CN110283744A - One plant of achromobacter for being resistant to zinc ion toxicity and its application - Google Patents
One plant of achromobacter for being resistant to zinc ion toxicity and its application Download PDFInfo
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- CN110283744A CN110283744A CN201910552280.XA CN201910552280A CN110283744A CN 110283744 A CN110283744 A CN 110283744A CN 201910552280 A CN201910552280 A CN 201910552280A CN 110283744 A CN110283744 A CN 110283744A
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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
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/025—Achromobacter
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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
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Abstract
The achromobacter (Achromobacter sp.) for being resistant to zinc ion toxicity the present invention relates to one plant and its application.The present invention provides one plant of achromobacter (Achromobacter sp.) with zinc ion tolerance, deposit number is CGMCC No.2964.When zinc ion concentration is within the scope of 0~50mg/L in sewage, which can be using nitrate nitrogen as only nitrogen source, under aerobic condition, and to nitrate nitrogen removal rate up to 100%, the removal rate of total nitrogen is also up to 100%.The present invention has many advantages, such as that operation is convenient, and nitric efficiency is high, and the influence to optimization sewage biological treatment system and decrease zinc ion to sewage denitrification system is of great significance.
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 it can be by the achromobacter of the nitrate nitrogen simultaneous removing in water body and its application in sewage treatment.
Background technique
With Chinese Urbanization, industrialized acceleration, industrial continuous development, a large amount of industrial wastewater is generated, right
Human lives and health produce great threat.Since the Industrial Wastewater Treatment ability in China cannot still keep up with industrial water
Scale leads to still have part industrial wastewater is unprocessed to be directly discharged into town sewage pipe network.Zinc ion is as a kind of important
Industrial materials are widely used in chemical industry, metallurgy, plating, process hides and coating industry, these industries can generate largely useless containing zinc
Water, these waste water also inevitably enter urban wastewater treatment firm.Studies have shown that Zn (II) concentration be more than 10mg/L can be to biography
N in system denitrification2Reduction process, denitrification rate and the denitrifying microorganism activity of O generates apparent inhibition, meanwhile,
Change biological community structure, and then influences the effluent characteristics of town sewage treatment system.
In recent years, aerobic denitrification (nitrate nitrogen is reduced to under aerobic conditions the process of nitrogenous gas)
It is found to be in single aerobic reactor and realizes that synchronous nitration and denitrification provides theoretical basis.Since first plant of aerobic denitrification
After bacterium T.pantotropha is successfully separated, more high-efficiency aerobic denitrification bacterial strains are separated in recent years, and are applied to
In actual sewage processing system.Aerobic denitrification is as a kind of new biological nitrogen removal technology, because its simple process, denitrogenation are imitated
Fruit is good, is not required to add the advantages such as soda acid and is developed rapidly in recent years.
For the problem that zinc-containing water enters sanitary sewage, Zn (II) and nitrate nitrogen are coexisted, and how to be effectively prevented from zinc
The influence of ion pair nitrate nitrogen removal becomes focus concerned by people, and therefore, excavating has the aerobic of tolerance zinc ion toxicity
Denitrifying bacteria, and be applied in sewage treatment, it will zinc ion is effectively cut down to the potential of bio-denitrifying sewage process
Risk, to ensure that the normal operation of town sewage treatment system is of great significance.
Summary of the invention
It is strong by the bacterial strain the purpose of the present invention is 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 treatment biological denitrification process
It influences.
Achromobacter (Achromobacter sp.) provided by the present invention, which is one plant, to be had in single aerobic environment in fact
The aerobic denitrifying bacteria of existing nitrate nitrogen removal ability.
The technical scheme is that:
Achromobacter (Achromobacter sp.) provided by the invention with zinc ion tolerance, deposit number
For CGMCC No.2964.
Achromobacter (Achromobacter sp.) as described above, it is characterised in that: horizontal in different zinc ion concentrations
Under the conditions of, strains A chromobacter sp. is still able to maintain complete eucaryotic cell structure, and cell membrane is not destroyed.
Achromobacter (Achromobacter sp.) as described above, it is characterised in that: the bacterial strain can be in high concentration
Using nitrate nitrogen as nitrogen source under the conditions of zinc ion, aerobic denitrification is carried out by carbon source of organic matter, thus by nitrate nitrogen
Removal.
Application of the achromobacter (Achromobacter sp.) in sewage treatment as described above, it is characterised in that:
Achromobacter (Achromobacter sp.) is added in sewage containing zinc, and adds suitable carbon source and is aerated, Ji Keshi
The removal of existing nitrate from waste water nitrogen.
