CN110117567B - Paracoccus denitrificans strain screening and application thereof in deodorization - Google Patents
Paracoccus denitrificans strain screening and application thereof in deodorization Download PDFInfo
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Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to paracoccus denitrificans, which has the following strain names: paracoccus denitrificans, paracoccus Denitrificans TS-1; the preservation number is: CGMCC NO.17605. The paracoccus denitrificans has a 16SrDNA sequence shown as SEQ ID No. 1. Meanwhile, the invention also discloses application of paracoccus denitrificans TS-1 in deodorization and sewage treatment. The paracoccus denitrificans TS-1 has high heterotrophic nitrification and aerobic denitrification capabilities and high sulfide removal capability, and is a strain with three functions.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to paracoccus denitrificans strain screening and application thereof in deodorization.
Background
The malodorous pollutants refer to all gas substances which stimulate olfactory senses to cause people to be unpleasant and harm living environment, the substances are various, the influence range is large, and the treatment is gradually paid attention to by people. A large amount of deodorizing microorganisms exist in the livestock manure and domestic garbage, and a large amount of ammonia gas and hydrogen sulfide are generated in the garbage landfill and composting process, so that the livestock manure not only causes serious harm to human bodies, but also pollutes the environment, underground water sources and the like.
The current conventional deodorization methods are: physical deodorization, chemical deodorization, biological deodorization. The physical deodorization method eliminates the odor by means of conversion among solid, liquid and gas phases, only reduces the perception degree of smell by smell, but does not change the chemical properties of the odor fundamentally. Chemical deodorization is the addition of certain chemical agents that change their chemical structure to destroy their deodorizing groups, turning them into odorless or less odorous substances. The biological method is a novel malodor treatment method developed in recent years, and a mainstream method of deodorization is being developed gradually because of advantages such as low investment and running costs, high treatment efficiency, no secondary pollution, and the like.
The biological method comprises a biological washing method, a biological filter method and a biological trickling filtration method. Compared with other two methods, the biological trickling filter can accurately control reaction conditions (such as humidity, pH and the like), and obviously improves the removal efficiency. In the deodorization process of compost plants, refuse landfills and farms, the high-efficiency microbial inoculum is connected into the deodorization reactor, so that odor can be quickly removed, no odor or secondary pollution is realized, the better selection of environmental purification such as compost deodorization and refuse landfills is realized, and the wide popularization and application can be realized.
Disclosure of Invention
The invention aims to provide paracoccus denitrificans capable of efficiently denitrifying and desulfurizing, and the paracoccus denitrificans has good capability of degrading ammonia nitrogen, nitrate nitrogen and sulfide. The method can be used for degrading and treating wastewater containing ammonia nitrogen, nitrate nitrogen and inorganic sulfides, and can be applied to a biological trickling filter reactor for treating toxic and harmful gases such as ammonia gas, hydrogen sulfide and the like.
In order to realize the purpose of the invention, the technical scheme adopted by the invention is as follows:
a paracoccus denitrificans, the strain name: paracoccus denitrificans, paracoccus dentiricans TS-1, accession number: CGMCC NO.17605.
Preferably, the 16SrDNA sequence of the paracoccus denitrificans TS-1 is shown as SEQ ID No. 1.
Preferably, the paracoccus denitrifican is used for deodorization.
Preferably, the application of paracoccus denitrifican in deodorization comprises the following specific steps:
(1) Culturing paracoccus denitrificans TS-1 into paracoccus denitrificans TS-1 bacterial liquid, wherein the active bacterial amount of the paracoccus denitrificans TS-1 in the bacterial liquid is 10 8 ～10 9 CFU/mL；
(2) Adding a solution which provides nutrition required by growth of paracoccus denitrificans TS-1 into a biological trickling filter filled with a filler, and then adding the bacterial liquid of the paracoccus denitrificans TS-1 obtained in the step (1);
(3) The air flow rate of 4-6L/min and the gas concentration of 40-60 mg/m 3 The gas inflow and the gas concentration of the hydrogen sulfide are 40-60 mg/m 3 Introducing air, hydrogen sulfide and ammonia gas into the biological trickling filter simultaneously, and performing biofilm culturing and domestication for 6-8 days under the conditions of the ambient temperature of 25-27 ℃, the pH value of 6.5-7.5 and the humidity of 38% -45%;
(4) And introducing gas to be treated into the biological trickling filter after the membrane hanging acclimation is finished.
