CN103849584B - One strain triethylamine degradation bacteria and application thereof - Google Patents
One strain triethylamine degradation bacteria and application thereof Download PDFInfo
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Abstract
The invention belongs to biological technical field, be specifically related to strain triethylamine degradation bacteria and an application thereof. Triethylamine degradation bacteria T-5 of the present invention belongs to Bacillus cercus (Bacillus), is preserved in Chinese Typical Representative culture collection center, and preservation date is on November 21st, 2013, does is deposit number CCTCC? NO:M2013588. Described bacterial strain can utilize triethylamine to carry out growth and breeding as carbon source, nitrogenous source, can be in the shaking flask that contains 4000mg/L triethylamine growth and breeding, be 100% to degradation rate in 1500mg/L triethylamine 72h. To containing in the processing of high concentration triethylamine industrial wastewater, in the time that triethylamine concentration is less than 2000mg/L, the clearance of triethylamine is reached to 100%, COD clearance and improve 30～40%. This bacterial strain can be applicable to the biological reinforced processing of pharmacy waste water, pesticide processing waste water, has a extensive future.
Description
Technical field
The invention belongs to biological technical field, be specifically related to strain triethylamine degradation bacteria and an application thereof.
Background technology
Triethylamine is the synthetic important source material of organic solvent and chemical industry, the poly-carbonic acid vinegar phosgene of conduct in synthetic resin is manufacturedCatalyst, the polymerization inhibitor of tetrafluoroethene and thiofide; Also can be used as food preservative, emulsifying agent; Still simultaneouslyManufacture the raw material of fuel, pesticide. Triethylamine is a kind of aliphatic amine, molecular formula (C2H5)3N, has strong degree ammonia stink, hasExcitant, inflammable, volatile.
Triethylamine is extensively present in the factory effluents such as organic synthesis, agricultural chemicals, pharmacy, fuel, at present for containing triethylamineThe processing of waste water adopts physico-chemical process more. Compared with physico-chemical process, it is large that biological treatment has treating capacity, costLow, mild condition, does not produce the features such as secondary pollution, more meets the requirement of the friendly type of constructing environment and conservation-minded society.But traditional biologic treating technique is difficult to meet the environmental requirement of increasingly stringent. In recent years, biological reinforcing technology is not changingOn the basis of existing treatment facility, by the interpolation of microorganisms with specific functions, improve original biological treatment system to target stainsThe removal efficiency of thing, has improved sewage treatment capacity greatly. Chinese invention patent CN101935625A discloses a kind of triethylamineThe microbial inoculum of degradation bacteria and production thereof, this bacterium only, for laboratory shaking flask, does not relate to practical application in industry. Therefore, from natural ringIn the processing that in border, screening obtains can removing the efficient bacterium of triethylamine and being applied to industrial wastewater, there is great meaningJustice.
Summary of the invention
For the triethylamine of effectively degrading, the invention discloses a strain triethylamine degradation bacteria, and be successfully applied to containIn the industrial wastewater of triethylamine.
One of object of the present invention is to disclose the bacterial strain T-5 of a high-efficiency degradation triethylamine.
Triethylamine degradation bacteria T-5 of the present invention belongs to Bacillus cercus (Bacilluscereus.FranklandT-5) be preserved in Chinese Typical Representative culture collection center (ChinaCenterforTypeCultureCollection, letterClaim CCTCC), deposit number CCTCCNo:M2013588 preservation date on November 21st, 2013.
The 16SrDNA sequence of bacterial strain T-5 of the present invention is:
GATGGATTAAGAGCTTGCTCTTATGAAGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCATAAGACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATAACATTTTGAACCGCATGGTTCGAAATTGAAAGGCGGCTTCGGCTGTCACTTATGGATGGACCCGCGTCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCAACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTAGGGAAGAACAAGTGCTAGTTGAATAAGCTGGCACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGAATTATTGGGCGTAAAGCGCGCGCAGGTGGTTTCTTAAGTCTGATGTGAAAGCCCACGGCTCAACCGTGGAGGGTCATTGGAAACTGGGAGACTTGAGTGCAGAAGAGGAAAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGAGATATGGAGGAACACCAGTGGCGAAAGCGACTTTCTGGTCTGTAACTGACACTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAAAGGGTTTCCTGCCTTTAGTGCTGAAGTTAACGCATTAAGCACTCCGCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGCCCGCACAAGCGTGAG
Above-mentioned 16SrDNA portion gene sequence (being SEQIDNO.1) is to send out in KF877717 with GenBank accession numberThe 16SrDNA sequence of the Bacillus cercus (Bacilluscereus) of table is carried out homology comparison, and similarity reaches 99%, nameFor Bacillus cercus T-5(Bacilluscereus.FranklandT-5).
