CN107164275A - One plant tolerance nano titanium oxide toxicity Pseudomonas stutzeri and its application - Google Patents
One plant tolerance nano titanium oxide toxicity Pseudomonas stutzeri and its application Download PDFInfo
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- CN107164275A CN107164275A CN201710474775.6A CN201710474775A CN107164275A CN 107164275 A CN107164275 A CN 107164275A CN 201710474775 A CN201710474775 A CN 201710474775A CN 107164275 A CN107164275 A CN 107164275A
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Abstract
The present invention relates to one plant of Pseudomonas stutzeri for being resistant to nano titanium oxide toxicity and its application.One plant of Pseudomonas stutzeri (P.stutzeri) with nanometer titanium dioxide carbon tolerance has been grown in present invention training, and its preserving number is CGMCC No.14062.When there is the nano titanium oxide of high concentration level (0 128mg/L) in waste water, the bacterial strain can be using nitrate nitrogen as only nitrogen source, the clearance of nitrate nitrogen is up to 100% under aerobic environment, the clearance of total nitrogen is same up to 85%, the stronger tolerance to nano titanium oxide toxicity is shown, the influence to optimization sewage biological treatment system and decrease nano material to sewage denitrification system is significant.
Description
Technical field
The present invention relates to a kind of Pseudomonas stutzeri, more particularly to one kind is resistant to nano-silica under aerobic condition
Change titanium toxicity and can be by the Pseudomonas stutzeri of the nitrate nitrogen simultaneous removing in waste water and its in sewage disposal should
With.
Background technology
The particle diameter of nano material is typically in the range of between 1-100nm, because its special physicochemical characteristics (such as non-migratory,
Fluorescence, piezoelectricity and scatters ultraviolet ability etc.) and its marvellous performance in terms of light, electricity, magnetic, it is widely used in
The fields such as medicine, cosmetics, sterilization and anticorrosion, composite, food anti-bacterial packaging film, regenerative medicine, medicine equipment.Nanometer
TiO2It is current most popular metal oxide nanoparticles, TiO according to statistics2Annual worldwide production total amount up to 10000 tons,
About 50-80% nano-TiO2It is used for cosmetics and sunscreen product.With nano-TiO2Industrialized production and extensive use,
It is inevitably discharged into sewage collection system, and final as sanitary sewage and industrial wastewater system enter sewage treatment plant.
It is well known that municipal sewage plant generally realizes going for sewage nitrogen and phosphorus using activated sludge process both at home and abroad
Remove.In active sludge processing system, the activated sludge using microorganism as core is the important analyst of pollutant, and it grows
The effect of sewage disposal will be directly affected with the height of metaboilic level.Research shows that the presence of nano material can significantly inhibit work
The growth metabolism of microorganism especially denitrification microorganism in property sludge, so that the denitrification effect of sewage can be influenceed, to sewage disposal
The stable operation of factory brings potential hazard.
Alleviating or cut down at present influence of the nano material to biological denitrification system mainly has two ways.One kind is by logical
Cross and change the architectural characteristic of nano material to realize.However, the doping or modification of nano material can change nano material itself
Characteristic, this can produce certain influence to applications to nanostructures scope.Another way is that discovery and training are grown with nanometer material
Expect the functional microorganism of tolerance, this mode does not change the characteristic of nano material, and can make whole Sewage Biological Treatment system
System has preferable treatment effect and stronger impact resistance, is a kind of more feasible method, but in practical application mistake
Cheng Zhong, qualified microorganism is more rare, it is difficult to meet growing real needs.
Therefore, inventor considers, if it is possible to filter out resistance nano material bio-toxicity ability denitrification functions bacterium, its
Using will effectively cut down potential risk of the nano material to bio-denitrifying sewage process.
