CN110144308B - High-salt-tolerance denitrifying bacterium capable of efficiently degrading nitrate, and preparation and application thereof - Google Patents

High-salt-tolerance denitrifying bacterium capable of efficiently degrading nitrate, and preparation and application thereof Download PDF

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CN110144308B
CN110144308B CN201910396425.1A CN201910396425A CN110144308B CN 110144308 B CN110144308 B CN 110144308B CN 201910396425 A CN201910396425 A CN 201910396425A CN 110144308 B CN110144308 B CN 110144308B
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任仁
胡玖坤
晋超
苗时雨
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Dajiang Environment Co ltd
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Abstract

The invention discloses a denitrifying bacterium which is tolerant to high salt and can efficiently degrade nitrate, and a preparation method and application thereof, belonging to the field of environmental microorganisms, wherein the bacterium is named as Pannonibacter phragmitetus, an anaerobic denitrifying bacterium strain, the preservation number is CGMCC No.15312, and the preservation date is 2018, 1 month and 25 days. The Pannonibacter phragmitetus has high salt-tolerant denitrification characteristic, the denitrification rate reaches 99%, and high-concentration nitrate Nitrogen (NO) in a water body can be obtained under the condition that the salt content is less than 8%3 ‑<75000mg/L) into harmless nitrogen and no nitrite accumulation, and can shorten the startup period by applying the catalyst in a denitrification reactor to realize high-efficiency and stable denitrification and no secondary pollution.

Description

High-salt-tolerance denitrifying bacterium capable of efficiently degrading nitrate, and preparation and application thereof
Technical Field
The invention belongs to the field of environmental microorganisms, and particularly relates to a high-salt-tolerance denitrifying bacterium capable of efficiently degrading nitrate, and a preparation method and application thereof.
Background
With the continuous improvement of social development and industrialization level, the environmental problem is increasingly prominent, the discharge amount of nitrogen-containing pollutants in China is rapidly increased, the nitrogen-containing pollutants are one of the main sources of water eutrophication, and the fresh water ecosystem is seriously damaged by high-nitrogen high-saline-alkali wastewater produced by industry. Wherein the aquaculture water body is seriously polluted by nitrite, the concentration of the nitrite in some aquaculture water is as high as 50mg/L or even higher, and the life metabolic activity of people and animals is seriously influenced.
According to rough statistics, about 3000 thousands of people in China drink high-nitrate saline water, and nitrate pollution becomes one of the main environmental factors for cancer occurrence in China. After being taken into human and animal bodies, part of nitrate is reduced into nitrite. Nitrite can oxidize hemoglobin in blood into methemoglobin which does not have the capacity of combining oxygen, and when the methemoglobin content in blood is increased, the oxygen conveying capacity of the blood is reduced, and serious patients cause purpura of human tissues, which is clinically called as methemoglobinemia. At present, three-stage treatment is generally adopted in national urban sewage treatment, the nitrogen and phosphorus removal effect is good, the effluent of most sewage plants can reach the first-level A discharge standard, and the continuous deterioration of water quality pollution is effectively restrained.
However, the high-concentration nitrate waste liquid generated in industrial activities has the characteristics of complex water quality components, organic pollutants, high salinity, strong biological toxicity and the like, and increases the difficulty of denitrification of the waste liquid. Waste liquid discharged from some industrial activities such as chemical fertilizer manufacturing, gunpowder manufacturing, metal surface pickling, electroplating, circuit printing plate etching and the like contains high-concentration nitrate and nitrite. For example, the content of nitrate in the waste liquid discharged by a potassium nitrate production plant reaches 2640mg/L and the content of nitrite in the waste liquid reaches 640 mg/L. For example, the concentration of nitrate contained in waste liquid generated when the rack is deplated in the electroplating industry is as high as 100000 mg/L.
Nitrogen mainly represented by NH4 +-N、NO3 --N、NO2 -The form-N, etc. exists in the ecological environment. The chemical method comprises a chemical oxidation method and a chemical reduction method, wherein the chemical oxidation method is to add an oxidant (such as hydrogen peroxide, sodium hypochlorite, ozone and the like) to oxidize nitrite into nitrate; the chemical reduction method is to add reducing agent (such as metallic iron, ferrous sulfate, hydrogen, etc.) to reduce nitrite into NH4 +And N2It may cause secondary pollution. The biological method for removing the nitrite has the advantages of high efficiency, low consumption, no secondary pollution and the like.
For example, the article, "research progress of aerobic denitrifying bacteria and action mechanism for removing nitrite" (Hetenxia, etc., proceedings of Process engineering) summarizes the types and characteristics of aerobic denitrifying bacteria and the action mechanism and way for removing nitrite, systematically analyzes the biological nitrogen removal mode, including assimilation, enzymatic degradation, acid degradation and ammonium production catabolism reduction, the denitrification reduction of nitrite is mainly an enzyme catalytic reaction, specifically, nitrite reductase, nitric oxide reductase and nitric oxide reductase, and shows relay relation to the catalytic reaction of nitrite, in the future, the research can be enhanced from the aspects of separating and screening high-efficiency nitrite type denitrifying bacteria, optimizing enzymatic reaction technology, deeply analyzing biological denitrification influencing factors and the like, and the denitrification efficiency of the denitrifying bacteria in practical application is improved.
For the denitrification of waste liquid, the conversion of nitrate nitrogen into nitrogen by the denitrification of biological method is the most economic and thorough treatment technology. However, the high salinity waste liquid discharged from industrial production can cause the osmotic pressure of the biological membrane of the microorganism in the conventional biological treatment system to be higher and the cell membrane of the microorganism to be broken; microbial enzyme activity is inhibited; the sludge settling effect becomes poor, etc. Therefore, the conventional biological denitrification process cannot effectively perform denitrification under the condition of high salt, and the processes such as ion exchange, evaporative crystallization, reverse osmosis, catalytic reduction and the like are commonly adopted in the industry for treating the high-concentration nitrate waste liquid at present, but the methods have the problems of pollution transfer, secondary pollution and the like, and the final treatment of pollutants cannot be realized. Therefore, screening of high-efficiency strains with salt tolerance is one of effective ways for solving the denitrification and denitrification of high-concentration nitrate under the high-salt condition.