Triethylamine degradation bacteria T-5 of the present invention also has following character:
Described bacterial strain T-5 is gram-positive bacteria;
Described bacterial strain T-5 is pink through cultivating the bacterium colony of gained, and neat in edge is rounded, smooth moistening, middle cardiac prominenceRise;
The preference temperature of described bacterial strain T-5 growth is that 25～35 DEG C, appropriate pH are 5.5～7.5.
Those skilled in the art can cultivate triethylamine degradation bacteria T-5 of the present invention according to conventional method, butThat more preferred cultural method is: bacterial strain T-5 is inoculated in certain volume proliferated culture medium, 25～35 DEG C, pH are 5.5～Under 7.5 condition, shaking table vibration 3 days, obtains T-5 bacteria suspension, and thalline quantity is 109Individual/mL; By T-5 bacterial suspension inoculation in propagationCulture medium expands cultivation, and condition of culture is the same; T-5 bacteria suspension is coated to solid medium flat board, after bacterium colony grows,Be placed under 4 DEG C of conditions and preserve as bacterial classification. Proliferated culture medium composition described above is: KH2PO4，0.09；K2HPO4，0.22；NaH2PO4，0.26；MgSO4·7H2O，0.23；CaCl2，0.28；FeCl3, 0.003; Triethylamine, 1.5(unit: g/L). DescribedSolid medium composition be: KH2PO4，0.09；K2HPO4，0.22；NaH2PO4，0.26；MgSO4·7H2O，0.23；CaCl2，0.28；FeCl3, 0.003; Triethylamine, 1.0; Agar, 15～20. (unit: g/L).
Bacterial strain T-5 of the present invention can carry out using triethylamine as Carbon and nitrogen sources growth and breeding and by its degraded:
In solid medium, KH2PO4、K2HPO4、NaH2PO4、MgSO4·7H2O、CaCl2、FeCl3Deng inorganic salts compositionFor maintaining the required osmotic pressure of strain growth, agar is only as the carrier of strain growth, and triethylamine is that strain growth breeding is carriedFor Carbon and nitrogen sources.
T-5 bacteria suspension is coated to the solid medium flat board that triethylamine concentration is 4000mg/L, and visible colony growth is numerousGrow. See Fig. 1. T-5 bacteria suspension is inoculated in to the proliferated culture medium that 200mL triethylamine concentration is 1500mg/L by 1% inoculum concentrationIn, 25～35 DEG C of constant temperature culture, pH is 5.5～7.5, and rotating speed is 150rpm, and this bacterium is degradable by it in 72h. Detect knotFruit sees Fig. 2 and Fig. 3.
Two of object of the present invention is this bacterial strain T-5 for the treatment of the industrial wastewater that contains triethylamine.
In the present invention, the assay method of triethylamine is gas chromatography (GB/T23964-2009), described COD assay methodFor potassium dichromate method (GB11914-89).
T-5 bacteria suspension is inoculated in the waste water that 200mL triethylamine concentration is 1500mg/L to 25～35 by 1% inoculum concentrationDEG C constant temperature culture, pH is 5.5～7.5, and rotating speed is 150rpm, and this bacterium is degradable by it in 72h.
T-5 bacteria suspension is added to and in activated Sludge System, carries out biological reinforcedly by different dosages, and service condition: pH is5.5～7.5, temperature is 25～35 DEG C, and dissolved oxygen is 3.0～4.0mg/L, and triethylamine concentration is 1400～2000mg/L, operationMode: water inlet, aeration, stops aeration and sedimentation, and draining is a cycle of operation, and hydraulic detention time 2～3 days adds bacterium for every 2 days outstandingLiquid 1 time, adds 2 months and after stable effluent quality, stops adding, system continuous service. Control group is set simultaneously, and control group is notAdd bacteria suspension, other methods of operation are identical with biological reinforced group, results of regular determination water outlet triethylamine content and COD.
The present invention compared with prior art, has following outstanding technique effect:
Bacterial strain of the present invention can utilize triethylamine to carry out growth and breeding as Carbon and nitrogen sources, can containGrowth and breeding in the laboratory shaking flask of 4000mg/L triethylamine, under the shaking flask condition of laboratory in 1500mg/L triethylamine 72hDegradation rate is 100%. To containing in the processing of high concentration triethylamine industrial wastewater, in the time that triethylamine concentration is less than 2000mg/L, rightThe clearance of triethylamine reaches 100%, COD clearance and improves 30～40%.
