CN102604865B - Harmless treatment method of coffeine-containing waste water and bacterium used in harmless treatment method - Google Patents
Harmless treatment method of coffeine-containing waste water and bacterium used in harmless treatment method Download PDFInfo
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- CN102604865B CN102604865B CN2012100675775A CN201210067577A CN102604865B CN 102604865 B CN102604865 B CN 102604865B CN 2012100675775 A CN2012100675775 A CN 2012100675775A CN 201210067577 A CN201210067577 A CN 201210067577A CN 102604865 B CN102604865 B CN 102604865B
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- xanthine
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- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 title claims abstract description 119
- 239000002351 wastewater Substances 0.000 title claims abstract description 98
- 238000000034 method Methods 0.000 title claims abstract description 43
- 241000894006 Bacteria Species 0.000 title description 22
- 241000589776 Pseudomonas putida Species 0.000 claims abstract description 40
- 238000004065 wastewater treatment Methods 0.000 claims abstract description 9
- LCCDINSFSOALJK-UHFFFAOYSA-N 1,3,4-trimethylpurine-2,6-dione Chemical compound O=C1N(C)C(=O)N(C)C2(C)N=CN=C21 LCCDINSFSOALJK-UHFFFAOYSA-N 0.000 claims description 90
- 238000011081 inoculation Methods 0.000 claims description 2
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 abstract description 14
- 229960001948 caffeine Drugs 0.000 abstract description 14
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 abstract description 14
- 238000004321 preservation Methods 0.000 abstract description 11
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- 238000012360 testing method Methods 0.000 description 9
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 244000005700 microbiome Species 0.000 description 7
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- 229910052760 oxygen Inorganic materials 0.000 description 7
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 6
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 6
- 239000004327 boric acid Substances 0.000 description 6
- 239000001110 calcium chloride Substances 0.000 description 6
- 229910001628 calcium chloride Inorganic materials 0.000 description 6
- 229910000365 copper sulfate Inorganic materials 0.000 description 6
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 6
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 6
- 229910000397 disodium phosphate Inorganic materials 0.000 description 6
- 235000019800 disodium phosphate Nutrition 0.000 description 6
- 230000004151 fermentation Effects 0.000 description 6
- 239000011790 ferrous sulphate Substances 0.000 description 6
- 235000003891 ferrous sulphate Nutrition 0.000 description 6
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 6
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 6
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 6
- 229910000368 zinc sulfate Inorganic materials 0.000 description 6
- 229960001763 zinc sulfate Drugs 0.000 description 6
- 239000000284 extract Substances 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 238000010564 aerobic fermentation Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
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- 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 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241001122767 Theaceae Species 0.000 description 2
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- 150000001413 amino acids Chemical class 0.000 description 2
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- 238000007254 oxidation reaction Methods 0.000 description 2
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- 239000003643 water by type Substances 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
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Images
Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
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- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses pseudomonas putida CT2025 with preservation department of CGMCC (China General Microbiological Culture Collection Center) and preservation address of No.3, No.1 Institution, Beichen West Road, Chaoyang District, Beijing, Chinese Academy of Sciences Institute of Microbiology, and preservation date of February 17th, 2012 and preservation number of CGMCC No.5767. The invention also discloses a harmless treatment method of coffeine-containing waste water. The method comprises the following steps of: inoculating 0.8-1.2kg of pseudomonas putida in every 100ton of caffeine-containing waste water, and intervening and applying in any one of the following waste water treatment stages: aerobic treatment stage after anaerobic fermentation and application after the aerobic treatment on the waste water is finished.
Description
Technical field
The invention belongs to biological technical field.Specifically, the present invention relates to a kind of method for innocent treatment that utilizes the degradation by bacteria trimethyl-xanthine to contain caffeine waste water.