Disclosure of Invention
The invention provides a denitrifying bacterium which can tolerate high salt and efficiently degrade nitrate and a preparation method and application thereof, aiming at solving the problems that the treatment of high-concentration nitrate waste liquid is not thorough, secondary pollution exists and the salt tolerance effect of the conventional denitrification process is poor in the prior art.
A denitrifying bacterium which can tolerate high-salt and efficiently degrade nitrate is Pannonibacter phragmitetus, and the preservation number is CGMCC No. 15312.
The Pannonibacter phragmitetus is preserved in the general microbiological culture Collection center of the microbiological culture Collection management Committee in 2018, 1 month and 25 months, the preservation number is CGMCC No.15312, and the preservation address is as follows: western road No.1, north chen west road, north kyo, chaoyang, institute of microbiology, china academy of sciences, zip code 100101.
The Pannonibacter phragmiteus is obtained by domesticating sludge taken from an industrial waste liquid anaerobic pool for 400 days, separating and screening, and gradually increasing the salinity and nitrate concentration of an anaerobic sludge substrate in the 400-day domestication process. The anaerobic denitrifying bacteria are suitable for separation in an enrichment separation culture medium at the temperature of 35 ℃ and the pH value of 6.6-8.0.
The 16S rDNA sequence of the Pannonibacter phragmitetus is shown in a sequence table SEQ ID No. 1.
The Pannonibacter phragmitetus strain is characterized in that: gram negative, the color of the bacterial colony is light yellow, the bacterial colony is a single convex bacterial colony, the surface of the bacterial colony is wrinkled, and the edge of the bacterial colony is irregular; the shape of the thallus is rod-shaped, straight or bent into arc shape when observed under a projection electron microscope.
The concentration of the sodium chloride salt in the waste liquid of the Pannonibacter phragmitetus is 1-10%, and the nitrate ion (NO) is endured3 -) The concentration was 2000-75000 mg/L.
The invention also aims to provide the microbial inoculum of the denitrifying bacteria which is tolerant to high salt and can efficiently degrade nitrate, wherein the microbial inoculum is specifically dry powder of the microbial inoculum or has the bacterial activity of 103-104cfu/L bacterial suspension.
The invention also aims to provide a preparation method of the denitrifying bacteria agent which is tolerant to high salt and can efficiently degrade nitrate, which comprises the following steps:
(1) fermentation culture: activating the Pannonibacter phragmititus, inoculating the activated Pannonibacter phragmitius into a fermentation culture medium according to the inoculation amount of 3-10% (volume ratio), and controlling the fermentation culture conditions as follows: 20-40 ℃, dissolved oxygen <0.2mg/L, pot pressure: shaking at 30-55rpm under 0.02-0.10Mpa for 70-96 hr; the fermentation medium comprises the following components: 6g/L-12g/L peptone, 3g/L-6g/L yeast extract, 30-60g/L sodium chloride, and the balance of distilled water, wherein the pH value is 7.5, the mixture is sterilized at 121 ℃ for 20min, and the mixture is cooled to room temperature after being sterilized;
(2) and (2) centrifuging and freeze-drying the fermentation liquor obtained in the step (1) to obtain thallus dry powder or centrifuging and diluting the thallus to obtain a thallus suspension.
Preferably, in the step (1), the activation step of the Pannonibacter phragmitetus is: the Pannonibacter phragmitius is inoculated into a seed culture medium, the culture temperature is controlled to be 30-40 ℃, and the shaking culture time is controlled to be 45-72h at 30-55 rpm.
The seed liquid culture medium comprises the following components: KNO3 2g/L-5g/L、MgSO4·7H2O 0.2g/L-0.5g/L、K2HPO40.5-0.8 g/L, 20-30 g/L of sodium potassium tartrate, 30-60g/L of sodium chloride and the balance of water, the pH value is 7.2, and the sterilization is carried out for 20min at 121 ℃.
Preferably, in the step (2), the centrifugation conditions are as follows: centrifuging at 6000-7500rpm at 4-10 deg.C for 10-15 min.
Preferably, in the step (2), the bacterial suspension is obtained by diluting the bacterial precipitate obtained by fermentation with the fermentation medium or with saline water with sodium chloride ion strength such as the fermentation medium.
And the diluting with the equal sodium chloride ion strength specifically comprises the step of diluting the thallus precipitate by using saline with the sodium chloride concentration of 30-60g/L to obtain a bacterial suspension.
More preferably, in the step (2), the bacterial suspension is specifically obtained by diluting the bacterial precipitate obtained by fermentation with the fermentation medium.
The invention also aims to provide the application of the denitrifying bacteria which are tolerant to high salt and can efficiently degrade nitrate in the denitrification treatment of waste liquid.
Preferably, the waste liquid is high-sodium salt high-concentration nitrate waste liquid.
Preferably, the Pannonibacter phragmittus has a tolerance sodium chloride salt concentration of 1-10% and tolerance nitrate ion (NO)3 -) The concentration was 2000-75000 mg/L.
Preferably, the waste stream contains BOD/NO3 -Is (0.6-2.5): 1.
more preferably, BOD/NO in said waste liquid3 -Is (1-2.5): 1.
preferably, the waste liquid treatment temperature is 18-40 ℃, and the pH is 4.0-8.0.
More preferably, the waste liquid treatment temperature is 20-35 ℃, and the pH is 6.0-7.5.