Described bacterial strain can be applicable to the biological reinforced processing of pharmacy waste water, pesticide processing waste water, passes through the embodiment of the present inventionChecking known: the detection of biological strengthening group system continuous service water outlet testing result of 4 months is that the clearance of triethylamine is reachedReach 90% to 100%, COD clearance.
Described bacterial strain is added in activated Sludge System can reach Screening of high efficient paracetamolum, applies at present in Industrial Wastewater TreatmentActivated Sludge System operation process is sequencing batch active sludge (being called for short SBR) and Cyclic Activated Sludge System (letter the most widelyClaim CASS). SBR technique has that sludge settling is effective, output is few, does not need to arrange second pond, adapts to water quality and quantity variation etc. excellentPoint, cyclic activated sludge system has increased biological selecting area, has strengthened the hydrolysis to hardly degraded organic substance. Described bacterial strain is added toState and in two kinds of activated Sludge System, have that bacterial classification is adding less, tolerance triethylamine concentration advantages of higher, be applicable to the work containing triethylamineThe processing of industry waste water.
In addition, it is large that biological treatment of the present invention has treating capacity, and cost is low, and mild condition does not produce secondary dirtThe feature such as dye, be especially applicable to the requirement of the friendly type of constructing environment and conservation-minded society, application prospect is boundless.
Brief description of the drawings
Fig. 1 .T-5 is at the solid medium colonial morphology of 4000mg/L triethylamine concentration;
Fig. 2. triethylamine content in original proliferated culture medium;
Fig. 3. triethylamine content in proliferated culture medium after T-5 processes 72h.
Detailed description of the invention
Now further describe beneficial effect of the present invention by following examples, embodiment is only for the object of illustration,Do not limit the scope of the invention those of ordinary skill in the art make according to the present invention simultaneously apparent change and modificationWithin being also contained in the scope of the invention.
Embodiment 1: the cultural method of triethylamine degradation bacteria T-5
Bacterium T-5 of the present invention is inoculated in 10mL proliferated culture medium, under the condition that is 7.5 at 25 DEG C, pH, is placed in perseveranceIn temperature vibration shaking table, vibrate 3 days, obtain T-5 bacteria suspension, thalline quantity is 109Individual/mL. T-5 bacteria suspension is coated to solid trainingSupport base flat board, after bacterium colony grows, be placed under 4 DEG C of conditions and preserve as bacterial classification.
Wherein, described proliferated culture medium composition is: KH2PO4，0.09；K2HPO4，0.22；NaH2PO4，0.26；MgSO4·7H2O，0.23；CaCl2，0.28；FeCl3, 0.003; Triethylamine, 1.5(unit: g/L).
Described solid medium composition is: KH2PO4，0.09；K2HPO4，0.22；NaH2PO4，0.26；MgSO4·7H2O，0.23；CaCl2，0.28；FeCl3, 0.003; Triethylamine, 1.0; Agar, 15～20. (unit: g/L).
Embodiment 2: the expansion cultural method of triethylamine degradation bacteria T-5
Get T-5 bacteria suspension 2mL in embodiment 1 and be inoculated in 200mL proliferated culture medium and expand and cultivate, at 35 DEG C, pH beUnder 5.5 condition, be placed in constant temperature oscillation shaking table and vibrate 3 days, obtain T-5 bacteria suspension, thalline quantity is 109Individual/mL. Get above-mentionedT-5 bacteria suspension 20mL is inoculated in 2000mL proliferated culture medium to expand and cultivates, and under the condition that is 7.0, is placed in constant temperature at 30 DEG C, pHIn vibration shaking table, vibrate 3 days, obtain T-5 bacteria suspension, thalline quantity reaches 109Individual/mL.
The shake flat experiment of embodiment 3:T-5 to 1500mg/L waste water
Get the cultured T-5 bacteria suspension of embodiment 1 2mL and be inoculated in the waste water that 200mL triethylamine concentration is 1500mg/L,30 DEG C of constant temperature culture, pH is 7.0, reacts 72 hours. After getting the centrifugal 5min of 10mL bacteria suspension 3000rpm, get supernatant mensuration, notDetect triethylamine.