Background technology
Tea-polyphenol is the important physiologically active ingredient that extracts from tealeaves, have anti-oxidant, radioprotective, anti-mutation, antitumor, the anti-ageing various functions of waiting for a long time, have a good application prospect at functional food and field of medicaments, therefore, tea-polyphenol extract and the purifying industry development very swift and violent, by 2011, national tea-polyphenol manufacturing enterprise was above 60, scale has more than 10 in the enterprise more than 100 tons, and annual production is more than 4000 tons.Tea-polyphenol often adopts the techniques such as extraction, purification, purification and drying to be prepared from, and wherein purification process is to affect the most important operation of quality product.The purification process main purpose of tea-polyphenol is the impurity such as removal trimethyl-xanthine.The main method that removes the impurity such as caffeine from tea-polyphenol has: organic solvent (such as chloroform etc.) extraction, precipitation by metallic ion are (such as Ca
2+Deng)/acid turns molten/ethyl acetate extraction, adverse current chromatogram such as separates at the several method with resin absorption, the each have their own relative merits of these methods, but all methods all can produce the tea-polyphenol that contains of some amount, trimethyl-xanthine, pectin, albumen, the waste water of amino acid and some colloidal particle things, tea-polyphenol wherein, pectin, albumen, amino acid and particulate matter are to the chemical oxygen demand (COD) of waste water, biological oxygen demand (BOD) (BOD), suspended substance (SS) and colourity etc. have material impact, are the major objective things in tea-polyphenol production and the refining enterprise wastewater innocent treatment procedure.At present, fairly large tea-polyphenol production and refining enterprise quite pay attention to wastewater treatment, the method of taking is generally: the wastewater collection in the production process is entered water collecting basin, add coagulating agent or flocculation agent to remove tea-polyphenol and colloidal suspension particle, carry out successively afterwards microorganism anaerobic fermentation and aerobic fermentation to remove the most of organism in the waste water, remove microorganism and release by the biological filter at last.
Trimethyl-xanthine has another name called caffeine, and white needle-like crystals has stronger central nervous excitation effect and diuretic properties, is widely used in the fields such as medicine, food, makeup.In tealeaves and sheet end byproduct thereof, caffeine content is generally 1-5%.In the waste water that extracts the generation of tea-polyphenol or tea-polyphenol treating process, all contain a considerable amount of trimethyl-xanthines, because the technique that adopts is different, content of caffeine does not wait to thousands of milligrams at every liter of hundreds of milligram in the waste water.Trimethyl-xanthine belongs to the control product in China, and its production must obtain strictly examining of relevant department.Common tea-polyphenol extracts or makes with extra care enterprise and generally is difficult to obtain to produce authorize.Therefore, tea-polyphenol manufacturing enterprise generally can not take recovery measure to caffeine contained in the production process waste water.Studies show that, even the trimethyl-xanthine of low concentration namely has obvious toxicity to aquatic animals and plants, so trimethyl-xanthine has very large Latent destruction effect to aquatic environment.Because trimethyl-xanthine all shows very strong resistance to soda acid and common oxidizer treatment, common microorganism generally also is difficult to degraded and utilizes caffeine, and be dissolved in caffeine good dispersity in the water, can not be as other pollutent water body chroma etc. be produced visual impact, therefore, generally also be difficult in the monitoring index of biological oxygen demand (BOD), suspended substance and colourity etc., be reflected even contain the caffeine of higher concentration in the waste water, thereby it is out in the cold to cause trimethyl-xanthine remaining in the waste water to be easy to.And, the present treatment technology that does not also effectively contain caffeine waste water.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of simple to operate, mild condition, the method for innocent treatment that contains trimethyl-xanthine waste water of effect stability and used pseudomonas putida, can utilize the biological fermentation approach efficiently to remove trimethyl-xanthine in the waste water according to method of the present invention.
In order to solve the problems of the technologies described above, the invention provides a kind of pseudomonas putida, the preservation name is called: pseudomonas putida (Pseudomonas putida) CT2025, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica; Preservation date: on 02 17th, 2012, preserving number: CGMCC NO.5767.
The present invention also provides the numerous method of expansion of above-mentioned pseudomonas putida simultaneously: pseudomonas putida (Pseudomonas putida) CGMCC NO.5767 is inoculated in expands in numerous liquid nutrient medium, under 28～30 ℃, 130～150 rev/mins conditions, expand numerous cultivation, when gained expands the absorbancy in 600 nanometers of numerous product 〉=4, stop to expand numerous cultivation.
Improvement as the numerous method of expansion of pseudomonas putida of the present invention:
Expanding numerous liquid nutrient medium is: every liter is expanded Repone K, the ferrous sulfate of 1.5 mmoles, the sal epsom of 1.5 mmoles, the calcium chloride of 0.4 mmole, the Sodium phosphate dibasic of 0.3 mmole, 0.15 micromolar boric acid, 0.05 micromolar copper sulfate, 0.05 micromolar manganous sulfate, 0.05 micromolar Sodium orthomolybdate, 0.05 micromolar zinc sulfate, sucrose 10 grams, peptone 10 grams, trimethyl-xanthine 2.5～7.5 mmoles that contain 5.0 mmoles in numerous liquid nutrient medium, and all the other are water; After regulating pH7.0, high-temperature sterilization.
