CN102604865A - 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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- CN102604865A CN102604865A CN2012100675775A CN201210067577A CN102604865A CN 102604865 A CN102604865 A CN 102604865A CN 2012100675775 A CN2012100675775 A CN 2012100675775A CN 201210067577 A CN201210067577 A CN 201210067577A CN 102604865 A CN102604865 A CN 102604865A
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- waste water
- pseudomonas putida
- trimethyl
- 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 112
- 239000002351 wastewater Substances 0.000 title claims abstract description 98
- 238000000034 method Methods 0.000 title claims abstract description 44
- 241000894006 Bacteria Species 0.000 title description 22
- 241000589776 Pseudomonas putida Species 0.000 claims abstract description 46
- 238000004321 preservation Methods 0.000 claims abstract description 13
- 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 91
- 239000007788 liquid Substances 0.000 claims description 22
- 235000015097 nutrients Nutrition 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 7
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 7
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 7
- 239000004327 boric acid Substances 0.000 claims description 7
- 239000001110 calcium chloride Substances 0.000 claims description 7
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 7
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 7
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 7
- 229940061607 dibasic sodium phosphate Drugs 0.000 claims description 7
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 7
- 239000011790 ferrous sulphate Substances 0.000 claims description 7
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 7
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 7
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 7
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 7
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims description 7
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 7
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 7
- 229960001763 zinc sulfate Drugs 0.000 claims description 7
- 244000005700 microbiome Species 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 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
- 229930006000 Sucrose Natural products 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 238000011081 inoculation Methods 0.000 claims description 2
- 101000794816 Pseudomonas putida Anthranilate synthase component 1 Proteins 0.000 claims 1
- 101000847784 Pseudomonas putida Anthranilate synthase component 2 Proteins 0.000 claims 1
- 238000000855 fermentation Methods 0.000 abstract description 8
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 abstract description 3
- 229960001948 caffeine Drugs 0.000 abstract description 3
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 abstract description 3
- 238000009629 microbiological culture Methods 0.000 abstract 1
- 238000005273 aeration Methods 0.000 description 29
- 238000012545 processing Methods 0.000 description 14
- 230000001580 bacterial effect Effects 0.000 description 12
- 230000003203 everyday effect Effects 0.000 description 11
- 230000008569 process Effects 0.000 description 11
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 7
- 230000004151 fermentation Effects 0.000 description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 239000001301 oxygen 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
- 238000010564 aerobic fermentation Methods 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 239000012467 final product Substances 0.000 description 4
- 238000002955 isolation Methods 0.000 description 4
- 238000007670 refining Methods 0.000 description 4
- 239000010865 sewage Substances 0.000 description 4
- 239000010802 sludge Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 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
- 244000269722 Thea sinensis Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005189 flocculation Methods 0.000 description 2
- 230000016615 flocculation Effects 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000001814 pectin Substances 0.000 description 2
- 229920001277 pectin Polymers 0.000 description 2
- 235000010987 pectin Nutrition 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 235000009024 Ceanothus sanguineus Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 240000003553 Leptospermum scoparium Species 0.000 description 1
- 235000015459 Lycium barbarum Nutrition 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
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000000701 coagulant Substances 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000001882 diuretic effect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000003810 ethyl acetate extraction Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 150000001455 metallic ions Chemical class 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 239000013618 particulate matter Substances 0.000 description 1
- 230000004223 radioprotective effect Effects 0.000 description 1
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- 238000011160 research Methods 0.000 description 1
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- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
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
Landscapes
- 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 date of February 17th, 2012 and preservation number of CGMCC (China General Microbiological Culture Collection Center) 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 theine waste water.
