CN101935625A - Triethylamine degrading bacterium and microbial agent produced therefrom - Google Patents
Triethylamine degrading bacterium and microbial agent produced therefrom Download PDFInfo
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- CN101935625A CN101935625A CN 201010199254 CN201010199254A CN101935625A CN 101935625 A CN101935625 A CN 101935625A CN 201010199254 CN201010199254 CN 201010199254 CN 201010199254 A CN201010199254 A CN 201010199254A CN 101935625 A CN101935625 A CN 101935625A
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Abstract
The invention provides a degrading bacterium eliminating the pollution of triethylamine and a degrading microbial agent and belongs to the high-technical field of biology. The used strain is a gram staining reaction positive bacterium R4, is determined as arthrobacter protophormiae, and has a biological characteristic of G+. The logarithmic phase thallus is bacilliform, sporeless and aerobic to grow, and can grow by taking the triethylamine as the only carbon source or nitrogen source and thoroughly mineralize the triethylamine to generate ammonia nitrogen. In a flask experiment in a laboratory, the strain can completely degrade 100mg/L triethylamine and solve the problem of the difficult biodegradation of the triethylamine in wastewater treatment.
Description
One, technical field
The invention provides the microbial inoculum of a kind of triethylamine degradation bacteria and production thereof, belong to biological high-tech field, is the pollutent that utilizes in the method for microorganism degrading waste water, is applicable to modern waste water treatment process.
Two, technical background
The organic amine product is widely used in the industries such as agricultural chemicals, medicine, rubber, synthetic dyestuff, oil, process hides, washing composition and organic synthesis, inorganic chemical industry.The organic amine product category is many, and each product all can have remaining raw material and intermediate product to enter in the waste water in process of production, the most high nitrogen low-carbon (LC) of these materials, and poisonous and harmful is difficult to biological degradation, thereby has brought difficulty for the processing of this type of wastewater from chemical industry.
Triethylamine is organic solvent and chemical industry synthetic important source material, the catalyzer of the poly-carbonic acid vinegar phosgenation of conduct in synthetic resins is made, the stopper of tetrafluoroethylene and thiofide; Also can be used as food preservatives, emulsifying agent; Still make the raw material of dyestuff, sterilant simultaneously.Triethylamine (Triethylamine) is a kind of fatty amine, molecular formula (C
2H
5)
3N has intensive ammonia stink, pungency is arranged, inflammable, volatile.Concentration is that the triethylamine of 50mg/L can make the people produce serious lung irritation.
Triethylamine extensively is present in the factory effluents such as organic synthesis, agricultural chemicals, dyestuff, adopts physico-chemical process for the processing that contains triethylamine waste water at present more, and its cost is higher, and easily causes secondary pollution; And bioremediation technology can be degraded and transform because of microorganism itself and had great potential all kinds of different organic pollutants.Therefore, effective bacterial classification that screening obtains removing triethylamine from physical environment is significant.
Three, summary of the invention
Technical problem the objective of the invention is at practical problems in the production practice and demand, develops out a kind of triethylamine degradation bacterial agent, uses the triethylamine of this microbial inoculum in can degrading waste water, makes discharged wastewater met the national standard, reduces simultaneously and produces and use cost.
Be main contents of the present invention below the technical scheme:
The invention provides a kind of degradation bacteria of eliminating triethylamine, its bacterial strain is a strain gram positive bacterium R4, be preserved in Chinese microorganism strain on May 18th, 2010 and preserve management committee common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, deposit number CGMCCNO.3855 is through being accredited as bare glass fly Arthrobacter (Arthrobacter protophormiae).Main biological characteristics is G
+, the logarithmic phase thalline is shaft-like, no gemma, aerobic growth.Can be that sole carbon source, nitrogenous source are grown with the triethylamine, and its thorough mineralising is produced ammonia nitrogen.Shake in the laboratory that the degradation rate to the 100mg/L triethylamine reaches 100% under bottle condition.This bacterium can produce with the general fermentation equipment of fermentation industry.
The technology of using above-mentioned triethylamine degradation bacteria to produce microbial inoculum is: inclined-plane kind-shake-flask seed liquid-seeding tank-product (the packing formulation is liquid bacterial agent).