Preferably, the method for applying the Pannonibacter phragmitetus in the denitrification and denitrification treatment of the waste liquid is as follows:
fermenting Pannonibacter phragmitetus, diluting to obtain bacterium with 10 bacterial activity3-104cfu/L bacterial suspension, mixing the bacterial suspension with sludge according to the volume ratio of 5-15:100, and adding the mixture into waste liquid as an inoculum; the hydraulic retention time of the waste liquid is 12-48 h.
Wherein the dilution method of the dry thallus powder is the same as the preparation method of the bacterial suspension, namely, the dry thallus powder is diluted by utilizing a fermentation culture medium or saline containing 30-60g/L of sodium chloride to obtain the thallus vitality of 103-104cfu/L bacterial suspension.
More preferably, the Pannonibacter phragmitetus is applied to the denitrification and denitrification treatment of the waste liquid by the following specific steps:
the Pannonibacter phragmitetus is diluted into the strain with the bacterial activity of 10 after being fermented3-104The method comprises the steps of mixing cfu/L bacterial suspension with activated sludge according to a volume ratio of 5-15:100, adding the bacterial suspension into a waste liquid reactor, carrying out mixed reaction for 16-24 hours, injecting waste liquid into the waste liquid reactor, and enabling the hydraulic retention time of the waste liquid to be 12-48 hours. The obtained denitrogenation rate is more than 99%.
More preferably, the Pannonibacter phragmitetus is applied to the denitrification and denitrification treatment of the waste liquid by a specific method comprising the following steps:
the waste liquid adopts a two-stage anaerobic treatment method, firstly, anaerobic acidification flora is utilized to carry out acidification treatment on the waste liquid, organic matters which are difficult to degrade are rapidly subjected to ring-opening chain breaking to convert the organic matters into VFA with better biodegradability, the anaerobic acidification reaction time is controlled to be 4-8h, the internal circulation ratio is controlled to be 100-. The obtained denitrogenation rate is more than 99%. The Pannonibacter phragmitetus thallus and sludge mixture is obtained by mixing and reacting for 16-24 hours according to the preparation method.
The internal circulation ratio is the ratio of the flow of the backflow waste liquid to the flow of the inflow waste liquid.
It is noted that the sludge is conventional excess sludge obtained by a common wastewater biochemical treatment system.
Has the advantages that:
compared with the prior art, the Pannonibacter phragmitetus and the application thereof have the following advantages:
(1) the Pannonibacter phragmiteus disclosed by the invention has strong tolerance to high salt, can realize denitrification under the condition of high salt, solves the problem of limitation of high salt on conventional biological treatment, and enables high-concentration nitrate waste liquid to have an economic, efficient, stable and thorough treatment method.
The Pannonibacter phragmitetus has high salt-tolerant denitrification property, the denitrification rate reaches over 99 percent, and high-concentration nitrate Nitrogen (NO) in water can be obtained under the condition that the salt content is less than or equal to 8 percent3 -Less than or equal to 75000mg/L) into harmless nitrogen and no nitrite accumulation, and can be used in a denitrification reactor for efficiently and stably denitrifying without secondary pollution and shortening the startup period.
(2) The method can complete the synchronous removal of carbon and nitrogen, and has better economic benefit and environmental protection benefit for removing nitrate from the high-concentration organic waste liquid.
(3) The strain can utilize various carbon sources and has good adaptability.
In the invention, the Pannonibacter phragmitetus can effectively utilize nitrate in the waste liquid as a nitrogen source to decompose efficiently.
The Pannonibacter phragmittus can effectively utilize a fermentation product VFA (volatile fatty acid) and the like produced after fermentation and acidification by anaerobic acidification flora as an electron donor to carry out denitrification reaction, thereby achieving the effects of denitrification and removal of organic carbon in organic waste liquid and waste liquid.
(4) The microbial inoculum prepared by the culture medium through fermentation culture is added into an anaerobic denitrification reactor, so that the acclimatization starting efficiency of treating the high-salt nitrate waste liquid can be accelerated.
(5) The Pannonibacter phragmitetus provided by the invention can take an organic matter as an electron donor and NO under the condition of high salt3 -It is an electron acceptor, reduced to nitrogen. The strain is used for treating waste liquid, and has the advantages of simple process, thorough denitrification, stable effect and operation cost saving. In practical application, the bacterial strain can be placed in high-salt high-nitrate waste liquid to realize the aim of denitrification.
(6) And (3) denitrification parameter setting: please note that BOD/NO in the present invention3 -The proportion setting effectively ensures the organic electron donor required by the complete denitrification of the microbial inoculum and effectively controls BOD/NO3 ->0.6, the denitrification efficiency is prevented from being reduced. Secondly, the temperature and pH parameter control in the invention can effectively ensure the normal operation of the metabolic reaction of the bacterial strain and maintain the obvious microbial activity, and when the temperature and pH parameter control exceeds the range, the functional microbial activity can be inhibited, and the treatment effect can be deteriorated. The preferred scheme of the invention adds the mixed sludge of the microbial inoculum into the wastewater, and the pure bacteria are attached to the sludge and grow, so that on one hand, the loss of the pure bacteria from the reactor is avoided, on the other hand, the pure bacteria in the reactor are started successfully to become dominant bacteria, and the structures of other microbial populations in the sludge are evolved to form a cooperative metabolic symbiotic relationship with the pure bacteria, thereby enhancing the metabolic rate of functional microorganisms. In addition, the activated sludge can serve as a carrier for the attachment and growth of pure bacteria on one hand, and on the other hand, the actual wastewater pollutant components are complex, if the denitrification effect which can inhibit Pannonibacter phragmitetus is not removed, the activated sludge strains are rich, other pollutants can be removed through domestication, and the inhibition effect of other pollutants on functional bacteria is reduced. The control of the mixing time of the microbial inoculum and the activated sludge effectively ensures the effective attached growth of the strains on the sludge; in addition, the hydraulic retention time in the invention has a remarkable improvement effect on the denitrification rate.
