CN107164276A - One plant tolerance zinc ion toxicity pseudomonas aeruginosa and its application - Google Patents
One plant tolerance zinc ion toxicity pseudomonas aeruginosa and its application Download PDFInfo
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- CN107164276A CN107164276A CN201710475189.3A CN201710475189A CN107164276A CN 107164276 A CN107164276 A CN 107164276A CN 201710475189 A CN201710475189 A CN 201710475189A CN 107164276 A CN107164276 A CN 107164276A
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- C12R2001/38—Pseudomonas
- C12R2001/385—Pseudomonas aeruginosa
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/02—Aerobic processes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
- C02F2101/163—Nitrates
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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
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The present invention relates to one plant of pseudomonas aeruginosa (Pseudomonas aeruginosa) for being resistant to zinc ion toxicity and its application.The invention provides the pseudomonas aeruginosa (Pseudomonas aeruginosa) that one plant has zinc ion tolerance, its preserving number is CGMCC No.14063.When zinc ion concentration is in the range of 0~100mg/L in sewage, the bacterial strain can be using nitrate nitrogen as only nitrogen source, under aerobic condition, and to nitrate nitrogen clearance up to 100%, the clearance of total nitrogen is also up to 100%.The present invention has simple operation, and the advantages of nitric efficiency is high, the influence to optimization sewage biological treatment system and decrease zinc ion to sewage denitrification system is significant.
Description
Technical field
The invention belongs to field of biological sewage treatment, and in particular to one kind is resistant to zinc ion toxicity under aerobic condition
And can be by the pseudomonas aeruginosa of the nitrate nitrogen simultaneous removing in water body and its application in sewage disposal.
Background technology
Industrial to continue to develop with Chinese Urbanization, industrialized acceleration, substantial amounts of industrial wastewater is produced, right
Human lives and health generate great threat.Because the Industrial Wastewater Treatment ability of China can not still keep up with industrial water
Scale, cause still there is part industrial wastewater is unprocessed to be directly discharged into town sewage pipe network.Zinc ion is important as one kind
Industrial materials, be widely used in chemical industry, metallurgy, plating, process hides and coating industry, these industries can produce substantial amounts of contain
Zinc waste water, these waste water also inevitably enter urban wastewater treatment firm.Research shows that Zn (II) concentration is more than 10mg/L
N in acting on conventional denitrification2O reduction process, denitrification rate and denitrifying microorganism activity produces obvious suppression
System, meanwhile, change biological community structure, and then influence the effluent characteristics of town sewage treatment system.
In recent years, aerobic denitrification (process that nitrate nitrogen is reduced to nitrogenous gas under aerobic conditions)
Be found to be realized in single aerobic reactor synchronous nitration and denitrification provide theoretical foundation.Since first plant of aerobic anti-nitre
After change bacterium T.pantotropha is successfully separated, more high-efficiency aerobic denitrification bacterial strains are separated in recent years, and are answered
For in actual sewage processing system.Aerobic denitrification as a kind of novel biological dinitrification technology, because its technique is simple,
Denitrification effect is good, is not required to add the advantages such as soda acid and is developed rapidly in recent years.
Enter sanitary sewage for zinc-containing water, how the problem of Zn (II) and nitrate nitrogen coexist is effectively prevented from zinc
The influence that ion pair nitrate nitrogen is removed turns into focus of concern, therefore, excavates with the aerobic of tolerance zinc ion toxicity
Denitrifying bacteria, and be applied in sewage disposal, it will zinc ion is effectively cut down to the potential of bio-denitrifying sewage process
Risk, to ensure that the normally operation of town sewage treatment system is significant.
The content of the invention
It is strong by the bacterial strain it is an object of the invention to provide a kind of high-efficiency aerobic denitrogenation bacterial strain for being resistant to zinc ion toxicity
Change the Nitrogen removal effect of sewage disposal system, to cut down the presence of Treatment with High Concentration Zinc ion to sewage disposal biological denitrification process
Influence.