Preferably, in the step (1), the paracoccus denitrificans TS-1 is cultured as a bacterial solution of the paracoccus denitrificans TS-1 under conditions that the culture temperature is 27 to 33 ℃, the pH of the culture medium is 6.0 to 7.0, and the C/N ratio in the culture medium is 12 to 18.
Preferably, in the step (1), the culture of Paracoccus denitrificans TS-1 is performed under conditions of a culture temperature of 30 ℃, a culture medium pH of 6.5 and a C/N ratio of 15 in the culture medium.
Preferably, in the step (4), a solution for providing nutrients required for the growth of paracoccus denitrificans TS-1 is added into the bio-trickling filter while the gas to be treated is introduced.
Preferably, the paracoccus denitrifican is applied to anhydrous treatment.
The invention has the following beneficial effects:
1. the nutrition type of the paracoccus denitrificans TS-1 is facultative nutrition type, the rapid propagation of thalli is realized under the heterotrophic condition, and the efficient desulfurization is realized under the autotrophic condition. The strain is subjected to amplification culture by using an organic carbon source heterotrophic culture medium to obtain high-density thalli, so that ammonia nitrogen can be efficiently degraded in a heterotrophic environment, and sulfide can be oxidized and degraded under an autotrophic condition.
2. The paracoccus denitrificans TS-1 has a rapid ammonia nitrogen degradation rate, can degrade 170mg/L ammonia nitrogen for 10 hours under the optimal condition, and has the degradation rate reaching 17.0 mg/(L & gth).
3. The paracoccus denitrificans TS-1 has a high nitrate nitrogen degradation rate, the bacterial strain can realize a denitrification function under an aerobic condition, and can degrade nitrate nitrogen again to generate nitrogen after ammonia nitrogen is converted into nitrate nitrogen, so that ammonium ions are thoroughly removed. Under the optimal condition, the degradable material can be degraded by about 167mg/L within 10h, namely 16.7 mg/(L.h).
4. The desulfurization efficiency of paracoccus denitrificans TS-1 is high, 100% of sulfide removal rate is achieved within 6 hours under the autotrophic condition, and the maximum conversion rate of elemental sulfur can reach 63.71% in 4.5 hours.
5. The invention accords with the biological safety regulation, and the paracoccus denitrificans TS-1 is screened from the selective environment stressed by high ammonia nitrogen and high sulfide for a long time, so that the damage to the surrounding environment and ecological balance is avoided.
Drawings
FIG. 1 shows the cell morphology of Paracoccus denitrificans TS-1 according to the present invention under a scanning electron microscope.
FIG. 2 is a graph showing the growth of Paracoccus denitrificans TS-1 of the present invention cultured in heterotrophic LB medium.
FIG. 3 is the change of the degradation rate of ammonia nitrogen with time in the experimental process of degrading ammonia nitrogen by Paracoccus denitrificans TS-1 of the invention;
FIG. 4 shows the time-dependent change of the nitrate nitrogen degradation rate of Paracoccus denitrificans TS-1 in the nitrate nitrogen degradation experiment process;
FIG. 5 shows the change of the sulfide removal rate of Paracoccus denitrificans TS-1 with time in the desulfurization test process;
FIG. 6 shows the change of elemental sulfur conversion with time during desulfurization experiments of Paracoccus denitrificans TS-1 according to the present invention;
FIG. 7 shows the elemental sulfur formed by desulfurization of Paracoccus denitrificus TS-1 according to the present invention under a scanning electron microscope;
FIG. 8 phylogenetic tree of Paracoccus denitrificans TS-1.