The content of the invention
It is an object of the invention to provide a kind of high-efficiency aerobic denitrogenation bacterial strain for being resistant to nano titanium oxide, pass through the bacterial strain
The Nitrogen removal effect of enhanced sewage processing system, to cut down the presence of nano titanium oxide to sewage disposal biological denitrificaion mistake
The influence of journey.
Bacterial strain provided by the present invention, is one plant of Pseudomonas stutzeri (P.stutzeri), and being one plant has single good
The aerobic denitrifying bacteria of the efficient removal ability of nitrate nitrogen is realized in oxygen environment.
The technical scheme is that：
Pseudomonas stutzeri (the Pseudomonas with nanometer titanium dioxide carbon tolerance that the present invention is provided
Stutzeri), China Committee for Culture Collection of Microorganisms's common micro-organisms center's (letter is preserved on April 24th, 2017
Claim CGMCC), address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, its preserving number
For CGMCC No.14062.
Pseudomonas stutzeri (P.stutzeri) as described above, it is characterised in that there is nano titanium oxide in waste water
Under the conditions of, nano particle can be adsorbed in phage surface, and bacterial strain P.stutzeri can keep complete eucaryotic cell structure, cell membrane
It is not destroyed.
Pseudomonas stutzeri (P.stutzeri) as described above, it is characterised in that there is receiving for low concentration level in waste water
Under conditions of rice titanium dioxide (<4mg/L), the bacterial strain can be using nitrate nitrogen as only nitrogen source, the nitrate under aerobic environment
The clearance of nitrogen is up to 100%, and the clearance of total nitrogen is up to more than 85%.
Pseudomonas stutzeri (P.stutzeri) as described above, it is characterised in that there is receiving for low concentration level in waste water
Under conditions of rice titanium dioxide (4-128mg/L), the bacterial strain can be using nitrate nitrogen as only nitrogen source, the nitre under aerobic environment
The clearance of hydrochlorate nitrogen is up to 100%, and the clearance of total nitrogen is same up to 100%.
The Pseudomonas stutzeri of the present invention and its application have the advantages that compared with prior art：
(1) according to the unique physiological characteristic of the false unit cell of the Amur of discovery, realize to the resistance to of nano titanium oxide toxicity
By solving potential impact of the presence to biological denitrification process of nano material.
(2) using the present invention, by by the secondary biochemical treatment system of the inoculation to traditional activated sludge process,
Nano oxidized zinc concentration is under 0~150mg/L existence condition, it is preferred that under 0~128mg/L existence conditions, nitrate nitrogen
Clearance up to 100%, illustrate bacterial strain have stronger tolerance nano material toxicity and aerobic denitrifying capacity.This spy
Property causes the bacterial strain to be greatly enhanced in the practicality of town sewage treatment system.
(3) present invention is convenient flexibly, has a extensive future, with good social benefit.
With reference to embodiment, the present invention will be described in detail.One of ordinary skill in the art can manage
Solution, the purpose that embodiment is merely illustrative, the scope of the present invention is not limited with specific implementation method, but by right
It is required that scope be limited.
Brief description of the drawings
Fig. 1：Fig. 1 is scanning electron microscope image of the Pseudomonas stutzeri under nano titanium oxide existence condition.
Fig. 2：Fig. 2 is nitrogen removal characteristics of the Pseudomonas stutzeri under 1mg/L nano titanium oxide existence conditions.
Fig. 3：Fig. 3 is nitrogen removal characteristics of the Pseudomonas stutzeri under 4mg/L nano titanium oxide existence conditions.
Fig. 4：Fig. 4 is nitrogen removal characteristics of the Pseudomonas stutzeri under 16mg/L nano titanium oxide existence conditions.
Fig. 5：Fig. 5 is nitrogen removal characteristics of the Pseudomonas stutzeri under 64mg/L nano titanium oxide existence conditions.
Fig. 6：Fig. 6 is nitrogen removal characteristics of the Pseudomonas stutzeri under 128mg/L nano titanium oxide existence conditions.