Drawings
FIG. 1Pannonibacter phragmitetus phylogenetic tree;
FIG. 2 Transmission Electron micrograph of Pannonibacter phragmitetus.
Detailed Description
The invention is described below by means of specific embodiments. In addition, the embodiments should be considered illustrative, and not restrictive, of the scope of the invention, which is defined solely by the claims. It will be apparent to those skilled in the art that various changes or modifications in the components and amounts of the materials used in these embodiments can be made without departing from the spirit and scope of the invention. The present invention will be further described with reference to the following examples, but the present invention is not limited to the following examples.
Example 1: isolation and identification of Pannonibacter phragmitetus
(1) Enrichment, isolation medium and propagation medium:
enrichment culture medium: potassium nitrate 2g, magnesium sulfate heptahydrate 0.2g, dipotassium hydrogen phosphate 1.76g, potassium sodium tartrate 20g, 7-hydrate ferrous sulfate 0.005g, calcium chloride 0.02g, ammonium chloride 0.63g, NaCl 5%, distilled water 1000mL, pH 7.5.
Separating a culture medium: potassium nitrate 2g, magnesium sulfate heptahydrate 0.2g, dipotassium hydrogen phosphate 0.5g, sodium potassium tartrate 20g, NaCl 5%, distilled water 1000mL, pH 7.0-7.2, agar 1.5% and 1mL of bromothymol blue solution as solid medium.
Fermentation medium: 8g of peptone, 4g of yeast extract, 10g of sodium chloride and 1000mL of distilled water, wherein the pH value is 7.5.
(2) Isolation and purification of Pannonibacter phragmitetus strain:
weighing 5g of domesticated denitrification sludge, placing the domesticated denitrification sludge in a 250mL anaerobic bottle, adding 100mL of sterilization enrichment culture medium, blowing nitrogen to discharge oxygen, sealing, placing the domesticated denitrification sludge in a 30 ℃ constant temperature incubator for culturing for 4 days, then taking 2mL of mixed bacteria liquid, inoculating the mixed bacteria liquid in an anaerobic glove box to a selective culture medium, and repeating the operation for 3 times. Then 1mL of the suspension was aspirated from the selection medium to 9mL of a dilution (sterile, oxygen-free water) to give 10-2Diluting the suspension by 10 timesDiluting to 10-7Thus, each dilution of bacterial suspension was prepared.
0.1mL of each dilution suspension is respectively sucked in an anaerobic glove box and evenly coated on the denitrifying bacteria separation solid culture medium. Then, the plate is inverted and placed in a constant temperature incubator at 30 ℃ to be cultured until obvious bacterial colonies grow. Then, a single strain was picked up and the Pannonibacter phragmitetus of the present invention was purified by streaking multiple times on a plate.
(3) PCR amplification and sequencing of 16S rRNA:
the DNA of Pannonibacter phragmitetus is extracted and purified by a Smart LabAssist-16 and MO-BIO PowerSoil DNA extraction kit (Taiwan dot nanotechnology Co., Ltd.) of a full-automatic magnetic bead nucleic acid extractor. Taking 5 mu L of DNA sample, carrying out electrophoresis detection under the conditions of 120V and 20-25min, and observing quality under a gel electrophoresis apparatus.
Bacterial universal primers F16S-27(5 '-AGAGTTTGATCCTGGCTCAG-3') and R16S-1492(5 '-CGGTTACCTTGTTACGACTTC-3') are selected for PCR amplification. The PCR reaction conditions are as follows: pre-denaturation at 94 ℃ for 3min, denaturation at 94 ℃ for 30s, annealing at 54 ℃ for 30s, extension at 72 ℃ for 90s, and circulation for 30 cycles; then extending for 10min at 72 ℃; finally, the mixture is stored at 4 ℃.
The PCR amplification product after being recovered is ordered to sequence by Beijing Olympic Gene science and technology Limited.
And (3) obtaining a sequence with the length of the 16S rDNA of the strain being 1430bp after sequencing, comparing the sequence with other strains when the sequence is submitted to Genbank, constructing a phylogenetic tree by using a phylogenetic analysis (MEGA3.1) software ortho-position connection (Neighbour Joining) method to find that the strain is the closest to the evolution distance of Pannonibacter sp, and determining that the strain belongs to the Pannonibacter genus and is named Pannonibacter phragmitetus.
(4) Denitrification condition testing of Pannonibacter phragmitetus
100mL of sterilized enrichment medium with the nitrate concentration of 2000mg/L and the salt content of 3% is filled into a 250mL triangular flask, inoculated into seed bacterial liquid according to the inoculation amount of 10%, and subjected to standing culture. The Pannonibacter phragmitetus can remove more than 85 percent of total nitrogen within three days at the temperature of 25-35 ℃, and the total nitrogen removal rate can reach as high as 99 percent. The nitrite accumulation phenomenon occurs in the first 48 hours of the culture of the strain, then the nitrite concentration is gradually reduced, and the total nitrogen is reduced to be below 20 mg/L.
Example 2: preparation of Pannonibacter phragmitetus microbial inoculum
After activated culture, Pannonibacter phragmititus is inoculated into a fermentation medium according to the inoculation amount of 5 percent (volume ratio), and the fermentation culture conditions are controlled as follows: 35 ℃ dissolved oxygen<0.2mg/L, pot pressure: shaking and culturing at 40-45rpm under 0.03-0.10Mpa for 72h to obtain fermentation liquid; centrifuging the fermentation liquid at 4 deg.C and 7500rpm for 10min, removing supernatant, diluting the precipitate with sterile fermentation medium to obtain 10-strain active substance3-104The bacterial suspension is filled to form a liquid Pannonibacter phragmitetus microbial inoculum product which is used for denitrifying and denitrifying high-salt high-concentration nitrate waste liquid.
