CN110452846A - A kind of Paracoccus denitrificans and its method with mineralising bed combination progress biological denitrificaion - Google Patents
A kind of Paracoccus denitrificans and its method with mineralising bed combination progress biological denitrificaion Download PDFInfo
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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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- 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
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- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F2003/001—Biological treatment of water, waste water, or sewage using granular carriers or supports for the microorganisms
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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
- C02F2003/001—Biological treatment of water, waste water, or sewage using granular carriers or supports for the microorganisms
- C02F2003/003—Biological treatment of water, waste water, or sewage using granular carriers or supports for the microorganisms using activated carbon or the like
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F2101/00—Nature of the contaminant
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- C02F2101/16—Nitrogen compounds, e.g. ammonia
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/02—Temperature
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/06—Controlling or monitoring parameters in water treatment pH
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2301/00—General aspects of water treatment
- C02F2301/10—Temperature conditions for biological treatment
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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
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Abstract
The present invention provides a kind of Paracoccus denitrificans and its method with mineralising bed combination progress biological denitrificaion, the present invention provides one plant of autotrophic denitrification Paracoccus denitrificans TNS-1, and select mineralized waste as packed bed main component, realize the resource utilization of waste, total nitrogen removing is carried out using bioaugmentation, it is environmental-friendly, it is without secondary pollution, processing cost is lower, is a kind of denitrogenation of waste water method of low energy consumption, environmentally friendly low-carbon.
Description
Technical field
The present invention relates to a kind of Paracoccus denitrificans, carry out biology the invention further relates to Paracoccus denitrificans and the combination of mineralising bed and take off
The method of nitrogen belongs to sewage treatment field.
Background technique
Mineralized waste refers to that in refuse landfill after the several years degrade, degradable material completely or nearly drops completely
Solution, basically reaches stable state, and surface settlement amount is very small, and spontaneous percolate and gas flow are few or do not generate,
Biodegradable material mass fraction drops to 3% or less.On appearance composition, except plastics, glass, stone, animal skeleton etc.
Outside bulky grain inorganic matter and hardly degraded organic substance, remaining is homogeneous black discrete particles shape class soil material.Mineralising rubbish
Rubbish has good cellular structure, huge surface area, and is attached with microbial population abundant, is a kind of good biology
Filler.
Since the nineties in last century, numerous scholars have started the further investigation to mineralized waste, as a result show mineralising
Rubbish has good treatment effect to ammonia nitrogen, but the problem of not can solve effluent nitrate-nitrogen too high levels, causes to be discharged
Total nitrogen is high.One of its reason is that conventional heterotrophic denitrification process, need to be additionally provided organic matter as carbon source and
Electron donor will hinder the denitrification process of nitrate nitrogen when carbon source deficiency carbon-nitrogen ratio is too low in system.
Paracoccus denitrificans is a kind of Gram-negative bacteria of the inorganic autotrophy of facultativeization energy, it can utilize various carbohydrates, ammonia
The organic matters such as base acid carry out heterotrophic growth, also can be in H2、O2、CO2, carry out autophyting growth in the presence of the inorganic salts of part.
In the presence of in system without molecular oxygen, Paracoccus denitrificans can using nitrate or nitrite as final electron acceptor,
Carry out denitrification denitrogenation process.
Summary of the invention
In order to solve the problems, such as that it is conventional denitrifying that low-carbon high-nitrogen waste water is difficult to, and mineralized waste is rationally utilized, the present invention mentions
One plant of autotrophic denitrification Paracoccus denitrificans TNS-1 has been supplied, it has been provided and expands cultural method, and it is joined with mineralized waste packed bed
With the removing of realization high-nitrogen waste water total nitrogen.
Present invention employs following technical solutions:
The present invention provides a kind of Paracoccus denitrificans, it is characterised in that: Paracoccus denitrificans
(Paracoccus.denitrificans) TNS-1, deposit number are CGMCC NO:17156.