Detailed implementation step of the present invention is:
1) the test tube kind is inoculated in the LB substratum, shaking culture is to logarithmic phase;
2) above-mentioned cultured bacterial classification is inoculated into 500 liters of seeding tanks by 10% inoculum size, be cultured to logarithmic phase, the used culture medium prescription of seeding tank is: and glucose 1% (mass ratio, Hereinafter the same), (NH
4)
2SO
40.05%, K
2HPO
40.15%, MgSO
40.01%, peptone 0.05%, pH 7.2-7.5;
3) seed liquor is produced a jar cultivation by 10% inoculum size access, it is identical with the seed tank culture base to produce the used substratum of jar;
4) air flow of sterile air is 1 in the culturing process of seeding tank and production jar: 0.6-1.2, stirring velocity is 180-240 rev/min, culture temperature is 30 ℃, the whole process incubation time is 48-60 hour, after the fermentation ends thalline quantity reach 1,000,000,000/more than the ml, the fermentation finish the back nutrient solution go out the jar directly be distributed into liquid agent with plastic barrel or packing bottle.
Beneficial effect
The invention provides a kind of degradation bacteria that triethylamine pollutes of eliminating, laboratory biological degradation experimental result shows, the degradation rate of 100mg/L triethylamine is reached 100%.
It is low, easy to use that the degradation bacterial agent that this invention is produced has a production cost, and the advantage that removal effect is good is adapted at containing in the wastewater treatment of triethylamine and promotes the use of.The present invention is for preserving the ecological environment, and the protection mankind's is healthy, reduces cost for wastewater treatment and has great importance.
Four, description of drawings
Fig. 1 bacterial strain R4 bacterium colony photo
Fig. 2 inoculum size is to the influence of triethylamine degradation efficiency
Fig. 3 temperature is to the influence of triethylamine degradation efficiency
Fig. 4 air flow is to the influence of triethylamine degradation efficiency
Fig. 5 gas chromatographic detection triethylamine degradation effect
Five, embodiment
The separation of 1 bacterial strain and evaluation
Get triethylamine enrichment bacterium liquid, carry out gradient dilution.Diluent is coated the inorganic salt solid medium that contains the 100mg/L triethylamine, and (prescription is: every liter contains 1.5g K
2HPO
4, 0.5g KH
2PO
4, 0.2g MgSO
47H
2O, 1.0g NaCl, 1.0g (NH
4)
2SO
4, 15g agar, pH 7.0) on, cultivate a couple of days for 30 ℃.Therefrom picking list bacterium colony is verified its degradation effect, and the strain bacterial strain that degradation efficiency is higher is preserved, and carries out subsequent experimental.Its bacterium colony picture is seen Fig. 1, is accredited as bare glass fly Arthrobacter (Arthrobacter protophormiae); Called after: R4.Main biological characteristics is G
+, the logarithmic phase thalline is shaft-like, no gemma, aerobic growth.Can be that sole carbon source, nitrogenous source are grown with the triethylamine, and its thorough mineralising is produced ammonia nitrogen.Shake in the laboratory that the degradation rate to the 100mg/L triethylamine reaches 100% (Fig. 5) under bottle condition.This bacterium can produce with the general fermentation equipment of fermentation industry.
2 laboratory biological degradation experiments
2.1 inoculum size is to the influence of triethylamine degradation efficiency
R4 is cultured to the logarithm later stage in the LB substratum, the inoculum size by 0.1,0.5,1,2,3,5% inserts the minimal medium that contains the 100mg/ triethylamine respectively, and (the minimal medium prescription is: every liter contains 1.5g K
2HPO
4, 0.5gKH
2PO
4, 0.2g MgSO
47H
2O, 1.0g NaCl, 1.0g (NH
4)
2SO
4, pH 7.0), Hereinafter the same) in, 30 ℃, 160r/min shaking table are cultivated, the content of sampling and measuring triethylamine after 20 hours, result such as Fig. 2.Measurement result shows, arranges from low to high by inoculum size, and the degradation rate of furans pellet is followed successively by 10%, 38%, 65%, 80%, 90%, 91%.This shows that the size of inoculum size has direct relation to the degradation efficiency of triethylamine, when inoculum size less than 3% the time, improve inoculum size, the degradation efficiency of triethylamine has significantly raising, but inoculum size reaches at 5% o'clock, degradation rate has only trace to improve, and enzyme reaction at this moment is near saturated.