The present invention also provides a kind of method for innocent treatment that contains trimethyl-xanthine waste water simultaneously: per 100 tons pseudomonas putida (fresh weight) that inoculates 0.8～1.2kg in the trimethyl-xanthine waste water that contains, get involved application in the stage of following any one wastewater treatment:
The aerobic treatment stage after anaerobically fermenting is used,
After finishing, the waste water aerobic treatment uses;
Above-mentioned pseudomonas putida is pseudomonas putida (Pseudomonas putida) CGMCC NO.5767.
The numerous product of expansion of the absorbancy in 600 nanometers of gained of the present invention 〉=4 (namely comprising the pseudomonas putida (Pseudomonas putida) of expanding numerous liquid nutrient medium) both can be used for bacterial classification to be preserved, and also is used for containing the processing of trimethyl-xanthine waste water.The method that bacterial classification is preserved is, adds the dimethyl sulfoxide (DMSO) that is equivalent to bacteria liquid long-pending 20% in the bacterium liquid numerous product of expansion of the absorbancy of 600 nanometers 〉=4 (namely), places minus 20 degrees～subzero 70 to spend Refrigerator stores.
Contain in the method for innocent treatment of trimethyl-xanthine waste water in the present invention, pseudomonas putida (fresh weight) refers to: with the above-mentioned bacterium liquid numerous product of expansion of the absorbancy of 600 nanometers 〉=4 (namely) centrifugal 10-15 minute collected bacterial precipitation under 2000-5000 rev/min condition.
The present invention is applicable to Wastewater Treated by Activated Sludge Process and contains trimethyl-xanthine waste water, can get involved at the three phases of wastewater treatment and use: both can after coagulation or flocculation, use in the waste water, also can use in the aerobic treatment stage after anaerobically fermenting, also can after conventional waste water aerobic treatment finishes, use (seeing Fig. 1).Optimize as the intervention stage: the aerobic treatment stage of this bacterium behind anaerobically fermenting gets involved, not only the trimethyl-xanthine degraded thoroughly, and can with original waste water treatment process seamless connection, substantially without additional cost.Getting involved the method for using is: when using first (when dropping into bacterium first), can be according to the size of aerobic fermentation aeration tank, ratio according to 100 tons of waste water of 1 kilogram of bacterium (fresh weight) is inoculated in the conventional aerobic aeration tank, conventional aeration adapts to 3-5 days (namely after the pseudomonas putida access, waste water is carried out conventional aeration oxygen replenishing fermentation 3-5 days, normal temperature (being generally 15-30 ℃) gets final product), be colonizated in the mud in order to this bacterium.When again processing waste water afterwards, can contain according to the sewage aerobic treatment method of routine the harmless treatment of trimethyl-xanthine waste water, that is, the treatment time of waste water in aerobic aeration tank (tank) is 48～160 hours, and treatment temp is controlled at 15-30 ℃.When the trimethyl-xanthine concentration in the waste water was lower than 500 mg/litre, the treatment time in aerobic aeration tank (tank) was 48 hours; When the trimethyl-xanthine concentration in the waste water was the 500-1500 mg/litre, the treatment time in aerobic aeration tank (tank) was 96 hours; When the trimethyl-xanthine concentration in the waste water surpassed 1500 mg/litre, the treatment time in aerobic aeration tank (tank) was 96～160 hours.Therefore, when trimethyl-xanthine concentration was lower than 500 mg/litre in the waste water, can thoroughly degrade every day with being equivalent to access in the waste water of 50,000 times of bacterial classification amounts trimethyl-xanthine; When trimethyl-xanthine concentration in the waste water during in the 500-1500 mg/litre, can thoroughly degrade every day with being equivalent to access in 2.5 ten thousand times the waste water of bacterial classification amount trimethyl-xanthine; When trimethyl-xanthine concentration in the waste water surpasses 1500 mg/litre, the trimethyl-xanthine that is equivalent to get involved in ten thousand times of waste water of 1.5-2.5 of bacterial classification amount thoroughly can be degraded.Optimum temperuture should be controlled at 15-30 ℃ during wastewater treatment.