The present invention also provides the methods that Paracoccus denitrificans and the combination of mineralising bed carry out biological denitrificaion characterized by comprising
Step 1: Spawn incubation
First cell culture medium: 7.0,121 DEG C of high pressure steam sterilization 20min of peptone 5.0g/L, potassium nitrate 1.0g/L, pH,
Secondary medium: Na2S2O3·5H2O 5.0, KNO31.0g/L, K2HPO42.0g/L, NaHCO31.0g/L
MgCl2·6H2O 0.5g/L, FeSO4·7H2O 0.01g/L, is adjusted to predetermined pH,
Paracoccus denitrificans (Paracoccus.denitrificans) TNS-1 strain after taking activation, deposit number are
CGMCC NO:17156, by the sterile access first cell culture medium of 1% inoculum concentration, 30 DEG C, 120r/min shaken cultivation 24-48h takes one
Grade strain spreads cultivation in secondary medium of transferring by 1-2% inoculum concentration, and stationary culture 5-7d obtains second level and spreads cultivation bacterium solution；
Step 2: packed bed preparation
Mineralized waste pretreatment: mineralized waste sieving removes the large particulate matters such as contained glass, metal, plastics, stone, obtains
It is less than the soil shape material of 30mm to partial size,
Denitrogenation packed bed component, in mass ratio: mineralized waste 60%, drusen 20%, granular activated carbon 20% will be with
Upper component mixes in proportion, and is packed into filling tank,
Step 3: strain absorption
Bacterium solution pours into filling tank after spreading cultivation, until just flood denitrogenation packed bed top layer, keep floodage 3-5 days to
Strain is adsorbed in filler surface naturally, and the filling tank after the completion of strain absorption is referred to as denitrogenation filling tank,
Step 4: oxygen removal
High-nitrogen waste water first passes through the filtrate tank for being filled with sponge iron before entering denitrogenation filling tank, removes molecular oxygen,
Make be discharged ORP be maintained at -100mv hereinafter,
Step 5: denitrogenation operation
In denitrogenation filling tank, equivalent waste water is passed in and out respectively daily.
The method that Paracoccus denitrificans of the invention and the combination of mineralising bed carry out biological denitrificaion, also has a feature in that it
In, the temperature of the denitrogenation operation of step 5 is carried out at 10-50 DEG C.
The method that Paracoccus denitrificans of the invention and the combination of mineralising bed carry out biological denitrificaion, also has a feature in that it
In, the temperature of the denitrogenation operation of step 5 carries out under the conditions of 30 DEG C.
The method that Paracoccus denitrificans of the invention and the combination of mineralising bed carry out biological denitrificaion, also has a feature in that step
In rapid one, the predetermined pH is 3.0-11.0.
The method that Paracoccus denitrificans of the invention and the combination of mineralising bed carry out biological denitrificaion, also has a feature in that step
In rapid one, the predetermined pH is 6.0-8.0.
Advantageous effect of the invention
The present invention provides one plant of autotrophic denitrification Paracoccus denitrificans TNS-1, and select mineralized waste as packed bed master
Component is wanted, realizes the resource utilization of waste, total nitrogen removing is carried out using bioaugmentation, it is environmental-friendly, without secondary dirt
Dye, processing cost is lower, is a kind of denitrogenation of waste water method of low energy consumption, environmentally friendly low-carbon.
Detailed description of the invention
Fig. 1 (a) is influence of the different temperatures to growth；
Fig. 1 (b) is influence of the different pH to growth；
Fig. 2 (a) is influence of the different temperatures to denitrogenation；
Fig. 2 (b) is influence of the different pH to denitrogenation；
Fig. 3 is the result of lab scale total nitrogen removal；
Fig. 4 is the result of pilot scale total nitrogen removing.
It is micro- that Paracoccus denitrificans (Paracoccus.denitrificans) TNS-1 is preserved in China on January 10th, 2019
Biological inoculum preservation administration committee common micro-organisms center (address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese section
Institute of microbiology, institute, postcode: 100101), deposit number is CGMCC NO:17156.
Specific embodiment
Illustrate a specific embodiment of the invention below in conjunction with attached drawing.
