CN105199992B - The composite bacteria agent and its preparation method and application of aniline and ammonia nitrogen is removed simultaneously - Google Patents
The composite bacteria agent and its preparation method and application of aniline and ammonia nitrogen is removed simultaneously Download PDFInfo
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- CN105199992B CN105199992B CN201510681384.2A CN201510681384A CN105199992B CN 105199992 B CN105199992 B CN 105199992B CN 201510681384 A CN201510681384 A CN 201510681384A CN 105199992 B CN105199992 B CN 105199992B
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- aniline
- composite bacteria
- bacteria agent
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Abstract
Composite bacteria agent and its preparation method and application that is a kind of while removing aniline and ammonia nitrogen, the composite bacteria agent are respectively by the pseudomonad Z1 and acinetobacter calcoaceticus Y1 that prepare by 1:1 volume ratio is combined, the composite bacteria agent being combined is inoculated in aniline waste water, in 48h, the degradation rate of aniline is 93.54%, and COD degradation rate is 88.41%, ammonia nitrogen surplus is 4.39mg/L, effectively eliminates the ammonia nitrogen that aniline and aniline accumulate in degradation process simultaneously.
Description
Technical field
The present invention relates to a kind of composite bacteria agents and its preparation method and application of removal aniline and ammonia nitrogen, especially by false single
Born of the same parents bacterium Z1 and acinetobacter calcoaceticus Y1 be combined it is a kind of for and meanwhile remove the ammonia nitrogen of release during aniline and degrading aniline
Composite bacteria agent and its preparation method and application.
Background technology
Aniline is colourless one kind, oily, flammable and high toxicity liquid, is important Organic Chemicals, is widely used
It is that a kind of serious pollution environment is harmful to what is be detrimental to health in industries such as national defence, printing and dyeing, plastics, paint, pesticide and medicine
Substance.In recent years, it reaches its maturity to the research of aniline waste water processing, the method for removal Aniline type organic is substantially divided
For：Physical, chemical method and bioanalysis.But aniline can release ammonia nitrogen in degradation process, and ammonia nitrogen is the synthesis of water quality monitoring
One of index, ammonia nitrogen in high density can cause body eutrophication, threaten ecological environment and health.Therefore, it is necessary to consider
Except the ammonia nitrogen discharged during aniline degradation, the discharge of pollutant and by-product is reduced to the greatest extent, this not only has practical application
Reference value, it is also significant to water resource protection.
Due to the complexity of pollutant structure and the diversity of type, multiple-microorganism is selected to form a biosystem,
It allows each bacterium therein to play its best-of-breed functionality, and is accorded with come high efficiency degradation of contaminant using the synergistic effect between microorganism
Close the needs of practice.Since degrading aniline bacterium releases a large amount of ammonia nitrogens during removing aniline, and allotrophic nitrobacteria
Have the function of effective degradation of ammonia nitrogen under the conditions of high C/N again, composite bacteria agent is made in this two classes bacterium, can effectively be gone simultaneously
The ammonia nitrogen accumulated in degradation process except aniline and aniline.
Invention content
The object of the present invention is to provide a kind of while removing the composite bacteria agent and its preparation method and application of aniline and ammonia nitrogen,
The ammonia nitrogen that the composite bacteria agent can simultaneously effective remove the aniline of serious pollution environment and be accumulated during degrading aniline.
The purpose of the present invention is what is be achieved through the following technical solutions.
Composite bacteria agent that is a kind of while removing aniline and ammonia nitrogen, the composite bacteria agent is by microphenomenon of degradation of aniline and heterotrophic nitrification
Bacterium is compound and constitutes；Wherein：
Microphenomenon of degradation of aniline belongs to pseudomonas (Pseudomonas), is named as：Pseudomonas sp.Z1；Preservation
Registration number is：CGMCC No.11444；Preservation mechanism is：China Committee for Culture Collection of Microorganisms's common micro-organisms center
(CGMCC)；The preservation time is：On September 24th, 2015；
Allotrophic nitrobacteria belongs to acinetobacter (Acinetobacter)；It is named as：Acinetobacter
sp.Y1；Preservation registration number is：CGMCC No.6563；Preservation mechanism is：China Committee for Culture Collection of Microorganisms is common
Microorganism center (CGMCC)；The preservation time is：On September 14th, 2012.
Wherein, the ratio between the bacterium solution volume of pseudomonad Z1 and acinetobacter calcoaceticus Y1 are 1:1.