Principal feature of the present invention is: 1) utilize the characteristics that can efficiently decompose trimethyl-xanthine from the pseudomonas putida of tea garden soil separation, carried out biological degradation and harmless treatment that general chemistry oxidation and microbial fermentation are difficult to the trimethyl-xanthine of removal; 2) produce the refined wastewater treatment process with conventional tea-polyphenol and be connected well, almost drop into without extra; 3) can get involved in the aerobic fermentation stage in original waste water treatment process, trimethyl-xanthine is removed thoroughly.4), method of the present invention be applicable to remove tea-polyphenol extracts or the tea-polyphenol treating process in produce contain caffeine in the caffeine waste water.
Description of drawings
Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail.
Fig. 1 is that the waste water conventional activated sludge method that tea-polyphenol production of the present invention and treating process produce is processed and trimethyl-xanthine decomposer (being pseudomonas putida) intervention stage schematic diagram;
Among the figure: the necessary operation of solid arrow representative, dotted arrow represents optional operation; Trilateral represents three kinds of optional trimethyl-xanthine decomposers intervention stages, as preferably, gets involved in the aerobic fermentation stage, and not only the trimethyl-xanthine degraded is thorough, with the original process seamless connection, and almost without extra input.
Embodiment
Purifying and the domestication of embodiment 1, pseudomonas putida (being the caffeine decomposer):
Gather the root topsoil from tea place (being arranged in Tea Germplasm garden, the Zhejiang University Hua Jiachi school district) soil of planting more than 10 years, Litter removal, 1 gram soil is added 1 liter of stirring of sterilized water, get 0.1 milliliter of supernatant, be inoculated in and be equipped with 20 milliliters take trimethyl-xanthine as sole carbon source and the solid bacteria isolation medium of nitrogenous source (5.0 mM/ls of Repone K, 1.5 mM/l ferrous sulfate, 1.5 mM/l sal epsom, 0.4 mM/l calcium chloride, 0.3 mM/l Sodium phosphate dibasic, 0.15 micromoles per liter boric acid, 0.05 micromoles per liter copper sulfate, 0.05 micromoles per liter manganous sulfate, 0.05 micromoles per liter Sodium orthomolybdate, 0.05 micromoles per liter zinc sulfate, 25 mM/ls of trimethyl-xanthines, agar 12 grams per liters are in culture dish pH7.0); Under 28-30 ℃ of condition, be inverted and cultivated 3 days, each mono-clonal bacterium bacterial plaque of picking white, respectively inoculation enter 1 milliliter take trimethyl-xanthine as sole carbon source and the liquid purifying substratum of nitrogenous source (100 mM/ls of trimethyl-xanthines, do not add agar, all the other compositions and acidity are with the solid bacteria isolation medium), shaking culture is 3 days under 28-30 ℃, 130-150 rev/min condition, screening can be on this liquid purifying substratum the bacterial strain of normal growth.The bacterial strain that obtains is inoculated according to the ratio of 1 liter of liquid purifying substratum of 2 milliliters of bacterium liquid, and succeeding transfer culture is 3 days under above-mentioned the same terms; Subculture 10-15 time (for example being 15 times) obtains the trimethyl-xanthine decomposer (that is, pseudomonas putida) of taming so continuously.
The preparation method of above-mentioned solid bacteria isolation medium is as follows: with the Repone K of 5.0 mmoles, 1.5 the ferrous sulfate of mmole, 1.5 the sal epsom of mmole, 0.4 the calcium chloride of mmole, 0.3 the Sodium phosphate dibasic of mmole, 0.15 micromolar boric acid, 0.05 micromolar copper sulfate, 0.05 micromolar manganous sulfate, 0.05 micromolar Sodium orthomolybdate, 0.05 micromolar zinc sulfate, the trimethyl-xanthine of 25 mmoles and 12 gram agar waters are settled to 1 liter, then be the sodium hydroxide adjusting pH to 7.0 of mass concentration 4% with concentration, in 121 ℃ of sterilization 20min, get the solid bacteria isolation medium.
The preparation method of aforesaid liquid pure medium is as follows: the trimethyl-xanthine water of the Sodium phosphate dibasic of the calcium chloride of the sal epsom of the ferrous sulfate of the Repone K of 5.0 mmoles, 1.5 mmoles, 1.5 mmoles, 0.4 mmole, 0.3 mmole, 0.15 micromolar boric acid, 0.05 micromolar copper sulfate, 0.05 micromolar manganous sulfate, 0.05 micromolar Sodium orthomolybdate, 0.05 micromolar zinc sulfate, 100 mmoles is settled to 1 liter, then be that 4% sodium hydroxide is regulated pH to 7.0 with mass concentration, in 121 ℃ of sterilization 20min, get the liquid purifying substratum.