Detailed Description
The present invention will be described in detail with reference to examples. The embodiments are provided to facilitate a better understanding of the invention and are not intended to limit the invention.
The media referred to in the following examples are:
activated sludge acclimation medium (/ L): KH (Perkin Elmer) 2 PO 4 1.0g，K 2 HPO 4 4.0g，MgCl 2 0.5g，FeSO 4 0.01g, 5.0g of glucose, ammonia nitrogen and sulfide; the addition amounts of ammonia nitrogen and sulfide are shown in the following table:
TABLE-addition of ammonia nitrogen and sulfide in activated sludge acclimation medium (/ L)
| Ⅰ | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |
| Na 2 S | 1.0 | 2.0 | 3.0 | 4.0 | 5.0 |
| (NH4) 2 SO 4 | 0.8 | 1.6 | 2.4 | 3.2 | 4.0 |
Denitrogenation and desulfurization medium I (/ L), glucose 5.0g, naCl 2.0g, KH 2 PO 4 1.0g，MgCl 2 0.5g，FeSO 4 0.01g，(NH4) 2 SO 4 2.0g，Na 2 S1.2 g, the pH was adjusted to 7.0 with 1mol/L HCl solution.
Denitrogenation and desulfurization medium II (/ L): 5g of NaHS.
LB medium (/ L): tryptone 10.0g, yeast extract 5.0g, naCl 10.0g, pH 7.0. The LB medium is used for scale-up culture.
Circulating medium (/ L): glucose 6 g/KH 2 PO 4 1.0g，K 2 HPO 4 1.0g，MgCl 2 0.4g，NaHCO 3 0.4g。
Example 1: strain screening
Collecting 4L of activated sludge of a secondary sedimentation tank of a double-flow sewage treatment plant in Sichuan, adding the activated sludge into a reactor, adding 1L of activated sludge acclimation culture medium to carry out aeration culture, respectively treating each concentration from low concentration to high concentration for 5 days, 7 days and 8 days as a period according to a designed concentration gradient, and carrying out acclimation culture after 30 days of acclimation from low concentration to high concentration. Collecting a small amount of supernatant, enriching in a liquid denitrifying and desulfurizing culture medium for 2 days, repeating for 4-5 times, and performing gradient dilution from 10 times -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 Sucking 0.2mL of the culture medium for flat plate coating in a denitrification and desulfurization medium under dilution, culturing at constant temperature of 30 ℃, obtaining a plurality of bacterial colonies after multiple separations, and selecting the single bacterial colony with the best growth conditionCulturing in denitrogenation and desulfurization culture medium, repeating for more than 5 times to obtain pure bacteria, and naming the bacteria as TS-1. The physiological and biochemical properties of the strain are as follows:
TABLE 2 physiological and biochemical properties of Paracoccus denitrificans CGMCC NO.17605
| Detecting items | The result of the detection | Detecting items | The result of the detection |
| Gram stain | - | V.P | - |
| Shape of cell | Spherical shape | Contact enzyme | + |
| Oxidases | - | M.R | - |
| Starch hydrolysis | + | Gelatin | + |
| Nitrate reduction test | + | Good/anaerobic property | Aerobic |
| Glucose | - | Maltose | - |
| L-arabinose | - | Lactose | - |
| Xylose | - | Grease | + |
Note: "+" indicates positive, "-" indicates negative
Example 2: identification of strains
Inoculating the separated and purified strains into the first denitrogenation and desulfurization culture medium according to the same inoculum size, culturing in shaking tables at different temperatures (5 deg.C, 10 deg.C, 24 deg.C, 27 deg.C, 30 deg.C, 33 deg.C, 36 deg.C, 40 deg.C, 45 deg.C, 50 deg.C) for 12h, and determining OD 600 The growth temperature of the strain is determined to be 10-50 ℃, and the optimal growth temperature is 30 ℃. Culturing at 30 deg.C for 24 hr at different initial pH (3.0, 4.0, 5.0, 6.0, 6.5, 7.0, 7.5, 8.0, 9.0, 10.0, 11.0), and determining OD 600 The growth pH of the strain is determined to be 4.0-10.0, and the optimum growth pH is 6.5. OD was measured at pH 6.5 and temperature 30 ℃ using different C/N (1, 5, 10, 15, 20, 25, 30) 600 The tolerance range of the strain is determined to be 5-25C/N, and the optimal C/N is 15.