A kind of preparation method for composite bacteria agent that is above-mentioned while removing aniline and ammonia nitrogen, the preparation method is following
What step carried out：
(1) preparation method of pseudomonad Z1
It is inoculated into 100mL fluid nutrient mediums from a small amount of thalline of picking on the inclined-plane of 4 DEG C of preservations, at 120rpm, 30 DEG C
Shaken cultivation 18h, until bacteria suspension OD600Stop culture when value is 0.6~0.8, obtains pseudomonad Z1 bacterium solutions；The vacation unit cell
The medium component of bacterium Z1 is：KH2PO41.0g/L；K2HPO41.0g/L；NaCl 1.0g/L；MgSO40.005g/L；
MnSO40.0017g/L；H3BO30.00116g/L；ZnSO4·7H2O 0.00115g/L；FeSO4·7H2O 0.003g/L；Aniline
0.4g/L；PH value is 7.0-7.2；
(2) preparation method of acinetobacter calcoaceticus Y1
It is inoculated into 100mL fluid nutrient mediums from a small amount of thalline of picking on the inclined-plane of 4 DEG C of preservations, at 120rpm, 30 DEG C
Shaken cultivation 14h, until bacteria suspension OD600Stop culture when value is 1.0~1.2, obtains acinetobacter calcoaceticus Y1 bacterium solutions；The not lever
The medium component of bacterium Y1 is：Sodium citrate, 4.902g/L；(NH4)2SO4, 0.472g/L；MgSO4, 0.05g/L；K2HPO4,
0.2g/L；NaCl, 0.12g/L；MnSO4, 0.01g/L；FeSO4·7H2O, 0.01g/L；PH value is 7.0-7.2；
(3) the pseudomonad Z1 and acinetobacter calcoaceticus Y1 prepared above-mentioned steps presses 1:1 volume ratio is mixed, and is obtained micro-
Biological compound fungi agent.
A kind of to be used for application that is above-mentioned while removing aniline and ammonia nitrogen composite bacteria agent, the application is micro- by what is prepared
Biological compound fungi agent is used for while removing the ammonia nitrogen that aniline and its aniline accumulate in degradation process.
Wherein, described at the same remove the ammonia nitrogen that aniline and its aniline accumulate in degradation process be by inoculum concentration be 4%
Composite bacteria agent is inoculated in aniline waste water by volume ratio.
It is described composite bacteria agent is inoculated in aniline waste water be by pseudomonad Z1 and acinetobacter calcoaceticus Y1 by volume be 1:1
It is inoculated in aniline waste water, composite bacteria agent while aniline degradation and ammonia nitrogen, after 48h, degradation rate of aniline is 93.54%, COD degradation
Rate is 88.41%, and ammonia nitrogen surplus is 4.39mg/L.
Realize above-mentioned provided composite bacteria agent that is a kind of while removing aniline and ammonia nitrogen of the present invention and preparation method thereof and
Using compared with prior art, composite bacteria agent described in this method is by pseudomonad (Pseudomonas sp.) Z1CGMCC
No.11444 and acinetobacter calcoaceticus (Acinetobacter sp.) Y1CGMCC No.6563 presses 1:The ratio between 1 bacterium solution volume it is compound and
At, composite bacteria agent is inoculated in aniline waste water by the volume ratio of inoculum concentration 4%, at this point, composite bacteria agent simultaneously aniline degradation and
The resultant effect of ammonia nitrogen is best, and after 48h, degradation rate of aniline 93.54%, COD degradation rate is 88.41%, and ammonia nitrogen surplus is
4.39mg/L。
Description of the drawings
Fig. 1 is the comparison diagram of the degrading aniline effect and Z1 aniline degradation effects of this composite bacteria agent.
Fig. 2 is the comparison diagram of the ammonia nitrogen accumulation effect and Z1 accumulation ammonia nitrogen amounts of this composite bacteria agent.
Fig. 3 is the comparison diagram of the COD degradation effect and Z1 degradation COD effects of this composite bacteria agent.
Specific implementation mode
It illustrates below the specific implementation mode further illustrated the present invention, but the protection model being not intended to limit the present invention
It encloses.
Specific implementation mode 1
Implement composite bacteria agent that is a kind of while removing aniline and ammonia nitrogen, which is by microphenomenon of degradation of aniline and heterotrophism nitre
Change bacterium to be combined.