Background technology
Tea-polyphenol is the important physiological activeconstituents that from tealeaves, extracts, and has anti-oxidant, radioprotective, mutation, antitumor, anti-ageing many functions of waiting for a long time, and has a good application prospect at functional food and field of medicaments; Therefore; Tea-polyphenol extracts with the purifying industry development very swift and violent, and by 2011, national tea-polyphenol manufacturing enterprise was above 60 families; Scale the enterprise more than 100 tons have 10 surplus family, YO is more than 4000 tons.Tea-polyphenol often adopts prepared such as extraction, purification, purification and drying to form, and wherein purification process is to influence the most important operation of quality product.The purification process main purpose of tea-polyphenol is impurity such as removal trimethyl-xanthine.The main method that from tea-polyphenol, removes impurity such as theine has: organic solvent (like chloroform etc.) extraction, precipitation by metallic ion are (like Ca
2+Deng)/acid change to dissolve/and ethyl acetate extraction, adverse current chromatogram several method such as separate with resin absorption; The each have their own relative merits of these methods; But all methods all can produce the waste water that contains tea-polyphenol, trimethyl-xanthine, pectin, albumen, amino acid and some colloidal particle things of some amount; Wherein tea-polyphenol, pectin, albumen, amino acid and PM for particulate matter have material impact to chemical oxygen demand (COD), BOD (BOD), suspended substance (SS) and the colourity etc. of waste water, 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: water collecting basin is gone in the wastewater collection in the production process; Add coagulating agent or flocculation agent with removal tea-polyphenol and colloidal suspension particle, carry out mikrobe anaerobic fermentation and aerobic fermentation afterwards successively, remove mikrobe and release through the biological filter at last to remove the most of organism in the waste water.
Trimethyl-xanthine has another name called theine, and white needle-like crystals has stronger central nervous excitation effect and diuretic properties, is widely used in fields such as medicine, food, makeup.In tealeaves and sheet end sub product 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 technology 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 it is produced the strictness that must obtain relevant department and examines.Common tea-polyphenol extracts or refining enterprise generally is difficult to obtain to produce authorize.Therefore, tea-polyphenol manufacturing enterprise generally can not take recovery measure to theine contained in the production process waste water.Research shows that even the trimethyl-xanthine of low concentration promptly has tangible toxicity to aquatic animals and plants, so trimethyl-xanthine has very big potential destruction to aquatic environment.Because trimethyl-xanthine all shows very strong resistance to soda acid and common oxidizer treatment; Common mikrobe generally also is difficult to degraded and utilizes theine; And be dissolved in theine good dispersivity in the water, can not produce visual impact to water body chroma etc. as other pollutent; Therefore, obtain reflection in the monitoring index that the theine of higher concentration generally also is difficult in BOD, suspended substance and colourity etc. even contain 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 theine waste water.
Summary of the invention
The technical problem that the present invention will solve 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 the 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 present 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 wastewater treatment:
The aerobic treatment stage after anaerobically fermenting is used,
Finishing the back in 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 (promptly 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 to contain the processing of trimethyl-xanthine waste water.The method that bacterial classification is preserved is, in the bacterium liquid numerous product of expansion of absorbancy >=4 of 600 nanometers (promptly), adds the DMSO 99.8MIN. that is equivalent to bacteria liquid long-pending 20%, places subzero 20 degree～subzero 70 degree refrigerators preservations.
Contain in the method for innocent treatment of trimethyl-xanthine waste water in the present invention, pseudomonas putida (fresh weight) is meant: with the above-mentioned bacterium liquid numerous product of expansion of absorbancy >=4 of 600 nanometers (promptly) centrifugal 10-15 minute collected bacterial precipitation under 2000-5000 rev/min condition.
The present invention is applicable to that Wastewater Treated by Activated Sludge Process contains trimethyl-xanthine waste water; Can get involved at the three phases of wastewater treatment and use: both can in coagulation or flocculation back waste water, use; Also can use in the aerobic treatment stage after anaerobically fermenting, also can finish the back and use (see figure 1) in conventional waste water aerobic treatment.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, do not have additional cost basically.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, and conventional aeration adapts to 3-5 days and (promptly after pseudomonas putida is inserted, waste water 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 handling waste water once more afterwards, can contain the harmless treatment of trimethyl-xanthine waste water according to the sewage aerobic treatment method of routine, that is, the treatment time of waste water in aerobic aeration tank (jar) 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 (jar) 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 (jar) was 96 hours; When the trimethyl-xanthine concentration in the waste water surpassed 1500 mg/litre, the treatment time in aerobic aeration tank (jar) was 96～160 hours.Therefore, when trimethyl-xanthine concentration was lower than 500 mg/litre in the waste water, can thoroughly degrade trimethyl-xanthine in the waste water that is equivalent to insert 50,000 times of bacterial classification amounts every day; When trimethyl-xanthine concentration in the waste water during in the 500-1500 mg/litre, can thoroughly degrade trimethyl-xanthine in 2.5 ten thousand times the waste water that is equivalent to insert the bacterial classification amount every day; When trimethyl-xanthine concentration in the waste water surpasses 1500 mg/litre, can the trimethyl-xanthine in the ten thousand times of waste water of 1.5-2.5 that be equivalent to get involved the bacterial classification amount thoroughly 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 isolating pseudomonas putida of tea garden soil, 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 do not have extra input; 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 is applicable to and removes the theine in the theine waste water that contains that produces in tea-polyphenol extraction or the tea-polyphenol treating process.