The separated and purified strain was observed under a Scanning Electron Microscope (SEM), and the results are shown in fig. 1; the strain is in the shape of short rod, 0.5-0.9 μm long, 0.4-0.7 μm wide and has no flagellum; gram staining identified the bacterium as negative.
A bacterial whole genome rapid extraction kit is adopted to extract the whole genome of a pure strain, PCR amplification is carried out by selecting bacterial 16SrDNA universal primers 27F and 1492R, and then sequencing analysis is carried out. The sequencing result is compared by BLAST in NCBI database, the strain is identified as Paracoccus denitrificans, named as Paracoccus densitificans TS-1, the strain is the deposited strain Paracoccus denitrificans Paracoccus densitificans TS-1, and is deposited in China general microbiological culture Collection center in 2019, 4 and 19 months, and the address is as follows: beijing, west way No.1 hospital on chaoyang district, no. 3, institute for microbiology, chinese academy of sciences, zip code: 100101, preservation number CGMCC No.17605.
The growth curve study of paracoccus denitrificans TS-1 is carried out, LB culture medium is selected for culture, and the result is shown in the attached figure 2: paracoccus denitrificans TS-1 can grow rapidly in an organic carbon source heterotrophic culture medium, and the bacterial liquid OD is obtained when the culture medium is cultured for 21 hours 600 The value reaches 1.0288, and the bacterial liquid OD is obtained after 42h 600 The value was maintained around 1.23 and the plateau was as long as 72h. The strain is propagated and increased quickly in a heterotrophic culture medium, and has a long stationary phase. In engineering application, an organic carbon source heterotrophic culture medium can be used for quickly obtaining high-density thalli, so that the method has strong advantages of efficiently and quickly removing ammonia nitrogen and sulfide in wastewater, can quickly form a membrane in a biological deodorization trickling filtration tower, is quickly started, and provides a better bacterial source for engineering deodorization.
Example 3: denitrification and desulfurization test Using the Denitrification and desulfurization Medium
100mL of denitrogenation and desulfurization medium solution is sterilized in a sterilization pot for 30min at the temperature of 115 ℃, then paracoccus denitrogenation bacterial liquid cultured for 24h is inoculated according to the inoculation amount of 5%, the bottle mouth is plugged by an aerobic plug, and the culture medium solution is placed in a shaking table to shake (30 ℃, 200 r/min).
Adding sterile water with the same amount as the inoculation amount into a control group, setting each experimental group for three times, and measuring the concentrations of ammonia nitrogen, nitrate nitrogen and nitrite nitrogen every 2 hours; sampling every 0.5h to determine the concentration of sulfide and elemental sulfur, and averaging the results.
Compared with a blank control group, the removal efficiency of ammonia nitrogen in the culture medium added with the TS-1 strain is obviously improved, the ammonia nitrogen degradation rate reaches 95.35% in 12 hours, the ammonia nitrogen concentration in the culture medium of the blank control group is not obviously changed, the removal rate is only 0.27%, and the strain has strong ammonia nitrogen degradation capability.
The initial nitrate nitrogen of the culture medium added with the TS-1 strain is about 70mg/L, the concentration of the nitrate nitrogen can be reduced to 4mg/L within 12 hours, and the degradation rate of the nitrate nitrogen is 94.28%. The nitrate nitrogen concentration in the blank control group does not decrease but slightly increases within 12h, which proves that the denitrification effect of the bacteria is remarkable.