The fermentation medium comprises the following components: 10g/L of peptone, 5g/L of yeast extract, 50g/L of sodium chloride and the balance of distilled water, wherein the pH value is 7.5, the sterilization is carried out at 121 ℃ for 20min, and the sterilization is carried out and then the cooling is carried out to the room temperature.
Example 3: preparation of Pannonibacter phragmitetus microbial inoculum
And (3) inoculating the Pannonibacter phragmitetus into a seed culture medium, controlling the culture temperature to be 35 ℃, and carrying out shake culture at 40rpm for 48h to obtain seed fermentation liquor.
Inoculating the seed fermentation liquid into a fermentation culture medium according to the inoculation amount of 10% (volume ratio), and controlling the fermentation culture conditions as follows: 35 ℃, dissolved oxygen <0.2mg/L, pot pressure: 0.03-0.10Mpa, and shaking culture at 30-50rpm for 72 h.
The preparation method of the microbial inoculum comprises the following steps: fermenting the Pannonibacter phragmitetus, centrifuging at 4 ℃ and 7500rpm for 10min, removing supernatant to obtain thallus precipitate, diluting the thallus precipitate with fermentation medium to obtain thallus vitality of 103And (3) cfu/L bacterial suspension is filled to finally obtain the Pannonibacter phragmitetus microbial inoculum.
The seed liquid culture medium comprises the following components: KNO3 3g/L、MgSO4·7H2O 0.3g/L、K2HPO40.6g/L, 20g/L of potassium sodium tartrate, 50g/L of sodium chloride and the balance of water, the pH value is 7.2, and the sterilization is carried out for 20min at the temperature of 121 ℃.
The fermentation medium comprises the following components: 8g/L of peptone, 4g/L of yeast extract, 50g/L of sodium chloride and the balance of distilled water, wherein the pH value is 7.5, the sterilization is carried out at 121 ℃ for 20min, and the sterilization is carried out and then the cooling is carried out to the room temperature.
Example 4: preparation of Pannonibacter phragmitetus microbial inoculum
And (3) inoculating the Pannonibacter phragmitetus into a seed culture medium, controlling the culture temperature to be 30 ℃, and performing shake culture at 45rpm for 50h to obtain seed fermentation liquor.
Inoculating the seed fermentation liquid into a fermentation culture medium according to the inoculation amount of 5% (volume ratio), and controlling the fermentation culture conditions as follows: 30 ℃, dissolved oxygen <0.2mg/L, pot pressure: 0.06Mpa, 40rpm shake culture time 72 h.
The preparation method of the microbial inoculum comprises the following steps: fermenting the Pannonibacter phragmitetus, centrifuging at the temperature of 4 ℃ and the rpm of 6500 for 10min, removing supernatant to obtain thallus precipitate, diluting the thallus precipitate with a fermentation culture medium to obtain the thallus vitality of 104And (3) cfu/L bacterial suspension is filled to finally obtain the Pannonibacter phragmitetus microbial inoculum.
The seed liquid culture medium comprises the following components: KNO3 3g/L、MgSO4·7H2O 0.3g/L、K2HPO40.6g/L, 25g/L of potassium sodium tartrate, 30g/L of sodium chloride and the balance of water, the pH value is 7.2, and the sterilization is carried out for 20min at the temperature of 121 ℃.
The fermentation medium comprises the following components: 8g/L of peptone, 4g/L of yeast extract, 40g/L of sodium chloride and the balance of distilled water, wherein the pH value is 7.5, the sterilization is carried out at 121 ℃ for 20min, and the sterilization is carried out and then the cooling is carried out to the room temperature.
Example 5: preparation of Pannonibacter phragmitetus microbial inoculum
And (3) inoculating the Pannonibacter phragmitetus into a seed culture medium, controlling the culture temperature to be 40 ℃, and carrying out shake culture at 50rpm for 48h to obtain seed fermentation liquor.
Inoculating the seed fermentation liquor into a fermentation culture medium according to the inoculation amount of 3 percent (volume ratio), and controlling the fermentation culture conditions as follows: 20 ℃, dissolved oxygen <0.2mg/L, tank pressure: 0.03Mpa, and shaking culture at 35rpm for 72 h.
The preparation method of the microbial inoculum comprises the following steps: fermenting the Pannonibacter phragmitetus, centrifuging at 5 ℃ and 6000rpm for 10min, removing supernatant to obtain thallus precipitate, diluting the thallus precipitate with a fermentation culture medium to obtain thallus activity of 9 × 103And (4) filling the bacterial suspension to finally obtain the Pannonibacter phragmitetus microbial inoculum.
The seed liquid culture medium comprises the following components: KNO3 2g/L、MgSO4·7H2O 0.2g/L、K2HPO40.5g/L, 20g/L of potassium sodium tartrate, 60g/L of sodium chloride and the balance of water, the pH value is 7.2, and the sterilization is carried out for 20min at the temperature of 121 ℃.
The fermentation medium comprises the following components: 6g/L of peptone, 3g/L of yeast extract, 60g/L of sodium chloride and the balance of distilled water, wherein the pH value is 7.5, the sterilization is carried out at 121 ℃ for 20min, and the sterilization is carried out and then the cooling is carried out to the room temperature.
Example 6: preparation of Pannonibacter phragmitetus microbial inoculum
And (3) inoculating the Pannonibacter phragmititus into a seed culture medium, controlling the culture temperature to be 38 ℃, and shaking the culture at 55rpm for 72h to obtain seed fermentation liquor.