Embodiment 4:T-5 is biological reinforced to agricultural chemicals waste water
Get Shandong insecticide factory factory effluent, waste water quality is: triethylamine 96354mg/L, COD84432mg/L. By above-mentionedWastewater dilution to triethylamine concentration is after 1400mg/L, COD concentration are 1200mg/L, get 30L join 40L with SBR techniqueIn the activated Sludge System of operation, get the cultured T-5 bacteria suspension of embodiment 2 150mL and be added in above-mentioned activated Sludge SystemCarry out biological reinforced. Service condition: pH is 5.5, and temperature is 35 DEG C, and dissolved oxygen is 3.0mg/L, the method for operation: water inlet, 1h, exposes to the sunGas 12h, precipitation 4h, draining 1h is one-period, hydraulic detention time 2 days adds bacteria suspension 1 time, adds at every turn for every 2 days150mL, stopped adding after 2 months. Control group is set simultaneously, does not add bacteria suspension, other methods of operation and biological reinforced group of phaseWith, the clearance of detection of biological strengthening group and control group triethylamine and COD, the system continuous service water outlet testing result of 4 monthsAs table 1.
Table 1 adopts biological reinforcing method to the dense triethylamine water treatment effect of height
Embodiment 5:T-5 is biological reinforced to pharmacy waste water
Get Shandong pharmaceutical factory factory effluent, waste water quality is: triethylamine 68316mg/L, COD51432mg/L. By above-mentionedWastewater dilution to triethylamine concentration is after 2000mg/L, COD concentration are 1500mg/L, get 500L join 550L with CASS workIn the activated Sludge System of skill operation, get the cultured T-5 bacteria suspension of embodiment 2 1500mL and be added to above-mentioned activated Sludge SystemIn carry out biological reinforced. Service condition: pH is 7.5, and temperature is 25 DEG C, and dissolved oxygen is 4.0mg/L, the method for operation: water inlet aeration2h, aeration 12h, sedimentation and drawing 4h is one-period, hydraulic detention time 3 days adds bacteria suspension 1 time, adds at every turn for every 2 days1500mL, stopped adding after 2 months. Control group is set simultaneously, does not add bacteria suspension, other methods of operation and biological reinforced group of phaseWith, the clearance of detection of biological strengthening group and control group triethylamine and COD, the system continuous service water outlet testing result of 4 monthsAs table 2.
Table 2 adopts biological reinforcing method to the dense triethylamine water treatment effect of height
Claims (11)
1. a strain triethylamine degradation bacteria T-5, belongs to Bacillus cercus (Bacilluscereus), is preserved in Chinese Typical Representative micro-Biological preservation center, deposit number is CCTCCNO:M2013588.
2. triethylamine degradation bacteria T-5 as claimed in claim 1, is characterized in that, described bacterial strain T-5 can utilize triethylamine as carbonSource and nitrogenous source carry out growth and breeding.
3. the application of triethylamine degradation bacteria T-5 in triethylamine degraded as claimed in claim 1 or 2.
4. the application in the industrial wastewater that triethylamine degradation bacteria T-5 contains triethylamine in processing as claimed in claim 1 or 2.
5. application as claimed in claim 4, is characterized in that, described industrial wastewater is that pharmacy waste water or pesticide processing are uselessWater.
6. application as claimed in claim 5, is characterized in that, the triethylamine concentration in described industrial wastewater is less than 4000mg/L。
7. application as claimed in claim 5, is characterized in that, the triethylamine concentration in described industrial wastewater is less than 2000mg/L。
8. application as claimed in claim 5, is characterized in that, the Industry Waste that adopts biological reinforcing method processing to contain triethylamineWater.
9. application as claimed in claim 8, is characterized in that, described biological reinforcing method for to add in activated Sludge SystemDescribed t bacteria-5.
10. application as claimed in claim 9, is characterized in that, described dosing method is to add once for every two days, adds continuouslyTwo months.
11. application as claimed in claim 9, is characterized in that: described activated Sludge System is CASS system or SBR system.
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| CN101781024A (en) * | 2009-12-01 | 2010-07-21 | 吉林大学 | Composite microbial inoculum for quickly degrading nitrobenzene and phenylamine, and preparation method and application thereof |
| CN101935625A (en) * | 2010-06-12 | 2011-01-05 | 南京农业大学 | Triethylamine degrading bacterium and microbial agent produced therefrom |
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Patent Citations (3)
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| CN101781024A (en) * | 2009-12-01 | 2010-07-21 | 吉林大学 | Composite microbial inoculum for quickly degrading nitrobenzene and phenylamine, and preparation method and application thereof |
| CN101935625A (en) * | 2010-06-12 | 2011-01-05 | 南京农业大学 | Triethylamine degrading bacterium and microbial agent produced therefrom |
| CN103243057A (en) * | 2013-05-27 | 2013-08-14 | 江苏南资环保科技有限公司 | Pseudomonas SYA-1 for degrading triethylamine and application of pseudomonas SYA-1 |
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