The trimethyl-xanthine decomposer that obtains domestication is carried out preservation, the preservation name is called: pseudomonas putida (Pseudomonas putida) CT2025, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica; Preservation date: on 02 17th, 2012, preserving number: CGMCC NO.5767.
Embodiment 2, trimethyl-xanthine decomposer are expanded numerous:
Get the numerous liquid nutrient medium of the expansion that contains trimethyl-xanthine (5.0 mM/ls of Repone K that 1 liter of pseudomonas putida (Pseudomonas putida) CGMCC NO.5767 is inoculated in 250 liters, 1.5 mM/l ferrous sulfate, 1.5 mM/l sal epsom, 0.4 mM/l calcium chloride, 0.3 mM/l Sodium phosphate dibasic, 0.15 micromoles per liter boric acid, 0.05 micromoles per liter copper sulfate, 0.05 micromoles per liter manganous sulfate, 0.05 micromoles per liter Sodium orthomolybdate, 0.05 micromoles per liter zinc sulfate, sucrose 10 grams per liters, peptone 10 grams per liters, 7.5 mM/ls of trimethyl-xanthines, pH7.0), in 28-30 ℃, cultivate expansion under the 130-150 rev/min of condition numerous, when substratum during in the absorbancy of 600 nanometers 〉=4 (expand numerous time be about 2-4 days), can be used for containing the processing of trimethyl-xanthine waste water.
The preparation method of the numerous liquid nutrient medium of above-mentioned expansion is: with the Repone K of 5.0 mmoles, 1.5 the ferrous sulfate of mmole, 1.5 the sal epsom of mmole, 0.4 the calcium chloride of mmole, 0.3 the Sodium phosphate dibasic of mmole, 0.15 micromolar boric acid, 0.05 micromolar copper sulfate, 0.05 micromolar manganous sulfate, 0.05 micromolar Sodium orthomolybdate, 0.05 micromolar zinc sulfate, sucrose 10 grams, peptone 10 grams, trimethyl-xanthine 7.5 mmole waters are settled to 1 liter, then be that 4% sodium hydroxide is regulated pH to 7.0 with mass concentration, in 121 ℃ of sterilization 20min, must expand numerous liquid nutrient medium.
Embodiment 3, a kind of method for innocent treatment (being that trimethyl-xanthine decomposer (pseudomonas putida) processing contains trimethyl-xanthine waste water) that contains trimethyl-xanthine waste water:
The waste water conventional activated sludge method that conventional tea-polyphenol production and treating process produce is processed according to " heterogeneous catalytic oxidation and cyclic activated sludge system (CASS) combination process is processed the refining factory effluent of tea-polyphenol " (Zhang Bo, Xiao Qianfen, the summer cleaning politics, economics, organization, and ideology. water purification technology, 2011,30 (2): 33-36) carry out.
With expanding Bacteria liquid (embodiment 2 gains) after numerous under 2000-2500 rev/min of condition centrifugal 15 minutes, collect bacterial precipitation, as pseudomonas putida (fresh weight).It is in 10 tons the aerobic aeration tank (tank) that 0.1 kilogram of pseudomonas putida (fresh weight) is inoculated in volume, after passing into the waste water behind the anaerobically fermenting, carry out conventional aeration and adapt to field planting 3 days, namely after the pseudomonas putida access, waste water is carried out conventional aeration oxygen replenishing fermentation 3 days, normal temperature gets final product, and is colonizated in the mud in order to this bacterium.When again processing waste water afterwards, can contain according to the sewage aerobic treatment method of routine the harmless treatment of trimethyl-xanthine waste water, that is, the treatment time of waste water in aerobic aeration tank (tank) is 48～160 hours, and treatment temp is controlled at 15-30 ℃.Specific as follows:
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance is lower than 500 mg/litre, treatment time in aerobic aeration tank (tank) is 48 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, can thoroughly degrade trimethyl-xanthine in 5 tons of waste water every day;
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance is the 500-1500 mg/litre, treatment time in aerobic aeration tank (tank) is 96 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, trimethyl-xanthine in 2.5 tons of waste water thoroughly can be degraded every day;
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance surpasses 1500 mg/litre, treatment time in aerobic aeration tank (tank) is 96～160 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, trimethyl-xanthine in the 1.5-2.5 ton waste water thoroughly can be degraded every day.