Microphenomenon of degradation of aniline belongs to pseudomonas (Pseudomonas), is named as：Pseudomonas sp.Z1；It protects
Hiding registration number is：CGMCC No.11444；Preservation mechanism is：In China Committee for Culture Collection of Microorganisms's common micro-organisms
The heart (CGMCC)；The preservation time is：On September 24th, 2015.
Allotrophic nitrobacteria belongs to acinetobacter (Acinetobacter), is named as：Acinetobacter
sp.Y1；Preservation registration number is：CGMCC No.6563；Preservation mechanism is：China Committee for Culture Collection of Microorganisms is common
Microorganism center (CGMCC)；The preservation time is：On September 14th, 2012.
Wherein, the ratio between the bacterium solution volume of the pseudomonad Z1 and acinetobacter calcoaceticus Y1 is 1:1.
Specific implementation mode 2
Implement a kind of system of the composite bacteria agent of removal aniline and ammonia nitrogen while be used for described in above-mentioned specific implementation mode 1
Preparation Method, the preparation method follow these steps to carry out：
(1) preparation method of pseudomonad Z1
It is inoculated into 100mL fluid nutrient mediums from a small amount of thalline of picking on the inclined-plane of 4 DEG C of preservations, at 120rpm, 30 DEG C
Shaken cultivation 18h, until bacteria suspension OD600Stop culture when value is 0.6~0.8, obtains pseudomonad Z1 bacterium solutions；The vacation unit cell
The medium component of bacterium Z1 is：KH2PO41.0g/L；K2HPO41.0g/L；NaCl 1.0g/L；MgSO40.005g/L；
MnSO40.0017g/L；H3BO30.00116g/L；ZnSO4·7H2O 0.00115g/L；FeSO4·7H2O 0.003g/L；Aniline
0.4g/L；PH value is 7.0-7.2；
(2) preparation method of acinetobacter calcoaceticus Y1
It is inoculated into 100mL fluid nutrient mediums from a small amount of thalline of picking on the inclined-plane of 4 DEG C of preservations, at 120rpm, 30 DEG C
Shaken cultivation 14h, until bacteria suspension OD600Stop culture when value is 1.0~1.2, obtains acinetobacter calcoaceticus Y1 bacterium solutions；The not lever
The medium component of bacterium Y1 is：Sodium citrate 4.902g/L；(NH4)2SO40.472g/L；MgSO40.05g/L；K2HPO40.2g/
L；NaCl 0.12g/L；MnSO40.01g/L；FeSO4·7H2O 0.01g/L；PH value is 7.0-7.2；
(3) the pseudomonad Z1 and acinetobacter calcoaceticus Y1 prepared above-mentioned steps presses 1:1 volume ratio is mixed, and is obtained micro-
Biological compound fungi agent.
Specific implementation mode 3
The composite bacteria agent of removal aniline and ammonia nitrogen answers while implementing a kind of be used for described in above-mentioned specific implementation mode 1
With, or implement removal aniline and the preparation of ammonia nitrogen composite bacteria agent while a kind of be used for described in above-mentioned specific implementation mode 2
The application of composite bacteria agent prepared by method, the method for the application are the complex microbial inoculums that will prepare by volume 4%
Inoculum concentration, composite bacteria agent is inoculated in aniline waste water, for removing aniline and its aniline simultaneously and being discharged in degradation process
Ammonia nitrogen.
Wherein composite bacteria agent is inoculated in aniline waste water be by pseudomonad Z1 and acinetobacter calcoaceticus Y1 by volume be 1:1
It is inoculated in aniline waste water, composite bacteria agent while aniline degradation and ammonia nitrogen, after 48h, degradation rate of aniline is 93.54%, COD degradation
Rate is 88.41%, and ammonia nitrogen surplus is 4.39mg/L.
The specific implementation mode further illustrated the present invention below by specific embodiment.
Embodiment 1
A kind of composite bacteria agent provided by the present invention includes pseudomonad (Pseudomonas sp.) Z1 and acinetobacter calcoaceticus
(Acinetobacter sp.) Y1, pseudomonad Z1 and acinetobacter calcoaceticus Y1 is to be cultivated respectively by respective condition of culture,
Then it is 1 by volume:1 mixes and manufactured complex microbial inoculum.