Embodiment 1
The cultural method of Paracoccus denitrificans TNS-1 provided by the invention is determined by following steps:
1, actication of culture
Seed culture medium (g/L): peptone 5.0, potassium nitrate 1.0, pH 7.0.121 DEG C of high pressure steam sterilization 20min.
Denitrogenation culture medium (g/L): Na2S2O3·5H2O 5.0, KNO31.0, K2HPO42.0, NaHCO31.0, MgCl2·
6H2O 0.5, FeSO4·7H2O 0.01, PH 8.0-8.5.121 DEG C of high pressure steam sterilization 20min.
It goes bail for Paracoccus denitrificans (Paracoccus.denitrificans) the TNS-1 slant strains deposited, under aseptic condition,
In seed culture medium after being inoculated with a ring to sterilizing, 30 DEG C, 120r/min shaken cultivation 24-48h, as strain.
2, growth conditions is tested
1) influence of the different temperatures to growth
Strain access seed culture medium after activating, inoculum concentration 1%, 30 DEG C, 120r/min shaken cultivation.10 are selected respectively
DEG C, 20 DEG C, 30 DEG C, 40 DEG C, 50 DEG C of totally 5 cultivation temperatures, measure bacterium solution absorbance change, investigate different temperatures to bacterial strain TNS-
The influence of 1 growth.As a result as shown in Fig. 1 (a), what Paracoccus denitrificans (Paracoccus.denitrificans) TNS-1 was suitable for
Growth temperature is 30-40 DEG C.
2) influence of the difference pH to growth
Strain access seed culture medium after activating, inoculum concentration 1%, 30 DEG C, 120r/min shaken cultivation.It is set separately
PH3.0,4.0,5.0,6.0,7.0,8.0,9.0,10.0,11.0, totally 9 pH conditions measure bacterium solution absorbance change, investigate training
The influence that nutrient solution difference pH value grows bacterial strain TNS-1.As a result as shown in Fig. 1 (b), Paracoccus denitrificans
(Paracoccus.denitrificans) TNS-1 suitable growth in neutral environment, optimal pH range is 6.0-8.0.
3, denitrogenation condition test
1) different temperatures influences denitrogenation
By 1% inoculum concentration, seed is accessed into denitrogenation culture medium, selectes 10 DEG C, 20 DEG C, 30 DEG C, 40 DEG C, 50 DEG C respectively, totally 5
A cultivation temperature, stationary culture.Total nitrogen variation is measured, influence of the different temperatures to bacterial strain TNS-1 nitrogen removal performance is investigated.As a result such as
Shown in Fig. 2 (a), the optimum temperature of Paracoccus denitrificans (Paracoccus.denitrificans) TNS-1 denitrogenation is 30 DEG C.
2) different initial pH influence denitrogenation
By 1% inoculum concentration, seed is accessed into denitrogenation culture medium, be set separately the initial pH of culture medium be 3.0,4.0,5.0,
6.0,7.0,8.0,9.0,10.0,11.0, totally 9 pH conditions, 30 DEG C of stationary cultures.Total nitrogen variation is measured, is investigated different initial
Influence of the pH to bacterial strain TNS-1 nitrogen removal performance.As a result as shown in Fig. 2 (b), Paracoccus denitrificans
(Paracoccus.denitrificans) TNS-1 denitrification effect when initial pH is 8.0 is best.
Embodiment 2
Denitrogenation lab scale
1, culture medium
First cell culture medium (g/L): peptone 5.0, potassium nitrate 1.0, pH 7.0.121 DEG C of high pressure steam sterilization 20min.
Secondary medium (g/L): Na2S2O3·5H2O 5.0, KNO31.0, K2HPO42.0, NaHCO31.0, MgCl2·
6H2O 0.5, FeSO4·7H2O 0.01, pH 8.0.
2, Spawn incubation
Paracoccus denitrificans (Paracoccus.denitrificans) TNS-1 strain after activating is taken, by 1% inoculum concentration, nothing
Bacterium access first cell culture medium, 30 DEG C, 120r/min shaken cultivation 24-48h.Take first class inoculum, by 1-2% inoculum concentration, transfer into
It spreads cultivation in secondary medium, stationary culture 5-7d.