Pseudomonas aeruginosa (Pseudomonas aeruginosa) provided by the present invention, which is one plant, to be had single good
The aerobic denitrifying bacteria of nitrate nitrogen removal ability is realized in oxygen environment.
The technical scheme is that：
The pseudomonas aeruginosa (Pseudomonas aeruginosa) with zinc ion tolerance that the present invention is provided,
China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC) is preserved on April 24th, 2017,
Address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, and its preserving number is CGMCC
No.14063。
Pseudomonas aeruginosa (Pseudomonas aeruginosa) as described above, it is characterised in that：Different Zinc from
Under the conditions of sub- concentration level, strain Pseudomonas aeruginosa can still keep complete eucaryotic cell structure, cell membrane
It is not destroyed.
Pseudomonas aeruginosa (Pseudomonas aeruginosa) as described above, it is characterised in that：The bacterial strain energy
Enough using nitrate nitrogen as nitrogen source under Treatment with High Concentration Zinc ionic conditions, aerobic denitrification is carried out by carbon source of organic matter, so that
Nitrate nitrogen is removed.
Application of the pseudomonas aeruginosa (Pseudomonas aeruginosa) as described above in sewage disposal, its
It is characterised by：Pseudomonas aeruginosa (Pseudomonas aeruginosa) is added in sewage containing zinc, and is added in right amount
Carbon source is aerated, you can realize the removal of nitrate from waste water nitrogen.
The process described above, it is characterised in that：It is 30 DEG C to control the temperature in the sewage containing zinc, and pH value is 7.5,
Dissolved oxygen is 6mg/L, and initial C/N ratios are 4.
The process described above, it is characterised in that：When zinc ion concentration scope is 0~100mg/L in sewage, nitrate
The clearance of nitrogen is up to 100%, and the clearance of total nitrogen is also up to 100%.
The beneficial effects of the invention are as follows：
(1) pseudomonas aeruginosa (Pseudomonas aeruginosa) of the invention can be in Treatment with High Concentration Zinc ionic conditions
Under, using nitrate nitrogen as nitrogen source, aerobic denitrification is carried out by carbon source of organic matter, the efficient removal of nitrate nitrogen is realized；
Solve the problem of biological denitrificaion needs to take anoxic denitrification, aerobic nitrification segment processing in conventional waste water processing；In addition,
Technological process is simplified, the cost of equipment and investment is saved, therefore, with huge economic benefit and environmental benefit；
(2) by the biological treatment of the inoculation to the sewage containing zinc, under 0~150mg/L Zn (II) existence condition,
It is preferred that, under 0~100mg/L Zn (II) existence condition, the clearance of nitrate nitrogen illustrates that bacterial strain has up to 100%
Stronger tolerance zinc ion toxicity and aerobic denitrifying capacity.This characteristic causes the bacterial strain in town sewage treatment system
Practicality is greatly enhanced.
Brief description of the drawings
Fig. 1 is scanning electron microscope image of the pseudomonas aeruginosa under zinc ion existence condition.
Fig. 2 is aerobic denitrification characteristic of the pseudomonas aeruginosa under 10mg/L Zn (II) existence condition.
Fig. 3 is aerobic denitrification characteristic of the pseudomonas aeruginosa under 50mg/L Zn (II) existence condition.
Fig. 4 is aerobic denitrification characteristic of the pseudomonas aeruginosa under 100mg/L Zn (II) existence condition.
Embodiment
With reference to specific embodiment, the invention will be further described, but the present invention is not limited to following examples.
It is conventional method unless otherwise specified in following embodiments.
In following embodiments, the percentage composition is weight/mass percentage composition unless otherwise specified.
Mode of appearance of the embodiment 1. using scanning electron microscopic observation pseudomonas aeruginosa in the presence of zinc ion
High-efficiency aerobic denitrification bacterial strain used in this experiment is pseudomonas aeruginosa (Pseudomonas aeruginosa), its
It is isolated from percolate from garbage filling field processing system.