(1) preparation method of pseudomonad Z1 bacterium solutions is as follows：
It is inoculated into 100mL fluid nutrient mediums from a small amount of thalline of picking on the inclined-plane of 4 DEG C of preservations, at 120rpm, 30 DEG C
Shaken cultivation 18h, until bacteria suspension OD600Stop culture when value is 0.6~0.8, obtains pseudomonad Z1 bacterium solutions；The vacation unit cell
The medium component of bacterium Z1 is：KH2PO41.0g/L；K2HPO41.0g/L；NaCl 1.0g/L；MgSO40.005g/L；
MnSO40.0017g/L；H3BO30.00116g/L；ZnSO4·7H2O 0.00115g/L；FeSO4·7H2O 0.003g/L；Benzene
Amine, 0.4g/L；PH value is 7.0-7.2；
(2) preparation method of acinetobacter calcoaceticus Y1 bacterium solutions is as follows：
It is inoculated into 100mL fluid nutrient mediums from a small amount of thalline of picking on the inclined-plane of 4 DEG C of preservations, at 120rpm, 30 DEG C
Shaken cultivation 14h, until bacteria suspension OD600Stop culture when value is 1.0~1.2, obtains acinetobacter calcoaceticus Y1 bacterium solutions；The not lever
The medium component of bacterium Y1 is：Sodium citrate 4.902g/L；(NH4)2SO40.472g/L；MgSO40.05g/L；K2HPO40.2g/
L；NaCl 0.12g/L；MnSO40.01g/L；FeSO4·7H2O 0.01g/L；PH value is 7.0-7.2；
Embodiment 2
Application of the composite bacteria agent of the present invention to the degradation of aniline, ammonia nitrogen and COD
Composite bacteria agent is inoculated in the fluid nutrient medium of simulation aniline waste water by the volume ratio of inoculum concentration 4%, as reality
Test group；Z1 is inoculated in same simulation aniline waste water by identical inoculative proportion, as a contrast.30 DEG C, under the conditions of 120rpm
Shaken cultivation.Timing sampling detects aniline, ammonia nitrogen and COD.As a result attached drawing 1, attached drawing 2 and attached drawing 3 are seen respectively.
From attached drawing 1 as can be seen that compared with Z1 aniline degradations, composite bacteria agent slightly influences the degradation rate of aniline, but
Degradation rate is had little effect.Figure it is seen that Z1 can accumulate a certain amount of ammonia nitrogen during aniline degradation, but
Mix bacterium agent does not accumulate ammonia nitrogen substantially during aniline degradation, illustrates that composite bacteria agent can remove aniline and aniline simultaneously
The ammonia nitrogen discharged in degradation process.From figure 3, it can be seen that compared with Z1, composite bacteria agent is fine to the degradation effect of COD.It is comprehensive
On, composite bacteria agent can remove the ammonia nitrogen that aniline and aniline discharge in degradation process simultaneously, and the benzene that can degrade well
The COD of amine waste water.
Wherein, the fluid nutrient medium of aniline waste water is simulated：Aniline 0.6g/L；Sodium citrate 3.16g/L；KH2PO41.0g/L；
K2HPO41.0g/L；NaCl 1.0g/L；MgSO40.005g/L；MnSO40.0017g/L；H3BO30.00116g/L；ZnSO4·
7H2O 0.00115g/L；FeSO4·7H2O 0.003g/L；PH value is 7.0-7.2.
Claims (6)
1. composite bacteria agent that is a kind of while removing aniline and ammonia nitrogen, the composite bacteria agent are thin by microphenomenon of degradation of aniline and heterotrophic nitrification
Bacterium is compound and constitutes, wherein：
Microphenomenon of degradation of aniline belongs to pseudomonas Pseudomonas；It is named as Pseudomonas sp.Z1；Preservation registration number
For：CGMCC No.11444；Preservation mechanism is：China Committee for Culture Collection of Microorganisms's common micro-organisms center；Preservation
Time is：On September 24th, 2015；
Allotrophic nitrobacteria belongs to acinetobacter Acinetobacter；It is named as Acinetobacter sp.Y1；Preservation
Registration number is：CGMCC No.6563；Preservation mechanism is：China Committee for Culture Collection of Microorganisms's common micro-organisms center；
The preservation time is：On September 14th, 2012.