Description of drawings
Do further explain below in conjunction with the accompanying drawing specific embodiments of the invention.
Fig. 1 is that the waste water conventional activated sludge method that tea-polyphenol production of the present invention and treating process produce is handled and trimethyl-xanthine decomposer (being pseudomonas putida) intervention stage synoptic diagram;
Among the figure: the necessary operation of solid arrow representative, dotted arrow is represented optional operation; Trilateral is represented 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 thoroughly with the original process seamless connection, and does not almost have extra input.
Embodiment
Purifying and the domestication of embodiment 1, pseudomonas putida (being the theine decomposer):
Gather the root topsoil from tea place (being arranged in tea tree germ plasm resource garden, the Zhejiang University Hua Jiachi school district) soil of planting more than 10 years; Remove litter; 1 gram soil is added 1 liter of stirring of sterilized water; Get 0.1 milliliter of supernatant; Be inoculated in be equipped with 20 milliliters with the trimethyl-xanthine be sole carbon source and nitrogenous source the solid bacteria isolation medium (5.0 mmoles/rise Repone K, 1.5 mmoles/rise ferrous sulfate, 1.5 mmoles/rise sal epsom, 0.4 mmole/rise calcium chloride, 0.3 mmole/rise 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, trimethyl-xanthine 25 mmoles/liter, agar 12 grams per liters, in petridish pH7.0); Under 28-30 ℃ of condition, be inverted and cultivated 3 days; Each mono-clonal bacterium bacterial plaque of picking white; Respectively inoculation go into 1 milliliter be the liquid purifying substratum (100 mmoles/rise trimethyl-xanthine do not add agar, and all the other compositions and acidity are with the solid bacteria isolation medium) of sole carbon source and nitrogenous source with the trimethyl-xanthine; 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 of taming (that is pseudomonas putida) so continuously.
The preparation method of above-mentioned solid bacteria isolation medium is following: the trimethyl-xanthine and the 12 gram agar waters 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, 25 mmoles are settled to 1 liter; Use concentration to regulate pH to 7.0 then as the sodium hydroxide of mass concentration 4%; In 121 ℃ of sterilization 20min, get the solid bacteria isolation medium.
The preparation method of aforesaid liquid purifying substratum is following: 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; Using mass concentration then is that 4% sodium hydroxide is regulated pH to 7.0; 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 that 1 liter of pseudomonas putida (Pseudomonas putida) CGMCC NO.5767 is inoculated in 250 liters (5.0 mmoles/rise Repone K, 1.5 mmoles/rise ferrous sulfate, 1.5 mmoles/rise sal epsom, 0.4 mmole/rise calcium chloride, 0.3 mmole/rise 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, trimethyl-xanthine 7.5 mmoles/liter; PH7.0); Under 28-30 ℃, 130-150 rev/min condition, cultivate expand numerous; When substratum during (expand numerous time be about 2-4 days), can be used for containing the processing of trimethyl-xanthine waste water in the absorbancy of 600 nanometers >=4.