Inoculating the seed fermentation liquid into a fermentation culture medium according to the inoculation amount of 10% (volume ratio), and controlling the fermentation culture conditions as follows: 40 ℃, dissolved oxygen <0.2mg/L, pot pressure: 0.10Mpa, and shaking culture time of 55rpm for 96 h.
The preparation method of the microbial inoculum comprises the following steps: fermenting the Pannonibacter phragmitetus, centrifuging at 10 ℃ and 7500rpm for 15min, discarding the supernatant to obtain a thallus precipitate, diluting the thallus precipitate with a fermentation medium to obtain the thallus vitality of 8 × 103The Pannonibacter phragmitetus microbial inoculum is finally obtained after filling.
The seed liquid culture medium comprises the following components: KNO3 4g/L、MgSO4·7H2O 0.4g/L、K2HPO40.8g/L, 30g/L of sodium potassium tartrate, 45g/L of sodium chloride and the balance of water, the pH value is 7.2, and the sterilization is carried out for 20min at the temperature of 121 ℃.
The fermentation medium comprises the following components: peptone 12g/L, yeast extract 6g/L, sodium chloride 45g/L, and distilled water in balance, with pH 7.5, sterilizing at 121 deg.C for 20min, sterilizing, and cooling to room temperature.
Example 7: preparation of Pannonibacter phragmitetus microbial inoculum
And (3) inoculating the Pannonibacter phragmitetus into a seed culture medium, controlling the culture temperature to be 35 ℃, and carrying out shake culture at 35rpm for 60 hours to obtain seed fermentation liquor.
Inoculating the seed fermentation liquid into a fermentation culture medium according to the inoculation amount of 5% (volume ratio), and controlling the fermentation culture conditions as follows: 35 ℃, dissolved oxygen <0.2mg/L, pot pressure: 0.05Mpa, and the shaking culture time at 35rpm is 75 h.
The preparation method of the microbial inoculum comprises the following steps: the Pannonibacter phragmiteus is fermented, centrifuged at 6000rpm at 4 ℃ for 15min, the supernatant is discarded to obtain thallus precipitate, lyophilized to obtain microbial inoculum dry powder, and the microbial inoculum dry powder is filled to finally obtain the Pannonibacter phragmiteus microbial inoculum.
The seed liquid culture medium comprises the following components: the same as in example 3;
the fermentation medium comprises the following components: the same as in example 3.
Example 8: pannonibacter phragmitetus for treating high-salt high-nitrate waste liquid
The microbial inoculum product prepared in the embodiment 3 of the invention is mixed with the sludge in the conventional denitrification reactor according to the volume ratio of 1:10, stopping feeding the waste liquid, avoiding the loss of the microbial inoculum, ensuring the retention time of the microbial inoculum in the reactor to be 20 hours, and controlling the BOD/NO of the waste liquid feeding water of the reactor3 -(0.8-1.5): 1, controlling the reaction temperature at 30-35 ℃, controlling the pH at 6.5-7.5, gradually increasing the concentrations of salt and nitrate radical of inlet water in equal steps, finally increasing the concentration of sodium chloride salt from 1% to 8% and the concentration of nitrate radical from 2000mg/L to 50000mg/L in inlet water, and ensuring that the hydraulic retention time of waste liquid is 36 hours and NO NO exists2 -Accumulating, and after treatment, the nitrate nitrogen in the effluent is reduced to 20mg/L and NO3 -The concentration is kept below 80 mg/L.
Example 9: pannonibacter phragmitetus for treating high-salt high-nitrate waste liquid
The microbial inoculum product prepared in the embodiment 4 of the invention is mixed with the sludge in the conventional denitrification reactor according to the volume ratio of 1:10, stopping feeding the waste liquid, avoiding the loss of the microbial inoculum, ensuring the retention time of the microbial inoculum in the reactor to be 24 hours, and then controlling the BOD/NO of the waste liquid feeding water of the reactor3 -0.8-1: 1, controlling the reaction temperature at 20-30 ℃, controlling the pH at 4.5-5.5, gradually increasing the salt concentration and the nitrate concentration of inlet water in equal steps, finally increasing the sodium chloride salt concentration from 1.5% to 10%, increasing the nitrate ion concentration from 2000mg/L to 75000mg/L, and keeping the hydraulic retention time for 48 hours without NO2 -Accumulating, after treatment, the nitrate nitrogen in the effluent waste liquid is reduced to below 50mg/L, and the effluent NO3 -The concentration is kept below 150 mg/L.
Example 10: pannonibacter phragmitetus for treating high-salt high-nitrate waste liquid
The microbial inoculum product prepared in the embodiment 5 of the invention is mixed with the sludge in the conventional denitrification reactor according to the volume ratio of 15:100 is mixed and added into a waste liquid reactor, the waste liquid feeding is stopped, the loss of the microbial inoculum is avoided, the retention time of the microbial inoculum in the reactor is ensured to be 16 hours, and then the BOD/NO of waste liquid feeding water of the reactor is controlled3 -Controlling the reaction temperature to be between 30 and 40 ℃ and the pH to be between 5.5 and 6.5, gradually increasing the salt concentration and the nitrate concentration of inlet water in equal steps, finally increasing the sodium chloride salt concentration from 1 percent to 10 percent, increasing the nitrate ion concentration from 10000mg/L to 60000mg/L, and keeping the hydraulic retention time for 48 hours without NO2 -And (4) accumulating. After treatment, nitrate nitrogen in the waste liquid is reduced to below 25mg/L and NO is discharged3 -The concentration is kept below 100 mg/L.