Embodiment
The present invention is described in further detail with embodiment below in conjunction with the accompanying drawings：
Embodiment 1
First, the mode of appearance using scanning electron microscopic observation Pseudomonas stutzeri in the presence of nano titanium oxide
High-efficiency aerobic denitrification bacterial strain is Pseudomonas stutzeri P.stutzeri used in this experiment, and it is isolated from garbage loading embeading
Field percolation liquid treating system.
Pseudomonas stutzeri P.stutzeri is inoculated in 1L KNO containing 0.5g3With 0.35g NH4In Cl LB culture mediums
(every liter of 5g containing NaCl, tryptone 10g, yeast extract 5g), prevents the intrusion of miscellaneous bacteria and keeps the growth vigor of thalline,
Carry out enrichment culture.Obtained bacterium solution centrifugation will be cultivated, washed with 0.5% NaCl three times, optical density OD is made600For 1-2
Bacteria suspension.Then bacteria suspension is inoculated in containing 16mg/L nano-TiOs by 10% inoculum concentration again2Culture medium in, after 8h
Sampling centrifuges 5min under the conditions of 8000rpm and 4 DEG C and removes supernatant, and is cleaned twice with PBS (phosphate buffer).With
2.5% glutaraldehyde is dispersed in absolute ethyl alcohol after fixing 1h, ethanol gradient elution, takes a small amount of drop on silicon chip to dry, in scanning
Cellular morphology is observed under Electronic Speculum (SEM).
As a result as shown in figure 1, working as nano-TiO2When concentration is 16mg/L, bacterial strain P.stutzeri presents complete and smooth
Surface, illustrate strain cell form to nano-TiO2With stronger tolerance.
Embodiment 2
2nd, nitrogen removal characteristics of the Pseudomonas stutzeri under 1mg/L nano titanium oxide existence conditions
Bacteria suspension after enrichment culture is inoculated in containing 1mg/L nano-TiOs according to 10% inoculum concentration2BM culture
In base, 2mL is sampled every 4h, sampling centrifuges 5min under the conditions of 8000rpm and 4 DEG C, and supernatant is used for various nitrogen-containing compounds
The measure of concentration.Denitrification capability test media (BM) formula is in every liter of water：8.45g CH3COONa,0.61g NaNO3,
1.76g K2HPO4·3H2O,0.20g MgSO4·7H2O,0.02g CaCl2, 0.005g FeSO4·7H2The micro member of O, 0.1mL
Plain solution.Medium's PH Value is adjusted to 7.5, sterilize 30min at 121 DEG C.
As shown in Fig. 2 as nano TiO2Concentration be 1mg/L when, nitrate nitrogen can all be removed in 28h.
In the removal process of nitrate nitrogen, with the accumulation of nitrite nitrogen, reached most in the accumulation of 16h nitrite nitrogens
Greatly, concrete numerical value is 65.7mg/L.With proceeding for reaction, nitrite nitrogen is gradually degraded by Pseudomonas stutzeri,
Concentration is reduced to 8.0mg/L after 48h.With the total nitrogen in the aggregate surrogates system of nitrate nitrogen and nitrite nitrogen, at this
Under part, the clearance of total nitrogen illustrates that bacterial strain implements pseudomonad to nano-TiO up to 92.4%2There is preferable tolerance.
Embodiment 3
3rd, nitrogen removal characteristics of the Pseudomonas stutzeri under 4mg/L nano titanium oxide existence conditions
Bacteria suspension after enrichment culture is inoculated in containing 4mg/L nano-TiOs according to 10% inoculum concentration2BM culture
In base, 2mL is sampled every 4h, sampling centrifuges 5min under the conditions of 8000rpm and 4 DEG C, and supernatant is used for various nitrogen-containing compounds
The measure of concentration.