Embodiment 4, a kind of method for innocent treatment (being that the processing of trimethyl-xanthine decomposer contains trimethyl-xanthine waste water) that contains trimethyl-xanthine waste water:
With expanding Bacteria liquid (embodiment 2 gains) after numerous under 3000-3500 rev/min of condition centrifugal 12 minutes, collect bacterial precipitation, as pseudomonas putida (fresh weight).It is in 50 tons the aerobic aeration tank (tank) that 0.5 kilogram of pseudomonas putida (fresh weight) is inoculated in volume, after passing into the waste water behind the anaerobically fermenting, carry out conventional aeration and adapt to field planting 4 days, namely after the pseudomonas putida access, waste water is carried out conventional aeration oxygen replenishing fermentation 4 days, normal temperature gets final product, and is colonizated in the mud in order to this bacterium.When again processing waste water afterwards, can contain according to the sewage aerobic treatment method of routine the harmless treatment of trimethyl-xanthine waste water, that is, the treatment time of waste water in aerobic aeration tank (tank) is 48～160 hours, and treatment temp is controlled at 15-30 ℃.Specific as follows:
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance is lower than 500 mg/litre, treatment time in aerobic aeration tank (tank) is 48 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, can thoroughly degrade trimethyl-xanthine in 25 tons of waste water every day;
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance is the 500-1500 mg/litre, treatment time in aerobic aeration tank (tank) is 96 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, trimethyl-xanthine in 12.5 tons of waste water thoroughly can be degraded every day;
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance surpasses 1500 mg/litre, treatment time in aerobic aeration tank (tank) is 96～160 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, trimethyl-xanthine in the 7.5-12.5 ton waste water thoroughly can be degraded every day.
Embodiment 5, a kind of method for innocent treatment (being that the processing of trimethyl-xanthine decomposer contains trimethyl-xanthine waste water) that contains trimethyl-xanthine waste water:
With expanding Bacteria liquid (embodiment 2 gains) after numerous under 4500-5000 rev/min of condition centrifugal 10 minutes, collect bacterial precipitation, as pseudomonas putida (fresh weight).It is in 100 tons the aerobic aeration tank (tank) that 1 kilogram of pseudomonas putida (fresh weight) is inoculated in volume, after passing into the waste water behind the anaerobically fermenting, carry out conventional aeration and adapt to field planting 5 days, namely after the pseudomonas putida access, waste water is carried out conventional aeration oxygen replenishing fermentation 5 days, normal temperature gets final product, and is colonizated in the mud in order to this bacterium.When again processing waste water afterwards, can contain according to the sewage aerobic treatment method of routine the harmless treatment of trimethyl-xanthine waste water, that is, the treatment time of waste water in aerobic aeration tank (tank) is 48～160 hours, and treatment temp is controlled at 15-30 ℃.Specific as follows:
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance is lower than 500 mg/litre, treatment time in aerobic aeration tank (tank) is 48 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, can thoroughly degrade trimethyl-xanthine in 50 tons of waste water every day;
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance is the 500-1500 mg/litre, treatment time in aerobic aeration tank (tank) is 96 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, trimethyl-xanthine in 25 tons of waste water thoroughly can be degraded every day;
When the trimethyl-xanthine concentration in the waste water after detecting anaerobically fermenting in advance surpasses 1500 mg/litre, treatment time in aerobic aeration tank (tank) is 96～160 hours, waste water after the processing is trimethyl-xanthine concentration≤1 mg/litre after testing, that is, trimethyl-xanthine in the 15-25 ton waste water thoroughly can be degraded every day.
Annotate: the detection method of trimethyl-xanthine concentration is carried out according to GB/T 8312--2002 in the above-mentioned waste water.
At last, it is also to be noted that, what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (1)
1. method for innocent treatment that contains trimethyl-xanthine waste water is characterized in that: contain the pseudomonas putida that inoculation in the trimethyl-xanthine waste water enters 0.8 ~ 1.2kg per 100 tons, get involved in the stage of following any one wastewater treatment and use:
The aerobic treatment stage after anaerobically fermenting is used,
After finishing, the waste water aerobic treatment uses;
Above-mentioned pseudomonas putida is pseudomonas putida (Pseudomonas putida) CGMCC NO.5767.
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