Example 11: pannonibacter phragmitetus for treating high-salt high-nitrate waste liquid
The microbial inoculum product prepared in the embodiment 6 of the invention is mixed with the sludge in the conventional denitrification reactor according to the volume ratio of 1: 20 is mixed and added into a waste liquid reactor, the waste liquid feeding is stopped, and theThe loss of the microbial inoculum is avoided, the retention time of the microbial inoculum in the reactor is ensured to be 16 hours, and then the BOD/NO of wastewater inlet water of the reactor is controlled3 -1.5:1, controlling the reaction temperature at 30-35 ℃, controlling the pH value at 7.5-8.0, gradually increasing the salt concentration and the nitrate concentration of inlet water in equal steps, finally increasing the sodium chloride salt concentration from 1.5% to 10% and the nitrate ion concentration from 4000mg/L to 75000mg/L in the inlet water, and keeping the hydraulic retention time for 45 hours without NO2 -And (4) accumulating. After treatment, the nitrate nitrogen in the waste liquid is reduced to below 50mg/L and NO is discharged3 -The concentration is kept below 150 mg/L.
Example 12: pannonibacter phragmitetus for treating high-salt high-nitrate waste liquid
The microbial inoculum dry powder prepared in the embodiment 7 of the invention is diluted by an aseptic fermentation medium until the bacterial activity is 104The bacterial suspension is mixed with sludge according to the volume ratio of 15:100 and then is added into a reactor, water feeding is stopped to avoid loss of the bacterial agent, the retention time of the bacterial agent in the reactor is ensured to be 16 hours, then waste liquid is fed, and BOD/NO of waste liquid water fed into the reactor is controlled3 -1, controlling the reaction temperature at 30-35 ℃, controlling the pH value at 7.5-8.0, gradually increasing the concentrations of the salt and the nitrate radical of the inlet water in equal steps, finally increasing the concentration of the sodium chloride salt in the inlet water from 1% to 10%, increasing the concentration of the nitrate radical ion in the inlet water from 3000mg/L to 75000mg/L, and keeping the hydraulic retention time for 36 hours without NO2 -And (4) accumulating. After treatment, the nitrate nitrogen in the waste liquid is reduced to below 50mg/L and NO is discharged3 -The concentration is kept below 150 mg/L.
Example 13: pannonibacter phragmitetus for treating high-salt high-nitrate waste liquid
Diluting Pannonibacter phragmitetus thallus with sterile fermentation medium until the activity is 104The bacterial suspension is prepared by mixing a bacterial dilution liquid and sludge according to the volume ratio of 1:10, adding the mixture into a waste liquid reactor, stopping water inflow to avoid the loss of the bacterial agent, and ensuring that the retention time of the bacterial agent in the reactor is 15 hours.
The waste liquid is treated with two-stage anaerobic treatment process, including the first anaerobic acidifying floraAcidifying, quickly opening the ring and breaking the chain of organic matters difficult to degrade to convert the organic matters into VFA with better biodegradability, controlling the anaerobic acidification reaction time to be 4-8h and the internal circulation ratio to be 100-3 -The reaction temperature, pH, feed water salt and nitrate maximum concentration, and hydraulic retention time of the waste liquid were the same as in example 12.
The Pannonibacter phragmittetus uses nitrate nitrogen as an electron acceptor and VFA generated by anaerobic acidification as an electron donor to carry out anaerobic denitrification reaction to treat sewage, and the internal circulation ratio is controlled to be 100-300 percent.
After treatment, the nitrate nitrogen in the waste liquid is reduced to below 50mg/L and NO is discharged3 -The concentration is kept below 150 mg/L.
Comparative example 1 treatment of high-salt high-nitrate waste liquid with conventional sludge
The liquid waste treatment was carried out according to the treatment method of example 8, the same amount of sludge was finally added to the reactor, and the operation was carried out according to the procedure of example 8, which is different from example 8 in that: the sludge is not externally inoculated with a microbial inoculum, and the difference is that in the final inlet water, the concentration of sodium chloride salt is improved from 0.5 percent to 3 percent, the concentration of nitrate ion is improved from 500mg/L to 8000mg/L, the hydraulic retention time of the waste liquid is 36 hours, and the NO of the outlet water3 -At a concentration of 60mg/L and NO2 -Significant accumulation of NO2 -The concentration was 550mg/L and the ORP increased to +110mv, resulting in a continuous deterioration of the treatment effect. According to the experimental effect data, the waste liquid which can continuously improve the concentration of sodium chloride salt and the concentration of nitrate ions through conventional sludge treatment can not achieve good denitrification effect.
Sequence listing
<110> Dajiang environmental shares Ltd
<120> denitrifying bacterium tolerant to high salt and capable of efficiently degrading nitrate, and preparation and application thereof
<130> 1
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1353
<212> DNA
<213> Pannonibacter phragmitetus CGMCC No.15312
<400> 1
catgcagtcg aacgcatcgc agatgagtgg cagacgggtg agtaacacgt gggaacatac 60
cctttggtgc ggaacaacag ctggaaacgg ctgctaatac cgcatgcgcc ctacggggga 120
aagatttatc gccgaaggat tggcccgcgt ctgattagct agttggtgag gtaatggctc 180
accaaggcga cgatcagtag ctggtctgag aggatgatca gccacactgg gactgagaca 240
cggcccagac tcctacggga ggcagcagtg gggaatattg gacaatgggc gcaagcctga 300
tccagccatg ccgcgtgagt gatgaaggcc ctagggttgt aaagctcttt cagcgaggag 360
gataatgacg ttactcgcag aagaagcccc ggctaacttc gtgccagcag ccgcggtaat 420
acgaaggggg ctagcgttgt tcggaatcac tgggcgtaaa gcgcacgtag gcggactttt 480
aagtcagggg tgaaatcccg gggctcaacc tcggaactgc ctttgatact ggaagtctgg 540
agtccgagag aggtgagtgg aactccgagt gtagaggtga aattcgtaga tattcggaag 600
aacaccagtg gcgaaggcgg ctcactggct cggtactgac gctgaggtgc gaaagcgtgg 660
ggagcaaaca ggattagata ccctggtagt ccacgccgta aacgatggaa gctagccgtc 720
agggtgcatg catcttggtg gcgcagttaa cgcattaagc ttcccgcctg gggagtacgg 780
tcgcaagatt aaaactcaaa ggaattgacg ggggcccgca caagcggtgg agcatgtggt 840
ttaattcgaa gcaacgcgca gaaccttacc agctcttgac atttggtgct accacgggag 900
accgtgggtt cccttcgggg acgccaggac aggtgctgca tggctgtcgt cagctcgtgt 960
cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct cgcccttagt tgccagcatt 1020
gagttgggca ctctaggggg actgccggtg ataagccgag aggaaggtgg ggatgacgtc 1080
aagtcctcat ggcccttacg ggctgggcta cacacgtgct acaatggcgg tgacagtggg 1140
cagcgaaggg gtgacccgga gctaatctcc aaaagccgtc tcagttcgga ttgttctctg 1200
caactcgaga gcatgaagtt ggaatcgcta gtaatcgcgt aacagcatga cgcggtgaat 1260
acgttcccgg gccttgtaca caccgcccgt cacaccatgg gagttgggtt tacccgaagg 1320
tggtgcgcta accgcaagga ggcagccaac cac 1353

Claims (7)

1. A denitrifying bacterium which can tolerate high-salt and efficiently degrade nitrate is Pannonibacter phragmitetus, and the preservation number is CGMCC No. 15312.