Pseudomonas aeruginosa (Pseudomonas aeruginosa) is inoculated in 1L LB culture mediums to (every liter contains NaCl
5g, tryptone 10g, yeast extract 5g), prevent the intrusion of miscellaneous bacteria and keep the growth vigor of thalline, carry out enrichment training
Support.Obtained bacterium solution centrifugation will be cultivated, washed with 0.5% NaCl three times, optical density OD is made600For 1-2 bacteria suspension.So
Bacteria suspension is inoculated in containing in 0,10,50,100mg/L Zn (II) LB culture mediums respectively by 10% inoculum concentration again afterwards,
Sampled after 8h and 5min removal supernatants are centrifuged under the conditions of 8000rpm and 4 DEG C, and two are cleaned with PBS (phosphate buffer)
It is secondary.It is dispersed in after fixing 1h, ethanol gradient elution with 2.5% glutaraldehyde in absolute ethyl alcohol, takes a small amount of drop on silicon chip to dry,
Cellular morphology is observed under ESEM (SEM).
As a result as shown in figure 1, when zinc ion concentration scope be 0~100mg/L when, strain Pseudomonas
Complete and smooth surface is presented in aeruginosa, illustrates that strain cell form has stronger tolerance to zinc ion.
Aerobic denitrification characteristic of the pseudomonas aeruginosa of embodiment 2. under 10mg/L Zn (II) existence condition
Denitrification capability test media (BM) formula is in every liter of water：8.45g CH3COONa,0.63g NH4Cl,
0.61g NaNO3,1.76g K2HPO4·3H2O,0.20g MgSO4·7H2O,0.02g CaCl2,0.005g FeSO4·7H2O,
0.1mL trace element solutions.Medium's PH Value is adjusted to 7.5, sterilize 30min at 121 DEG C.
Pseudomonas aeruginosa (Pseudomonas aeruginosa) is inoculated in the BM cultures containing 10mg/L Zn (II)
In base, concussion and cultivate is carried out under the conditions of 30 DEG C and 150rpm, 100 μ L static headspace gas are extracted respectively every 5h apparatus valve injection pins
Body is used to determine N2O, being extracted 2mL gases with asepsis injector and be injected into the pure helium bags of 2L is used to determine NO.At the same time,
2mL bacteria suspensions are extracted with asepsis injector, bacterium solution 8000rpm at 4 DEG C centrifuges 5min, takes supernatant to be used to analyze ammonia nitrogen, nitre
Hydrochlorate nitrogen, nitrite nitrogen concentration.
As a result as shown in Fig. 2 100mg/L nitrate nitrogens are utilized by bacterial strain immediately after inoculation, and it is complete in 10h
Consumption, the average removal rate of nitrate nitrogen is 10.0mg/L/h.With the reduction of nitrate nitrogen, nitrite nitrogen is quickly accumulated
It is tired, and peak 29.5mg/L is reached in 5h, then it is reduced in 10h.N2O is also gradually accumulated, and peak is reached in 10h
80.2mg/L, then in 35h by Restore All.It must be noted that the peak of the intermediate product NO accumulations of denitrification process is
1.2 μ g/L, only account for the 0.0012% of nitrate nitrogen removal amount, illustrate that bacterial strain has under the conditions of 10mg/L Zn (II) efficiently good
The denitrifying ability of oxygen.
Aerobic denitrification characteristic of the pseudomonas aeruginosa of embodiment 3. under 50mg/L Zn (II) existence condition
Pseudomonas aeruginosa (Pseudomonas aeruginosa) is inoculated in the BM cultures containing 50mg/L Zn (II)
In base, its aerobic denitrification capability is tested.As a result as shown in figure 3,100mg/L nitrate nitrogens are after inoculation immediately by bacterial strain profit
With, and be totally consumed in 15h, the average removal rate of nitrate nitrogen is 6.67mg/L/h.With going back for nitrate nitrogen
Original, nitrite nitrogen is gradually accumulated, and reaches peak 35.5mg/L in 10h, is then reduced in 20h.N2O is also gradually accumulated
It is tired, peak 84.4mg/L is reached in 20h, then in 45h by Restore All.It must be noted that the now centre of denitrification process
The peak of product NO accumulations is 3.2 μ g/L, only accounts for the 0.0032% of nitrate nitrogen removal amount.It can thus be seen that bacterial strain
Still there is the denitrifying ability of high-efficiency aerobic under the conditions of 50mg/L Zn (II).