2.2 temperature is to the influence of triethylamine degradation efficiency
R4 cultivates the logarithm later stage in the LB substratum, receive in the minimal medium that contains the 100mg/ triethylamine by 1% inoculum size, place respectively under 15,20,25,30,37 ℃ the temperature, the 160r/min shaking table is cultivated, measure the wherein content of triethylamine behind the 28h, the result as shown in Figure 3.In the time of 15 ℃, R4 is 14% to the degradation rate of triethylamine; In 20 ℃～30 ℃ scope, along with the red performance of the rising degradation bacteria of temperature degraded furans obviously rises, in the time of 30 ℃, degradation rate reaches 96%.And when temperature reached 37 ℃, R4 descended to the degradation rate of triethylamine, only was 9%.Above data declaration, the suitableeest degradation temperature of R4 is 30 ℃; Under lower temperature, still has certain degradation capability; And the low or too high envrionment temperature of mistake can suppress the biological degradation of triethylamine.
2.3 air flow is to the influence of triethylamine degradation efficiency
In the triangular flask of 5 250mL, be respectively charged into 30,50,70,100,150mL contains in the minimal medium of 100mg/ triethylamine, recently indicates air flow with dress liquid percentage.R4 cultivates the logarithm later stage in the LB substratum, and the inoculum size by 1% is inoculated in above-mentioned each substratum, 30 ℃, the cultivation of 160r/min shaking table, the degradation rate of mensuration triethylamine after 20 hours, result such as Fig. 4.20 hours measurement result show be equipped with 30,50,70,100, the degradation rate of furans pellet is respectively 91%, 86%, 79%, 65% and 42% in the triangular flask of 150mL nutrient solution, the process of R4 degraded triethylamine is an aerobic processes, and big air flow helps the degraded of triethylamine.
2.4 R4 is to the tolerance concentration test of triethylamine
R4 is cultured to the logarithm later stage in the LB substratum, in the minimal medium by 2% triethylamine that is seeded to different concns (100,200,500,1000mg/L), measure the content of triethylamine at regular intervals, result such as table 1.Growth does not produce inhibition to 1000mg/L high density triethylamine to R4, and the 100mg/L triethylamine of degrading fully needs 32h.
Table 1 triethylamine initial concentration is to the influence of R4 degraded
3 produce enforcement
The original seed of microorganism triethylamine degradation bacteria R4 of the present invention is activated on culture dish, and measure degradation property, be inoculated on the test tube slant standby.The test tube kind is inoculated in the 1000ml that contains 200ml LB substratum (LB culture medium prescription g/L: yeast extract paste 5.00, peptone 10.00, NaCl 10.00, pH 7.0) and shakes in the bottle, and constant-temperature shaking culture is prepared the inoculation seeding tank to logarithmic phase.500 liters of seeding tanks, 400 liters of charging capacitys, culture medium prescription is: glucose 1%, (NH
4)
2SO
40.05%, K
2HPO
40.15%, MgSO
40.01%, peptone 0.05%, pH 7.2-7.5.The back 115 ℃ of high pressure moist heat sterilizations that finish that feed intake, be cooled to 30 ℃ after, the above-mentioned cultured bottle bacterial classification that shakes is inserted 500 liters of seeding tanks by 10% inoculum size, be cultured to logarithmic phase, stirring velocity is 220 rev/mins, sterile air feeding amount is 1: 0.8.The seed liquor that will reach logarithmic phase is produced a jar cultivation by 10% inoculum size access, and it is identical with the seed tank culture base to produce the used medium component of jar.Produce 5 tons of tankages, 4.5 tons of charging capacitys.A back production jar 1.1kg/cm feeds intake
2Pressure under, 115 ℃ of high pressure moist heat sterilizations, below the sterilization postcooling to 30 ℃, logical sterile air keeps sterile state standby.A postvaccinal production jar temperature is controlled at 30 ℃, and the air flow of sterile air is 1 in the culturing process of production jar: 0.8-1.0, and stirring velocity is 240 rev/mins, the whole process flow incubation time is 48-60 hour.After the fermentation ends thalline quantity reach 1,000,000,000/more than the ml.
The fermentation finish the back nutrient solution go out the jar directly be distributed into liquid dosage form with plastic barrel or packing bottle.
Claims (2)
1. triethylamine degradation bacteria, it is characterized in that this microorganism is the bacterial strain R4 of the bare glass fly Arthrobacter (Arthrobacter protophormiae) of Gram-positive, be preserved in Chinese microorganism strain on May 21st, 2010 and preserve management committee common micro-organisms center, deposit number CGMCC NO.3855.Main biological characteristics is G
+, the logarithmic phase thalline is shaft-like, no gemma, aerobic growth.Can be that sole carbon source, nitrogenous source are grown with the triethylamine, and its thorough mineralising is produced ammonia nitrogen.Shake in the laboratory that the degradation rate to the 100mg/L triethylamine reaches 100% under bottle condition.