3, strain adsorbs
Bacterium solution pours into filling tank after spreading cultivation, until just flooding filler top layer, floodage 3-5d is kept to wait for strain certainly
So it is adsorbed in filler surface.
4, prepared by packed bed
Mineralized waste pretreatment: mineralized waste come from Shanghai Lao Gang household refuse landfill sites, heap age 7 years or more.It digs out
Mineralized waste sieving removes the large particulate matters such as contained glass, metal, plastics, stone, obtains the soil shape that partial size is less than 30mm
Material.
Denitrogenation packed bed component: mineralized waste 60%, drusen 20%, granular activated carbon 20%.The above component is pressed
Ratio is uniformly mixed, and is packed into tank.
5, oxygen removal
High-nitrogen waste water sample first passes through the filtrate tank for being filled with sponge iron particle before carrying out denitrogenation, removes molecule
Oxygen makes water outlet ORP be maintained at -100mv or less.
6, denitrogenation lab scale
High-nitrogen waste water water quality: CODCr344mg/L；Ammonia nitrogen 29mg/L；TN 1466mg/L.
The thin mouth glass jar of 2 5L is taken, is equipped with discharging valve close to bottom of bottle.No. 1 canned to fill out denitrogenation packed bed component, and is added de-
Nitrogen pair coccus TNS-1 second level spreads cultivation bacterium solution, makes liquid level to flooding filler surface after stablizing.No. 2 tanks only load mineralized waste material,
Bacterium solution is not added, high-nitrogen waste water after equivalent deoxidation is added, makes liquid level to filler surface is flooded after stablizing, No. 2 tanks are as control.
No. 1, No. 2 tanks daily respectively pass in and out equivalent waste water.Hydraulic detention time is about 5 days.
As a result as shown in figure 3, after stable operation, No. 1 denitrogenation tank water outlet nitrogen removal rate is about 80%.No. 2 control canisters are total
Nitrogen does not obviously remove, no denitrification effect.
Embodiment 3
Denitrogenation pilot scale
1, culture medium
First cell culture medium (g/L): peptone 5.0, potassium nitrate 1.0, pH 7.0.121 DEG C of high pressure steam sterilization 20min.
Secondary medium (g/L): Na2S2O3·5H2O 5.0, KNO31.0, K2HPO42.0, NaHCO31.0, MgCl2·
6H2O 0.5, FeSO4·7H2O 0.01, pH 8.0.
2, Spawn incubation
Paracoccus denitrificans (Paracoccus.denitrificans) TNS-1 strain after activating is taken, by 1% inoculum concentration, nothing
Bacterium access first cell culture medium, 30 DEG C, 120r/min shaken cultivation 24-48h.Take first class inoculum, by 1-2% inoculum concentration, transfer into
It spreads cultivation in secondary medium, stationary culture 5-7d.
3, strain adsorbs
Bacterium solution pours into filling tank after spreading cultivation, until just flooding filler top layer, floodage 3-5d is kept to wait for strain certainly
So it is adsorbed in filler surface.
4, prepared by packed bed
Mineralized waste pretreatment: mineralized waste come from Shanghai Lao Gang household refuse landfill sites, heap age 7 years or more.It digs out
Mineralized waste sieving removes the large particulate matters such as contained glass, metal, plastics, stone, obtains the soil shape that partial size is less than 30mm
Material.In other embodiments, heap age is not particularly limited, as long as forming mineralized waste.
Denitrogenation packed bed component: mineralized waste 60%, drusen 20%, granular activated carbon 20%.The above component is pressed
Ratio is uniformly mixed, and is packed into tank.
5, oxygen removal
High-nitrogen waste water sample first passes through the filtrate tank for being filled with sponge iron particle before carrying out denitrogenation, removes molecule
Oxygen makes water outlet ORP be maintained at -100mv or less.
6, denitrogenation pilot scale
High-nitrogen waste water water quality: CODCr344mg/L；Ammonia nitrogen 29mg/L；TN 1466mg/L.