Aerobic denitrification characteristic of the pseudomonas aeruginosa of embodiment 4. under 100mg/L Zn (II) existence condition
Pseudomonas aeruginosa (Pseudomonas aeruginosa) is inoculated in the BM trainings containing 100mg/L Zn (II)
Support in base, test its aerobic denitrification capability.As a result as shown in figure 4,100mg/L nitrate nitrogens are after inoculation immediately by bacterial strain
Utilize, and be totally consumed in 20h, the average removal rate of nitrate nitrogen is 5.0mg/L/h.With going back for nitrate nitrogen
Original, nitrite nitrogen is gradually accumulated, and reaches peak 40.2mg/L in 15h, is then reduced in 30h.Meanwhile, N2O also by
Gradually accumulate, peak 89.5mg/L is reached in 25h, then in 55h by Restore All.It must be noted that now denitrification process
The peak of intermediate product NO accumulations is 6.3 μ g/L, only accounts for the 0.0063% of nitrate nitrogen removal amount.It can thus be seen that
Bacterial strain still has the denitrifying ability of high-efficiency aerobic under the conditions of 100mg/L Zn (II).
Aerobic denitrification characteristic of the pseudomonas aeruginosa of embodiment 5. under 150mg/L Zn (II) existence condition
Pseudomonas aeruginosa (Pseudomonas aeruginosa) is inoculated in the BM trainings containing 150mg/L Zn (II)
Support in base, test its aerobic denitrification capability.The growth of the bacterial strain is significantly suppressed, and nitrate nitrogen is hardly gone
Remove, it can thus be seen that 150mg/L Zn (II) are the upper concentrations of the bacterial strain, bacterial strain does not possess aerobic anti-under this condition
The ability of nitrification.But as can be seen that by the biological treatment of the inoculation to the sewage containing zinc from above-described embodiment 1-4,
Under 0~100mg/L Zn (II) existence condition, the clearance of nitrate nitrogen illustrates that bacterial strain has stronger resistance to up to 100%
By zinc ion toxicity and aerobic denitrifying capacity (being shown in Table 1).
Nitrogen removal characteristics of the pseudomonas aeruginosa of table 1 under various concentrations Zn (II) existence condition
| Zn(II)(mg/L) | 30h nitrate nitrogens clearance (%) | 30h nitrogen removal rates (%) |
| 10 | 100% | 100% |
| 50 | 100% | 100% |
| 100 | 100% | 100% |
Application of the pseudomonas aeruginosa of embodiment 6. in zinc-containing water
Pseudomonas aeruginosa is added into activated Sludge System Treatment of Zinc-containing Wastewater, continuous aeration holding system dissolved oxygen is
6 mg/L, waste water quality is as follows：PH is 7.5, nitrate nitrogen content 100mg/L, Zn (II) 100mg/L.As a result as shown in table 2,
In ordinary activated sludge system, denitrification effect receives large effect under Zn (II) existence condition.However, through verdigris
System after pseudomonad reinforcing, 24h nitrate nitrogen clearance is 70.7%, and nitrogen removal rate is 63.4%；36h nitre
Hydrochlorate nitrogen removal efficiency is 94.8%, and nitrogen removal rate is 92.4%.
The pseudomonas aeruginosa of table 2. is applied to the biological denitrificaion result in activated sludge system
As can be seen here, sanitary sewage, the problem of Zn (II) and nitrate nitrogen coexist, institute of the present invention are entered for zinc-containing water
The bacterial strain being related to can effectively avoid the influence that zinc ion is removed to nitrate nitrogen, it is possible to be widely used in sewage
In processing, potential risk of the zinc ion to bio-denitrifying sewage process can be effectively cut down, to ensure town sewage treatment system
Normally run significant.