The process described above, it is characterised in that: controlling the temperature in the sewage containing zinc is 30 DEG C, and pH value 7.5 is molten
Solution oxygen is 6mg/L, and initial C/N ratio is 4.
The process described above, it is characterised in that: when zinc ion concentration range is 0~50mg/L in sewage, nitrate nitrogen
Removal rate up to 100%, the removal rate of total nitrogen is also up to 100%.
The beneficial effects of the present invention are:
(1) achromobacter of the invention (Achromobacter sp.) can be under Treatment with High Concentration Zinc ionic conditions, with nitric acid
Salt nitrogen is nitrogen source, carries out aerobic denitrification by carbon source of organic matter, realizes the efficient removal of nitrate nitrogen；Solves tradition
Biological denitrificaion needs the problem of taking anoxic denitrification, aerobic nitrification segment processing in wastewater treatment；In addition, simplifying technique stream
Journey saves the cost of equipment and investment, therefore, has huge economic benefit and environmental benefit；
(2) excellent under 0~50mg/L Zn (II) existence condition by the biological treatment of the strain inoculated to the sewage containing zinc
Choosing, under 0~25mg/L Zn (II) existence condition, the removal rate of nitrate nitrogen illustrates that bacterial strain has up to 100% in 25h
Stronger tolerance zinc ion toxicity and aerobic denitrifying capacity.This characteristic makes the bacterial strain in the reality of town sewage treatment system
It is greatly enhanced with property.
Detailed description of the invention
Fig. 1 is scanning electron microscope image of the achromobacter under zinc ion existence condition.
Fig. 2 is aerobic denitrification characteristic of the achromobacter under 10mg/L Zn (II) existence condition.
Fig. 3 is aerobic denitrification characteristic of the achromobacter under 25mg/L Zn (II) existence condition.
Fig. 4 is aerobic denitrification characteristic of the achromobacter under 50mg/L Zn (II) existence condition.
Specific embodiment
The present invention is further explained in the light of specific embodiments, but the present invention is not limited to following embodiments.
It is conventional method unless otherwise specified in following embodiments.
In following embodiments, the percentage composition is mass percentage unless otherwise specified.
Mode of appearance of the embodiment 1. using scanning electron microscopic observation achromobacter in the presence of zinc ion
High-efficiency aerobic denitrification bacterial strain used in this experiment is achromobacter (Achromobacter sp.), is isolated from rubbish
Rubbish landfill percolate processing system.
Achromobacter (Achromobacter sp.) is inoculated in the LB culture medium of 1L (every liter of 5g containing NaCl, pancreas egg
White peptone 10g, yeast extract 5g), it prevents the intrusion of miscellaneous bacteria and keeps the growth vigor of thallus, carry out enrichment culture.It will culture
Obtained bacterium solution centrifugation, is washed three times with 0.5% NaCl, optical density OD is made600For the bacteria suspension of 1-2.Then bacterium is hanged again
Liquid is inoculated in the LB culture medium containing 25mg/L Zn (II) respectively by 10% inoculum concentration, and sampling is in 8000rpm and 4 after 8h
It is centrifuged 5min under the conditions of DEG C and removes supernatant, and is cleaned twice with PBS (phosphate buffer).1h is fixed with 2.5% glutaraldehyde,
It is dispersed in dehydrated alcohol after ethanol gradient elution, takes a small amount of drop to dry on silicon wafer, observe cell at scanning electron microscope (SEM)
Form.
As a result as shown in Figure 1, when zinc ion concentration be 25mg/L when, strains A chromobacter sp. present completely and
Smooth surface illustrates that strain cell form has stronger tolerance to zinc ion.
Aerobic denitrification characteristic of 2. achromobacter of embodiment 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 CaCl2,0.005gFeSO4·7H2O,
0.1mL trace element solution.Medium's PH Value is adjusted to 7.5, in 121 DEG C of sterilizing 30min.
Achromobacter (Achromobacter sp.) is inoculated in the BM culture medium containing 10mg/L Zn (II), 30
DEG C and 150rpm under the conditions of carry out shake culture, extract 100 μ L headspace gas respectively for measuring every 5h apparatus valve injection needle
N2O extracts 2mL gas with asepsis injector and is injected into the pure helium bag of 2L for measuring NO.At the same time, asepsis injector is used
2mL bacteria suspension is extracted, bacterium solution 8000rpm at 4 DEG C is centrifuged 5min, takes supernatant for analyzing ammonia nitrogen, nitrate nitrogen, nitrous acid
Salt nitrogen concentration.