1 50L polyethylene tank is taken, tank mouth installs water distributor, water (flow) direction upper entering and lower leaving.Denitrogenation packed bed group is loaded in tank
Point, Paracoccus denitrificans TNS-1 second level is added and spreads cultivation bacterium solution, makes liquid level to flooding filler surface after stablizing.
Daily disengaging equivalent high-nitrogen waste water.Hydraulic detention time is about 5 days.
As a result as shown in figure 4, runing time 1 month or so, it is about 75% that denitrogenation pilot scale, which is discharged total nitrogen average removal rate,.It is de-
Nitrogen effect is obvious.
Above-mentioned test duration is run two months, as a result show it is stable after, nitrogen removal rate stablizes 71~
79%, total nitrogen average removal rate is 75%.
Claims (6)
1. a kind of Paracoccus denitrificans, it is characterised in that:
Paracoccus denitrificans (Paracoccus.denitrificans) TNS-1, deposit number are CGMCC NO:17156.
2. the method that Paracoccus denitrificans and the combination of mineralising bed carry out biological denitrificaion characterized by comprising
Step 1: Spawn incubation
First cell culture medium: 7.0,121 DEG C of high pressure steam sterilization 20min of peptone 5.0g/L, potassium nitrate 1.0g/L, pH,
Secondary medium: Na2S2O3·5H2O 5.0, KNO31.0g/L, K2HPO42.0g/L, NaHCO31.0g/L, MgCl2·
6H2O 0.5g/L, FeSO4·7H2O 0.01g/L, is adjusted to predetermined pH,
Paracoccus denitrificans (Paracoccus.denitrificans) TNS-1 strain after taking activation, deposit number CGMCC
NO:17156, by the sterile access first cell culture medium of 1% inoculum concentration, 30 DEG C, 120r/min shaken cultivation 24-48h takes level-one bacterium
Kind, it by 1-2% inoculum concentration, spreads cultivation in secondary medium of transferring, stationary culture 5-7d obtains second level and spreads cultivation bacterium solution；
Step 2: packed bed preparation
Mineralized waste pretreatment: mineralized waste sieving removes the large particulate matters such as contained glass, metal, plastics, stone, obtains grain
Diameter is less than the soil shape material of 30mm,
Denitrogenation packed bed component, in mass ratio: mineralized waste 60%, drusen 20%, granular activated carbon 20%, by above group
Divide and mix in proportion, is packed into filling tank,
Step 3: strain absorption
Bacterium solution pours into filling tank after spreading cultivation, until just flooding denitrogenation packed bed top layer, holding floodage 3-5 days to strain
Naturally it is adsorbed in filler surface, the filling tank after the completion of strain absorption is referred to as denitrogenation filling tank,
Step 4: oxygen removal
High-nitrogen waste water first passes through the filtrate tank for being filled with sponge iron before entering denitrogenation filling tank, removes molecular oxygen, uses
Water ORP be maintained at -100mv hereinafter,
Step 5: denitrogenation operation
In denitrogenation filling tank, equivalent waste water is passed in and out respectively daily.
3. the method that Paracoccus denitrificans as claimed in claim 2 and the combination of mineralising bed carry out biological denitrificaion, it is characterised in that:
Wherein, the temperature to spread cultivation in step 1 is carried out at 30-40 DEG C.
4. the method that Paracoccus denitrificans as claimed in claim 2 and the combination of mineralising bed carry out biological denitrificaion, it is characterised in that:
Wherein, the temperature of the denitrogenation operation of step 5 carries out under the conditions of 30 DEG C.
5. the method that Paracoccus denitrificans as claimed in claim 2 and the combination of mineralising bed carry out biological denitrificaion, it is characterised in that:
In step 1, the predetermined pH is 3.0-11.0.
6. the method that Paracoccus denitrificans as claimed in claim 2 and the combination of mineralising bed carry out biological denitrificaion, it is characterised in that:
In step 1, the predetermined pH is 6.0-8.0.
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| CN111233141B (en) * | 2020-03-02 | 2021-08-27 | 同济大学 | Method for promoting microbial denitrification |
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