The preparation method of the numerous liquid nutrient medium of above-mentioned expansion is: 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, the trimethyl-xanthine 7.5 mmole waters of 5.0 mmoles are settled to 1 liter; Using mass concentration then is that 4% sodium hydroxide is regulated pH to 7.0; 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 handled according to " heterogeneous catalytic oxidation and loop cycle activated sludge process (CASS) combination process are handled 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 bacterium bacterium 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 (jar) that 0.1 kilogram of pseudomonas putida (fresh weight) is inoculated in volume; Behind the waste water behind the feeding anaerobically fermenting; Carry out conventional aeration and adapt to field planting 3 days, promptly after pseudomonas putida is inserted, 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 handling waste water once more afterwards, can contain the harmless treatment of trimethyl-xanthine waste water according to the sewage aerobic treatment method of routine, that is, the treatment time of waste water in aerobic aeration tank (jar) 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 (jar) is 48 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre, 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 (jar) is 96 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre, that is, can thoroughly degrade trimethyl-xanthine in 2.5 tons of waste water 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 (jar) is 96～160 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre; That is, can trimethyl-xanthine in the 1.5-2.5 ton waste water thoroughly 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 bacterium bacterium 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 (jar) that 0.5 kilogram of pseudomonas putida (fresh weight) is inoculated in volume; Behind the waste water behind the feeding anaerobically fermenting; Carry out conventional aeration and adapt to field planting 4 days, promptly after pseudomonas putida is inserted, 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 handling waste water once more afterwards, can contain the harmless treatment of trimethyl-xanthine waste water according to the sewage aerobic treatment method of routine, that is, the treatment time of waste water in aerobic aeration tank (jar) 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 (jar) is 48 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre, 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 (jar) is 96 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre, that is, can thoroughly degrade trimethyl-xanthine in 12.5 tons of waste water 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 (jar) is 96～160 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre; That is, can trimethyl-xanthine in the 7.5-12.5 ton waste water thoroughly 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 bacterium bacterium 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 (jar) that 1 kilogram of pseudomonas putida (fresh weight) is inoculated in volume; Behind the waste water behind the feeding anaerobically fermenting; Carry out conventional aeration and adapt to field planting 5 days, promptly after pseudomonas putida is inserted, 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 handling waste water once more afterwards, can contain the harmless treatment of trimethyl-xanthine waste water according to the sewage aerobic treatment method of routine, that is, the treatment time of waste water in aerobic aeration tank (jar) 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 (jar) is 48 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre, 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 (jar) is 96 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre, 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 surpasses 1500 mg/litre; Treatment time in aerobic aeration tank (jar) is 96～160 hours; Waste water after the processing is through detecting trimethyl-xanthine concentration≤1 mg/litre; That is, can trimethyl-xanthine in the 15-25 ton waste water thoroughly 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 (4)
1. pseudomonas putida; It is characterized in that: 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.
2. the numerous method of the expansion of pseudomonas putida as claimed in claim 1; It is characterized in that: 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.
3. the numerous method of the expansion of pseudomonas putida according to claim 2 is characterized in that:
The numerous liquid nutrient medium of said expansion 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.
4. method for innocent treatment that contains trimethyl-xanthine waste water is characterized in that: contain the pseudomonas putida that 0.8～1.2kg is gone in inoculation in the trimethyl-xanthine waste water per 100 tons, get involved in the stage of following any wastewater treatment and use:
The aerobic treatment stage after anaerobically fermenting is used,
Finishing the back in the waste water aerobic treatment uses;
Said pseudomonas putida is pseudomonas putida (Pseudomonas putida) CGMCC NO.5767.
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| CN106242217A (en) * | 2016-08-26 | 2016-12-21 | 北海运龙环保材料有限责任公司 | Special dose of a kind of anaerobic tank containing zinc |
| CN106277681A (en) * | 2016-08-26 | 2017-01-04 | 北海运龙环保材料有限责任公司 | Special dose of a kind of anaerobic tank containing manganese dioxide |
| CN106277680A (en) * | 2016-08-26 | 2017-01-04 | 北海运龙环保材料有限责任公司 | Special dose of the anaerobic tank of a kind of cupric |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106242219A (en) * | 2016-08-26 | 2016-12-21 | 北海运龙环保材料有限责任公司 | Special dose of the anaerobic tank of a kind of boracic |
| CN106242220A (en) * | 2016-08-26 | 2016-12-21 | 北海运龙环保材料有限责任公司 | Special dose of a kind of anaerobic tank containing magnesium |
| CN106242217A (en) * | 2016-08-26 | 2016-12-21 | 北海运龙环保材料有限责任公司 | Special dose of a kind of anaerobic tank containing zinc |
| CN106277681A (en) * | 2016-08-26 | 2017-01-04 | 北海运龙环保材料有限责任公司 | Special dose of a kind of anaerobic tank containing manganese dioxide |
| CN106277680A (en) * | 2016-08-26 | 2017-01-04 | 北海运龙环保材料有限责任公司 | Special dose of the anaerobic tank of a kind of cupric |
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