As a result as shown in Fig. 2, 100mg/L nitrate nitrogen is utilized by bacterial strain immediately after inoculation, and disappeared completely in 10h
Consumption, the average removal rate of nitrate nitrogen are 10.0mg/L/h.With the reduction of nitrate nitrogen, nitrite nitrogen Rapid Accumulation,
And reach peak 32.1mg/L in 10h, then it is reduced in 20h.N2O is also gradually accumulated, and reaches peak in 10h
Then 0.42mg/L is all restored in 25h.The peak of the intermediate product NO accumulation of denitrification process is 82.5 μ g/L,
Accumulation is less, illustrates that bacterial strain remains to preferably play aerobic denitrifying ability under the conditions of 10mg/L Zn (II) is existing.
Aerobic denitrification characteristic of 3. achromobacter of embodiment under 25mg/L Zn (II) existence condition
Achromobacter (Achromobacter sp.) is inoculated in the BM culture medium containing 25mg/L Zn (II), is tested
Its aerobic denitrification capability.As a result as shown in figure 3,100mg/L nitrate nitrogen is utilized by bacterial strain immediately after inoculation, and in 25h
It is inside totally consumed, the average removal rate of nitrate nitrogen is 4mg/L/h.With the reduction of nitrate nitrogen, nitrite nitrogen by
It gradually accumulates, and reaches peak 33.4mg/L in 15h, be then reduced in 25h.N2O is also gradually accumulated, and reaches highest in 20h
Then value 1.01mg/L is all restored in 45h.It must be noted that the intermediate product NO accumulation of denitrification process is most at this time
High level is 83.1 μ g/L, only accounts for the 0.083% of nitrate nitrogen removal amount.It can thus be seen that bacterial strain is in 25mg/L Zn (II) item
Still there is the denitrifying ability of high-efficiency aerobic under part.
Aerobic denitrification characteristic of 4. achromobacter of embodiment under 50mg/L Zn (II) existence condition
Achromobacter (Achromobacter sp.) is inoculated in the BM culture medium containing 50mg/L Zn (II), is tested
Its aerobic denitrification capability.As a result as shown in figure 4,100mg/L nitrate nitrogen is utilized by bacterial strain immediately after inoculation, and in 25h
It is inside totally consumed, the average removal rate of nitrate nitrogen is 4.0mg/L/h.With the reduction of nitrate nitrogen, nitrite nitrogen
It gradually accumulates, and reaches peak 33.5mg/L in 20h, be then reduced in 30h.Meanwhile N2O is also gradually accumulated, and is reached in 25h
To peak 1.23mg/L, then all restored in 35h.It must be noted that the intermediate product NO accumulation of denitrification process at this time
The peak of amount is 17.3 μ g/L, only accounts for the 0.017% of nitrate nitrogen removal amount.It can thus be seen that bacterial strain is in 50mg/L Zn
(II) still there is the denitrifying ability of high-efficiency aerobic under the conditions of.
Aerobic denitrification characteristic of 5. achromobacter of embodiment under 100mg/L Zn (II) existence condition
Achromobacter (Achromobacter sp.) is inoculated in the BM culture medium containing 100mg/L Zn (II), is surveyed
Try its aerobic denitrification capability.The growth of the bacterial strain is significantly inhibited, and nitrate nitrogen is hardly removed, it is possible thereby to
Find out, 100mg/L Zn (II) is the upper concentration of the bacterial strain, and bacterial strain does not have the ability of aerobic denitrification under this condition.But
It is from the biological treatment that can be seen that in above-described embodiment 1-4 by the strain inoculated to the sewage containing zinc, in 0~50mg/L Zn
(II) under existence condition, the removal rate of nitrate nitrogen illustrates that there is bacterial strain stronger tolerance zinc ion toxicity to become reconciled up to 100%
Oxygen denitrifying capacity (is shown in Table 1).
Nitrogen removal characteristics of 1 achromobacter of table under various concentration Zn (II) existence condition
The aerobic denitrification of 6. achromobacter of embodiment and Pseudomonas stutzeri under 50mg/L Zn (II) existence condition is special
Property
By achromobacter (Achromobacter sp.) and Pseudomonas stutzeri (P.stutzeri) be inoculated in containing
In the BM culture medium of 50mg/LZn (II), its aerobic denitrification capability is tested.The results are shown in Table 2, and 100mg/L nitrate nitrogen exists
It can be utilized immediately by colourless bar bacillus after inoculation, the maximum removal rate of nitrate nitrogen is 5.8 ± 0.5mg/L/h, and finally produces
Object is N2, illustrate that denitrification process is not suppressed.And Pseudomonas stutzeri is under the conditions of 50mg/L Zn (II) is existing, most
Big nitrate nitrogen removal rate is only 2.9 ± 0.4mg/L/h, although final denitrification product is also nitrogen, rate is only nothing
The half of color bacillus.It is well known that Pseudomonas stutzeri is typical denitrification bacterial strain, by comparison it is not difficult to find that the present invention
The achromobacter of proposition is denitrifying in high-efficiency aerobic meanwhile, it is capable to have higher tolerance (being shown in Table 2) to Zn (II).