The concentration of sulfide in the culture medium added with the TS-1 strain is obviously reduced compared with that of illumination, the sulfide removal rate reaches 53% in 6 hours, and the sulfide removal rate reaches 98% in 12 hours. The conversion rate of the elemental sulfur in the TS-1 culture medium reaches 60% after 4.5h, and the maximum elemental sulfur generation amount can reach 486.58mg/L. Comparing the conversion rate of sulfide and elemental sulfur in the blank control group, it can be known that the removal of sulfide in the blank control group is only simple physical removal and is not biotransformation removal, and the sulfide is subjected to valence state change in the culture medium inoculated with paracoccus denitrificans, which proves that the sulfide participates in biological reaction.
Example 4: performing denitrification and desulfurization test by using denitrification and desulfurization culture medium II
Sterilizing 100mL of the denitrogenation and desulfurization medium solution II in a sterilization pot at 115 ℃ for 30min, inoculating paracoccus denitrogenation bacterial liquid cultured for 24h according to the inoculation amount of 5%, plugging a bottle mouth with an aerobic plug, placing in a shaking table for shaking (30 ℃, 200 r/min), and determining that the initial ammonia nitrogen concentration is 170mg/L and the initial sulfide concentration is 556mg/L.
Adding sterile water with the same amount as the inoculation amount into a control group, setting each experimental group for three times, and measuring the concentrations of ammonia nitrogen, nitrate nitrogen and nitrite nitrogen every 2 hours; sampling every 0.5h to determine the concentration of sulfide and elemental sulfur, and averaging the results.
The results of the measurement after 12 hours are shown in FIGS. 3 to 6.
Compared with a blank control group, the removal efficiency of ammonia nitrogen in the culture medium added with the TS-1 strain is obviously improved, the degradation rate of the ammonia nitrogen reaches 99.6 percent in 12 hours, the concentration of the ammonia nitrogen in the culture medium of the blank control group is not obviously changed, the removal rate is only 0.27 percent, and the strain has stronger capability of degrading the ammonia nitrogen.
FIG. 4 shows the variation of various factors of paracoccus denitrificans TS-1 in the experiment process of degrading nitrate nitrogen. The initial nitrate nitrogen of the culture medium added with the TS-1 strain is about 170mg/L, the concentration of the nitrate nitrogen can be reduced to 3mg/L within 10 hours, and the degradation rate of the nitrate nitrogen is as high as 98.23%. The nitrate nitrogen concentration in the blank control group does not decrease within 10h but slightly increases, which proves that the bacteria have remarkable denitrification effect.
FIG. 5 shows the degradation capability of TS-1 on sulfides in the culture medium, and it can be seen that the sulfide concentration is significantly lower than that of illumination, the removal rate of sulfides reaches 58% in 6h, and the removal rate of sulfides reaches 100% in 12 h. In addition, in the attached figure 6, the conversion rate of the elemental sulfur in the TS-1 culture medium reaches 66.7% in 4.5h, and the maximum elemental sulfur generation amount can reach 523.46mg/L. Comparing the conversion rate of sulfide and elemental sulfur in the blank control group, it can be known that the removal of sulfide in the blank control group is only simple physical removal and is not biotransformation removal, and the sulfide is subjected to valence state change in the culture medium inoculated with paracoccus denitrificans, which proves that the sulfide participates in biological reaction.
By combining the shake flask experiment in example 3, it can be found that paracoccus denitrificans TS-1 has not only higher capabilities of heterotypic nitrification and aerobic denitrification, but also stronger capability of removing sulfides, and is a strain with three functions.
