparation for repairing starch industrial wastewater | |
| CN114031189A (en) | Method for treating sewage with low carbon-nitrogen ratio | |
| CN112694997A (en) | Compound microorganism bacterium stock solution suitable for sewage treatment and active solution culture method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20220128 |