2. The microbial inoculum containing the high-salt-tolerance nitrate-degrading denitrifying bacteria of claim 1, which is characterized in that: the microbial inoculum is specifically dry microbial inoculum powder or the vitality of the microbial inoculum is 103-104cfu/L bacterial suspension.
3. The method for preparing the high-salt-tolerance denitrifying bacteria microbial inoculum for efficiently degrading nitrate according to claim 2, which is characterized in that:
(1) fermentation culture: activating the Pannonibacter phragmititus, inoculating into a fermentation culture medium according to the inoculation amount of 3% -10%, and controlling the fermentation culture conditions as follows: 20-40 ℃, dissolved oxygen <0.2mg/L, pot pressure: shaking and culturing at 0.02-0.10Mpa at 30-55rpm for 70-96 h; the fermentation medium comprises the following components: 6g/L-12g/L peptone, 3g/L-6g/L yeast extract, 30-60g/L sodium chloride, and the balance of distilled water, wherein the pH value is 7.5, and the sterilization is carried out at 121 ℃ for 20 min;
(2) and (2) centrifuging and freeze-drying the fermentation liquor obtained in the step (1) to obtain thallus dry powder or centrifuging and diluting the thallus to obtain a thallus suspension.
4. The preparation method of the high-salt-tolerance denitrifying bacteria microbial inoculum for efficiently degrading nitrate according to claim 3, which is characterized by comprising the following steps:
the activation steps of the Pannonibacter phragmitetus are as follows: inoculating the Pannonibacter phragmitetus into a seed culture medium, controlling the culture temperature to be 30-40 ℃, and shaking the culture time to be 45-72h at 30-55 rpm; the seed liquid culture medium comprises the following components: KNO3 2g/L-5g/L、MgSO4·7H2O 0.2g/L-0.5g/L、K2HPO40.5-0.8 g/L, 20-30 g/L of sodium potassium tartrate, 30-60g/L of sodium chloride and the balance of water, the pH value is 7.2, and the sterilization is carried out for 20min at 121 ℃.
5. The use of the high-salt-tolerance, nitrate-degrading bacteria of claim 1 in denitrification of waste liquid.
6. The use of the high-salt-tolerance and high-efficiency nitrate-degrading denitrifying bacteria according to claim 5 in the denitrification treatment of waste liquid, which is characterized in that: the concentration of sodium chloride salt in the waste liquid is 1-10%, the concentration of nitrate ions is 2000-75000mg/L, BOD/NO3 -Is (0.6-2.5): 1; in the waste liquid, the treatment temperature of the waste liquid is 18-40 ℃, and the pH value is 4.0-8.0.
7. The use of the denitrifying bacteria that are tolerant to high salt and efficiently degrade nitrate according to claim 6 in the denitrification and denitrification treatment of waste liquid, wherein: the Pannonibacter phragmitetus is diluted into the strain with the bacterial activity of 10 after being fermented3-104cfu/L bacterial suspension, mixing the bacterial suspension with sludge in a volume ratio of 5-15:100 to serve as an inoculum, and adding the inoculum into waste liquid; the hydraulic retention time of the waste liquid is 12-48 h.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101602060A (en) * 2009-07-03 2009-12-16 中南大学 A kind of repairing method of microorganism in contaminated soil of chromium slag storage yard
CN101684453A (en) * 2009-07-03 2010-03-31 中南大学 Bacterial strain for repairing chromium slag contaminated site
CN102108334A (en) * 2009-12-25 2011-06-29 中国科学院过程工程研究所 Chromium-removing rhodobacterium and application thereof
CN102127517A (en) * 2010-12-20 2011-07-20 中南大学 Strain with heavy metal tolerance and applications thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101602060A (en) * 2009-07-03 2009-12-16 中南大学 A kind of repairing method of microorganism in contaminated soil of chromium slag storage yard
CN101684453A (en) * 2009-07-03 2010-03-31 中南大学 Bacterial strain for repairing chromium slag contaminated site
CN102108334A (en) * 2009-12-25 2011-06-29 中国科学院过程工程研究所 Chromium-removing rhodobacterium and application thereof
CN102127517A (en) * 2010-12-20 2011-07-20 中南大学 Strain with heavy metal tolerance and applications thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
High-efficiency inorganic nitrogen removal by newly isolated Pannonibacter phragmitetus B1;Hong Bai et al.;《Bioresource Technology》;20180918;第271卷;第91-99页 *

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