2. the ratio between the bacterium solution volume of composite bacteria agent according to claim 1, the pseudomonad Z1 and acinetobacter calcoaceticus Y1 are
1:1。
3. a kind of preparation method of composite bacteria agent as claimed in claim 1 or 2, the preparation method is to follow these steps to carry out
's：
(1) preparation method of pseudomonad Z1
It is inoculated into 100mL fluid nutrient mediums from a small amount of thalline of picking on the inclined-plane of 4 DEG C of preservations, is vibrated at 120rpm, 30 DEG C
18h is cultivated, until stopping culture when bacteria suspension OD600 values are 0.6~0.8, obtains pseudomonad Z1 bacterium solutions；The pseudomonad
The medium component of Z1 is：KH2PO41.0g/L；K2HPO41.0g/L；NaCl1.0g/L；MgSO40.005g/L；
MnSO40.0017g/L；H3BO30.00116g/L；ZnSO4·7H2O0.00115g/L；FeSO4·7H2O0.003g/L；Aniline
0.4g/L；PH value is 7.0-7.2；
(2) preparation method of acinetobacter calcoaceticus Y1
It is inoculated into 100mL fluid nutrient mediums from a small amount of thalline of picking on the inclined-plane of 4 DEG C of preservations, is vibrated at 120rpm, 30 DEG C
14h is cultivated, until stopping culture when bacteria suspension OD600 values are 1.0~1.2, obtains acinetobacter calcoaceticus Y1 bacterium solutions；The acinetobacter calcoaceticus Y1
Medium component be：Sodium citrate 4.902g/L；(NH4)2SO40.472g/L；MgSO40.05g/L；K2HPO40.2g/L；
NaCl0.12g/L；MnSO40.01g/L；FeSO4·7H2O0.01g/L；PH value is 7.0-7.2；
(3) the pseudomonad Z1 and acinetobacter calcoaceticus Y1 prepared above-mentioned steps presses 1:1 volume ratio is mixed, and microorganism is obtained
Composite bacteria agent.
4. a kind of application for composite bacteria agent described in claim 1, the application is that the microorganism that will prepare is compound
Microbial inoculum is used for while removing the ammonia nitrogen that aniline and its aniline discharge in degradation process.
It is described while removing aniline and its aniline in degradation process 5. the application of composite bacteria agent according to claim 4
The ammonia nitrogen of release be by volume be 4% inoculum concentration, composite bacteria agent is inoculated in aniline waste water.
6. the application of composite bacteria agent according to claim 5, it is described composite bacteria agent is inoculated in aniline waste water be will be false
Monad Z1 and acinetobacter calcoaceticus Y1 is 1 by volume:1 is inoculated in aniline waste water.
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| CN101165167A (en) * | 2006-10-18 | 2008-04-23 | 中国科学院沈阳应用生态研究所 | Microbial inoculum for repairing amino benzenes polluted water and its preparation method and using method |
| CN101514330A (en) * | 2008-12-18 | 2009-08-26 | 浙江工业大学 | Achromobacter capable of degrading aniline and application thereof |
| CN102167448A (en) * | 2010-12-03 | 2011-08-31 | 中国地质大学（武汉） | Application of aniline degradation strain NO.3 |
| CN104528956A (en) * | 2015-01-22 | 2015-04-22 | 马金晔 | Composite microbial inoculant for treating coal chemical industry sewage and application thereof |
| CN104628225A (en) * | 2015-01-09 | 2015-05-20 | 张玉兰 | Treatment method of ammonia-nitrogen-containing industrial wastewater |
-
2015
- 2015-10-20 CN CN201510681384.2A patent/CN105199992B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101165167A (en) * | 2006-10-18 | 2008-04-23 | 中国科学院沈阳应用生态研究所 | Microbial inoculum for repairing amino benzenes polluted water and its preparation method and using method |
| CN101514330A (en) * | 2008-12-18 | 2009-08-26 | 浙江工业大学 | Achromobacter capable of degrading aniline and application thereof |
| CN102167448A (en) * | 2010-12-03 | 2011-08-31 | 中国地质大学（武汉） | Application of aniline degradation strain NO.3 |
| CN104628225A (en) * | 2015-01-09 | 2015-05-20 | 张玉兰 | Treatment method of ammonia-nitrogen-containing industrial wastewater |
| CN104528956A (en) * | 2015-01-22 | 2015-04-22 | 马金晔 | Composite microbial inoculant for treating coal chemical industry sewage and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 异养硝化-好氧反硝化菌Acinetobacter sp.Y1的筛选和脱氮性能的研究;宋宇杰;《中国优秀硕士学位论文全文数据库工程科技I辑（电子期刊）》;20140815(第08期);第37页 * |
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