The aerobic denitrification characteristic ratio of 2 achromobacter of table and Pseudomonas stutzeri under 50mg/L Zn (II) existence condition
Compared with
Application of 7. achromobacter of embodiment in zinc-containing water
Achromobacter is added into activated Sludge System Treatment of Zinc-containing Wastewater, it is 6mg/ that continuous aeration, which keeps system dissolved oxygen,
L, waste water quality are as follows: pH 7.5, nitrate nitrogen content 100mg/L, Zn (II) 50mg/L.The results are shown in Table 2, common
In activated Sludge System, nitrogen removal rate is lower under aerobic condition；50mg/L Zn (II) aerobic nitrogen removal performance is added and receives shadow
It rings.However, the system after achromobacter is strengthened, the nitrate nitrogen removal rate of 36h is 96.4%, nitrogen removal rate 90.5%
(being shown in Table 3).
3. achromobacter of table is applied to the biological denitrificaion result in activated sludge system
It can be seen that Zn (II) and nitrate nitrogen coexist, institute of the present invention for the problem that zinc-containing water enters sanitary sewage
The influence that the bacterial strain being related to can effectively avoid zinc ion from removing nitrate nitrogen, and can be widely used at sewage
In reason, zinc ion can be effectively cut down to the potential risk of bio-denitrifying sewage process, to ensure town sewage treatment system just
Often operation is of great significance.
It should be noted that above embodiments are intended merely to, the present invention is described in further detail, is not intended to pair
What the present invention was defined, in the range for not departing from design and spirit of the invention, those of ordinary skill in the art, Ke Yijin
The various improvement of row or variation, still fall within the protection scope of the appended claims of the present invention.
Claims (9)
1. one plant tolerance zinc ion toxicity achromobacter application, it is characterised in that: by one plant of aerobic denitrifying bacteria without
The Nitrogen removal effect of color bacillus (Achromobacter sp.) enhanced sewage processing system, can cut down depositing for excessive zinc ion
In the influence to sewage treatment biological denitrification process.
2. the application of achromobacter (Achromobacter sp.) as described in claim 1, it is characterised in that: the bacterial strain
Deposit number is CGMCC No.2964, has preferable tolerance to the zinc ion of various concentration level.
3. the application of achromobacter (Achromobacter sp.) according to claim 1, it is characterised in that: the bacterial strain
Aerobic denitrification can be carried out by carbon source of organic matter in the case where different zinc ion concentrations are horizontal using nitrate nitrogen as nitrogen source,
To which nitrate nitrogen be removed.
4. application of the achromobacter (Achromobacter sp.) as described in claim 1 in bio-denitrifying sewage, special
Sign is: achromobacter (Achromobacter sp.) is added in sewage containing zinc, and adds proper amount of carbon source and is aerated,
The removal of nitrate from waste water nitrogen can be realized.
5. application according to claim 4, it is characterised in that: controlling the temperature in the sewage containing zinc is 30 DEG C.
6. application according to claim 4, it is characterised in that: controlling the pH value in the sewage containing zinc is 7.5.
7. application according to claim 4, it is characterised in that: controlling the dissolved oxygen in the sewage containing zinc is 6mg/L.
8. application according to claim 4, it is characterised in that: controlling the initial C/N ratio in the sewage containing zinc is 4.
9. application according to claim 4, it is characterised in that: zinc ion concentration range contained by sewage is 0~50mg/L,
The removal rate of nitrate nitrogen is up to 100% under aerobic environment, and the removal rate of total nitrogen is also up to 100%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257291A (en) * | 2019-06-25 | 2019-09-20 | 北京大学 | One plant of achromobacter for being resistant to nickel ion toxicity and its application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257291A (en) * | 2019-06-25 | 2019-09-20 | 北京大学 | One plant of achromobacter for being resistant to nickel ion toxicity and its application |
| CN110257291B (en) * | 2019-06-25 | 2021-07-23 | 北京大学 | Achromobacter capable of tolerating nickel ion toxicity and application thereof |
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