2. degradation bacterial agent of producing with the described triethylamine degradation bacteria of claim 1 is to produce by the following method:
1) the test tube kind is inoculated in the LB substratum, shaking culture is to logarithmic phase;
2) above-mentioned cultured bacterial classification is inoculated into 500 liters of seeding tanks by 10% inoculum size, be cultured to logarithmic phase, the used culture medium prescription of seeding tank is: and glucose 1% (mass ratio, Hereinafter the same), (NH
4)
2SO
40.05%, K
2HPO
40.15%, MgSO
40.01%, peptone 0.05%, pH 7.2-7.5;
3) seed liquor is produced a jar cultivation by 10% inoculum size access, it is identical with the seed tank culture base to produce the used substratum of jar;
The air flow of sterile air is 1 in the culturing process of seeding tank and production jar: 0.6-1.2, stirring velocity is 180-240 rev/min, culture temperature is 30 ℃, the whole process incubation time is 48-60 hour, after the fermentation ends thalline quantity reach 1,000,000,000/more than the ml, the fermentation finish the back nutrient solution go out the jar directly be distributed into liquid agent with plastic barrel or packing bottle.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN103602619A (en) * | 2013-11-21 | 2014-02-26 | 南京理工大学 | Thauera sp. capable of degrading triethylamine as well breeding method and application of Thauera sp. |
| CN103849584A (en) * | 2014-01-15 | 2014-06-11 | 山东新时代药业有限公司 | Triethylamine degrading bacterium and application thereof |
| CN106497837A (en) * | 2016-11-03 | 2017-03-15 | 淮阴师范学院 | Gold orange II degradation bacterias AO7 2 and its microbial inoculum of production |
| CN115141782A (en) * | 2022-08-16 | 2022-10-04 | 浙江省农业科学院 | Sulfamethazine degrading microbial inoculum and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1807585A (en) * | 2005-12-23 | 2006-07-26 | 南京农业大学 | Highly effective phosphorus removal bacteria and its produced bacteria formulation |
-
2010
- 2010-06-12 CN CN2010101992542A patent/CN101935625B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1807585A (en) * | 2005-12-23 | 2006-07-26 | 南京农业大学 | Highly effective phosphorus removal bacteria and its produced bacteria formulation |
Non-Patent Citations (3)
| Title |
|---|
| 《Arch Microbiol》 19941231 J.Frings et al Fermentatative degradation of triethanolamine by a homoacetogenic bacterium 全文 1，2 第162卷, 2 * |
| 《江西农业学报》 20091231 吴守中等 反硝化聚磷菌C18脱氮除磷特性研究 全文 1，2 第21卷, 第10期 2 * |
| 《环境工程》 20081231 陈立伟等 高效聚磷菌TP16强化除磷 全文 1，2 第26卷, 第6期 2 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN103243057B (en) * | 2013-05-27 | 2015-03-25 | 江苏南资环保科技有限公司 | Pseudomonas SYA-1 for degrading triethylamine and application of pseudomonas SYA-1 |
| CN103602619A (en) * | 2013-11-21 | 2014-02-26 | 南京理工大学 | Thauera sp. capable of degrading triethylamine as well breeding method and application of Thauera sp. |
| CN103602619B (en) * | 2013-11-21 | 2015-09-30 | 南京理工大学 | The Soxhlet bacterium of one strain degradable triethylamine, selection and application |
| CN103849584A (en) * | 2014-01-15 | 2014-06-11 | 山东新时代药业有限公司 | Triethylamine degrading bacterium and application thereof |
| CN103849584B (en) * | 2014-01-15 | 2016-05-25 | 山东新时代药业有限公司 | One strain triethylamine degradation bacteria and application thereof |
| CN106497837A (en) * | 2016-11-03 | 2017-03-15 | 淮阴师范学院 | Gold orange II degradation bacterias AO7 2 and its microbial inoculum of production |
| CN106497837B (en) * | 2016-11-03 | 2019-04-02 | 淮阴师范学院 | Gold orange II degradation bacteria AO7-2 and its microbial inoculum of production |
| CN115141782A (en) * | 2022-08-16 | 2022-10-04 | 浙江省农业科学院 | Sulfamethazine degrading microbial inoculum and preparation method and application thereof |
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| CN101935625B (en) | 2012-05-09 |
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