Example 5: deodorization experiment of biological trickling filter
Culturing paracoccus denitrificans TS-1 in an LB culture medium for 24 hours to obtain paracoccus denitrificans TS-1 bacterial liquid. In the process of culturing paracoccus denitrificans TS-1 in an LB culture medium, the culture temperature is 27 ℃, the pH value of the culture medium is 6.0, and the C/N in the culture medium is 12.
Adding a circulating culture medium into a biological trickling filter filled with polyhedral hollow ball filler, and circularly injecting 2L of paracoccus denitrificans TS-1 bacterial liquid cultured for 24 hours into the biological trickling filter filled with the filler by using a peristaltic pumpTower with 4L/min air flow rate and gas concentration of 40mg/m 3 The gas inflow and the gas concentration of the hydrogen sulfide are 40mg/m 3 The ammonia gas inflow of the biological trickling filter tower is simultaneously introduced with air, hydrogen sulfide and ammonia gas, the environmental temperature is kept at 25 ℃, the pH value is 6.5, and the humidity is 38 percent, and the bacteria can be stably attached to the filler by film hanging and domestication for 6 days under the condition. And then introducing ammonia gas and hydrogen sulfide gas into the biotrickling filter to perform a verification test for removing the ammonia gas and the hydrogen sulfide. And adding a circulating culture medium into the bio-trickling filter during a verification experiment process for removing ammonia and hydrogen sulfide by introducing ammonia and hydrogen sulfide gas, wherein the circulating culture medium provides nutrients required by growth for paracoccus denitrificans TS-1.
In the invention, the biofilm domestication is to generate a layer of microbial film which can adapt to and process hydrogen sulfide and ammonia gas on the surface of a polyhedral hollow filler.
Blank filler was added to the control group and no strain biofilm formation experiments were performed. Experiments were performed with tap water instead of the circulating liquid. And (3) introducing ammonia gas and hydrogen sulfide with the same concentration when the film hanging of the experimental group is finished. Then respectively recording the gas concentration at the inlet and the outlet of the two towers, thereby observing the removal and degradation capability of paracoccus denitrificans TS-1 on gas pollutants. The results are shown in the following table.
TABLE 3 deodorant Effect of Paracoccus denitrificans TS-1 in example 5
| Project index | Control group | Experimental group |
| Removal rate of Ammonia (%) | 27.22 | 100.0 |
| Hydrogen sulfide removal Rate (%) | 17.23 | 96.34 |
Example 6: biological trickling filter deodorization experiment two
Culturing paracoccus denitrificans TS-1 in an LB culture medium for 24 hours to obtain paracoccus denitrificans TS-1 bacterial liquid. In the process of culturing paracoccus denitrificans TS-1 in an LB culture medium, the culture temperature is 30 ℃, the pH value of the culture medium is 6.5, and the C/N in the culture medium is 15.
Adding a circulating culture medium into a biological trickling filter tower filled with polyhedral hollow sphere filler, and circularly injecting 2L of paracoccus denitrificans TS-1 bacterial liquid cultured for 24 hours into the biological trickling filter tower filled with the filler by using a peristaltic pump, wherein the air flow rate and the gas concentration are 5L/min and 50mg/m 3 The gas inflow and the gas concentration of the hydrogen sulfide are 50mg/m 3 The air, hydrogen sulfide and ammonia gas are simultaneously introduced into the biological trickling filter, the environmental temperature is kept at 26 ℃, the pH value is 7, and the humidity is 41 percent, and the bacteria can be stably attached to the filler by film hanging and domestication for 6 days under the condition. And then introducing ammonia gas and hydrogen sulfide gas into the biotrickling filter to perform a verification test for removing the ammonia gas and the hydrogen sulfide. And adding a circulating culture medium into the bio-trickling filter during a verification experiment process of introducing ammonia gas and hydrogen sulfide gas to remove the ammonia gas and the hydrogen sulfide, wherein the circulating culture medium provides nutrition required by growth for the paracoccus denitrificans TS-1.
In the invention, the film hanging domestication is to generate a layer of microbial film which can adapt to and process hydrogen sulfide and ammonia gas on the surface of a multi-surface hollow filler.