As shown in figure 3, working as nano-TiO2Concentration be 4mg/L when, nitrate nitrogen can all be removed in 28h.In nitre
In the removal process of hydrochlorate nitrogen, with the accumulation of nitrite nitrogen, maximum is reached in the accumulation of 20h nitrite nitrogens, is had
Body numerical value is 64.8mg/L.With proceeding for reaction, nitrite nitrogen is gradually degraded by Pseudomonas stutzeri, after 48h
Concentration is reduced to 16.2mg/L.With the total nitrogen in the aggregate surrogates system of nitrate nitrogen and nitrite nitrogen, in this condition
Under, the clearance of total nitrogen is up to 84.6%.Nitrogen removal rate under this concentration level is slightly less than nano-TiO2Concentration level
Situation during for 1mg/L, but bacterial strain Pseudomonas stutzeri is still able to efficient removal of the realization to nitrogen.
Embodiment 4
4th, nitrogen removal characteristics of the Pseudomonas stutzeri under 16mg/L nano titanium oxide existence conditions
Bacteria suspension after enrichment culture is inoculated in containing 16mg/L nano-TiOs according to 10% inoculum concentration2BM culture
In base, 2mL is sampled every 4h, sampling centrifuges 5min under the conditions of 8000rpm and 4 DEG C, and supernatant is used for various nitrogen-containing compounds
The measure of concentration.
As shown in figure 4, working as nano-TiO2Concentration when being 16mg/L, nitrate nitrogen can be in 20h clearances up to 100%.
In the removal process of nitrate nitrogen, with the accumulation of nitrite nitrogen, reached most in the accumulation of 12h nitrite nitrogens
Greatly, concrete numerical value is 65.3mg/L.With proceeding for reaction, nitrite nitrogen is gradually degraded by Pseudomonas stutzeri,
Nitrite is completely removed after 48h.With the total nitrogen in the aggregate surrogates system of nitrate nitrogen and nitrite nitrogen, at this
Under the conditions of, the clearance of total nitrogen is up to 100%.Illustrate the nano-TiO in high concentration level2Under the conditions of, bacterial strain Amur vacation unit cell
Bacterium is to nano-TiO2With stronger tolerance.
Embodiment 5
5th, nitrogen removal characteristics of the Pseudomonas stutzeri under 64mg/L nano titanium oxide existence conditions
Bacteria suspension after enrichment culture is inoculated in containing 64mg/L nano-TiOs according to 10% inoculum concentration2BM culture
In base, 2mL is sampled every 4h, sampling centrifuges 5min under the conditions of 8000rpm and 4 DEG C, and supernatant is used for various nitrogen-containing compounds
The measure of concentration.
As shown in figure 5, working as nano-TiO2Concentration when being 64mg/L, nitrate nitrogen can be in 16h clearances up to 100%.
In the removal process of nitrate nitrogen, with the accumulation of nitrite nitrogen, reached most in the accumulation of 12h nitrite nitrogens
Greatly, concrete numerical value is 65.8mg/L.With proceeding for reaction, nitrite nitrogen is gradually degraded by Pseudomonas stutzeri,
Nitrite is completely removed after 24h.With the total nitrogen in the aggregate surrogates system of nitrate nitrogen and nitrite nitrogen, at this
Under the conditions of, the clearance of total nitrogen is up to 100%.Illustrate bacterial strain Pseudomonas stutzeri to highly concentrated nano TiO2With stronger
Tolerance.
Embodiment 6
6th, nitrogen removal characteristics of the Pseudomonas stutzeri under 128mg/L nano titanium oxide existence conditions
Bacteria suspension after enrichment culture is inoculated in containing 128mg/L nano-TiOs according to 10% inoculum concentration2BM training
Support in base, 2mL is sampled every 4h, sampling centrifuges 5min under the conditions of 8000rpm and 4 DEG C, and supernatant is used for various nitrogenous chemical combination
The measure of thing concentration.