It should be noted that above example is intended merely to, the present invention is described in further detail, is not intended to pair
What the present invention was defined, in the design for not departing from the present invention and the scope of spirit, those of ordinary skill in the art can be with
Various improvement or change are carried out, the protection domain of appended claims of the invention is still fallen within.
Claims (9)
1. the application of the pseudomonas aeruginosa of one plant of tolerance zinc ion toxicity, it is characterised in that：It is thin by one plant of aerobic denitrification
The Nitrogen removal effect of bacterium pseudomonas aeruginosa (Pseudomonas aeruginosa) enhanced sewage processing system, can cut down
Measure influence of the presence of zinc ion to sewage disposal biological denitrification process.
2. the application of pseudomonas aeruginosa (Pseudomonas aeruginosa) as claimed in claim 1, it is characterised in that：
The preserving number of the bacterial strain is CGMCC No.14063, has preferable tolerance to the zinc ion of various concentrations level.
3. the application of pseudomonas aeruginosa (Pseudomonas aeruginosa) according to claim 1, its feature exists
In：The bacterial strain can be carried out aerobic anti-under different zinc ion concentration levels using nitrate nitrogen as nitrogen source by carbon source of organic matter
Nitrification, so that nitrate nitrogen be removed.
4. pseudomonas aeruginosa (Pseudomonas aeruginosa) as claimed in claim 1 is in bio-denitrifying sewage
Using, it is characterised in that：Pseudomonas aeruginosa (Pseudomonas aeruginosa) is added in sewage containing zinc, and is added
Proper amount of carbon source is aerated, you can realize the removal of nitrate from waste water nitrogen.
5. application according to claim 4, it is characterised in that：It is 30 DEG C to control the temperature in the sewage containing zinc.
6. application according to claim 4, it is characterised in that：It is 7.5 to control the pH value in the sewage containing zinc.
7. application according to claim 4, it is characterised in that：It is 6mg/L to control the dissolved oxygen in the sewage containing zinc.
8. application according to claim 4, it is characterised in that：It is 4 to control the initial C/N ratios in the sewage containing zinc.
9. application according to claim 4, it is characterised in that：Zinc ion concentration scope contained by sewage is 0~100mg/
L, the clearance of nitrate nitrogen is up to 100% under aerobic environment, and the clearance of total nitrogen is also up to 100%.
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Cited By (4)
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| CN109055252A (en) * | 2017-11-13 | 2018-12-21 | 鼎正新兴生物技术（天津）有限公司 | Heterotrophic nitrification-aerobic denitrification composite microbial preparation and preparation method thereof |
| CN114107136A (en) * | 2021-12-06 | 2022-03-01 | 华北电力大学 | Pseudomonas stutzeri with aerobic denitrification and greenhouse gas emission reduction functions |
| CN114591853A (en) * | 2022-02-24 | 2022-06-07 | 安徽师范大学 | Heavy metal-resistant aerobic denitrification strain with bisphenol A and nitrogen removal functions and application thereof |
| CN115322936A (en) * | 2022-09-21 | 2022-11-11 | 北京大学 | Aerobic denitrifying bacterium with sulfonamide antibiotic degradation capability and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109055252A (en) * | 2017-11-13 | 2018-12-21 | 鼎正新兴生物技术（天津）有限公司 | Heterotrophic nitrification-aerobic denitrification composite microbial preparation and preparation method thereof |
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| CN114591853A (en) * | 2022-02-24 | 2022-06-07 | 安徽师范大学 | Heavy metal-resistant aerobic denitrification strain with bisphenol A and nitrogen removal functions and application thereof |
| CN115322936A (en) * | 2022-09-21 | 2022-11-11 | 北京大学 | Aerobic denitrifying bacterium with sulfonamide antibiotic degradation capability and application thereof |
| CN115322936B (en) * | 2022-09-21 | 2023-10-27 | 北京大学 | Aerobic denitrifying bacterium with sulfonamide antibiotic degradation capability and application thereof |
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