As shown in figure 5, working as nano-TiO2Concentration be 128mg/L when, nitrate nitrogen can be completely removed in 10h.
In the removal process of nitrate nitrogen, with the accumulation of nitrite nitrogen, reached most in the accumulation of 12h nitrite nitrogens
Greatly, concrete numerical value is 65.8mg/L.With proceeding for reaction, nitrite nitrogen is gradually degraded by Pseudomonas stutzeri,
Nitrite is completely removed after 20h.With the total nitrogen in the aggregate surrogates system of nitrate nitrogen and nitrite nitrogen, at this
Under the conditions of, the clearance of total nitrogen is up to 100%.Illustrate that concentration is higher, bacterial strain Pseudomonas stutzeri is to highly concentrated nano TiO2's
Tolerance is stronger (being shown in Table 1).
Nitrogen removal characteristics of the Pseudomonas stutzeri of table 1 under various concentrations nano titanium oxide existence condition
Embodiment 7
7th, application of the Pseudomonas stutzeri in waste water containing nano titanium oxide
Pseudomonas stutzeri is added and handles waste water containing nano titanium oxide, continuous aeration keeping body into activated Sludge System
It is that dissolved oxygen is 6mg/L, waste water quality is as follows：PH is 7.5, nitrate nitrogen content 100mg/L, nano titanium oxide 1mg/L.
In ordinary activated sludge system, the effect of denitrogenation receives large effect under conditions of nano titanium oxide presence.So
And, the system after strengthening through Pseudomonas stutzeri, 24h nitrate nitrogen clearance is up to 79.3%, and nitrogen removal rate is reachable
61.7%；36h nitrate nitrogen clearance is 96.4%, and nitrogen removal rate is 90.1% (being shown in Table 2).
The Pseudomonas stutzeri PCN-1 of table 2 is applied to the biological denitrificaion result in activated sludge system
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 (5)
1. the application of the Pseudomonas stutzeri of one plant of tolerance nano titanium oxide toxicity, it is characterised in that：It is aerobic anti-by one plant
The Nitrogen removal effect of nitrobacteria Pseudomonas stutzeri (P.stutzeri) enhanced sewage processing system, can cut down nano-silica
Change influence of the presence of titanium to sewage disposal biological denitrification process.
2. application as claimed in claim 1, wherein the preserving number of described Pseudomonas stutzeri (P.stutzeri) is CGMCC
No.14062, to the nano-TiO of various concentrations level2With preferable tolerance.
3. the application of Pseudomonas stutzeri (P.stutzeri) according to claim 1, it is characterised in that：When being deposited in waste water
In nano titanium oxide, nano particle absorption can be kept complete eucaryotic cell structure by the bacterial strain while phage surface,
Cell membrane is not destroyed.
4. the application of Pseudomonas stutzeri (P.stutzeri) according to claim 1, it is characterised in that：When being deposited in waste water
Low concentration level (<During nano titanium oxide 4mg/L), the bacterial strain can be using nitrate nitrogen as only nitrogen source, in aerobic ring
The clearance of nitrate nitrogen is up to 100% under border, and the clearance of total nitrogen is up to more than 85%.
5. the application of Pseudomonas stutzeri (P.stutzeri) according to claim 1, it is characterised in that exist in waste water
During the nano titanium oxide of high concentration level (4-128mg/L), the bacterial strain can be using nitrate nitrogen as only nitrogen source, in aerobic ring
The clearance of nitrate nitrogen is up to 100% under border, and the clearance of total nitrogen is same up to 100%.
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| CN110104768A (en) * | 2019-04-18 | 2019-08-09 | 杭州师范大学 | A kind of cultural method for the denitrification sludge being resistant to nano material |
| 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 |
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| CN110104768B (en) * | 2019-04-18 | 2021-12-21 | 杭州师范大学 | Culture method of nano-material-tolerant denitrifying sludge |
| 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 |
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| CN107164275B (en) | 2020-07-31 |
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