Blank filler was added to the control group and no strain biofilm formation experiments were performed. Experiments were performed with tap water instead of the circulating liquid. And (3) introducing ammonia gas and hydrogen sulfide with the same concentration when the film hanging of the experimental group is finished. Then respectively recording the gas concentration at the inlet and the outlet of the two towers, thereby observing the removal and degradation capability of paracoccus denitrificans TS-1 on gas pollutants. The results are shown in the following table.
TABLE 4 deodorizing Effect of Paracoccus denitrificans TS-1 in example 6
| Project index | Control group | Experimental group |
| Removal rate of Ammonia (%) | 27.22 | 100.0 |
| Hydrogen sulfide removal rate (%) | 17.23 | 97.36 |
Example 7: deodorization experiment III of biological trickling filter
Culturing paracoccus denitrificans TS-1 in an LB culture medium for 24 hours to obtain paracoccus denitrificans TS-1 bacterial liquid. During the culture of the paracoccus denitrificans TS-1 in the LB culture medium, the culture temperature is 33 ℃, the pH value of the culture medium is 7, and the C/N in the culture medium is 18.
Adding a circulating culture medium into a biological trickling filter tower filled with polyhedral hollow sphere filler, and circularly injecting 2L of paracoccus denitrificans TS-1 bacterial liquid cultured for 24 hours into the biological trickling filter tower filled with the filler by using a peristaltic pump, wherein the air flow rate and the gas concentration are 6L/min and 60mg/m 3 The gas inflow and the gas concentration of the hydrogen sulfide are 60mg/m 3 The air, hydrogen sulfide and ammonia gas are simultaneously introduced into the biological trickling filter, the environmental temperature is kept at 27 ℃, the pH value is 7.5, and the humidity is 45 percent, and the bacteria can be stably attached to the filler by film hanging and domestication for 7 days under the condition. Then, the ammonia gas and the hydrogen sulfide gas are introduced into the biotrickling filter to remove the ammonia gas and the hydrogen sulfideAnd (4) testing. And adding a circulating culture medium into the bio-trickling filter during a verification experiment process of introducing ammonia gas and hydrogen sulfide gas to remove the ammonia gas and the hydrogen sulfide, wherein the circulating culture medium provides nutrition required by growth for the paracoccus denitrificans TS-1.
In the invention, the biofilm domestication is to generate a layer of microbial film which can adapt to and process hydrogen sulfide and ammonia gas on the surface of a polyhedral hollow filler.
Blank filler was added to the control group and no strain biofilm formation experiments were performed. Experiments were performed with tap water instead of the circulating liquid. And (3) introducing ammonia gas and hydrogen sulfide with the same concentration when the film hanging of the experimental group is finished. Then the gas concentrations at the inlet and the outlet of the two towers are respectively recorded, so that the removal and degradation capacity of the paracoccus denitrificans TS-1 on the gas pollutants is observed. The results are shown in the following table.
TABLE 5 deodorant Effect of Paracoccus denitrificans TS-1 in example 7
| Project index | Control group | Experimental group |
| Removal rate of Ammonia (%) | 27.22 | 100.0 |
| Hydrogen sulfide removal Rate (%) | 17.23 | 95.93 |
By combining the deodorization tests of the embodiments 5 to 7, it can be found that the removal rate of paracoccus denitrificans TS-1 to ammonia in odor can reach 100%, the removal rate to hydrogen sulfide gas can reach 97.36% at most, the deodorization effect is good, and toxic and harmful gases such as hydrogen sulfide and ammonia can be effectively removed, so that the deodorant is applied to deodorization and sewage treatment.
And that the present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Sequence listing
<110> university of west safety engineering
Chengdu Institute of Biology, Chinese Academy of Sciences
<120> Paracoccus denitrificans strain screening and application thereof in deodorization
<160> 1
<170> SIPOSequenceListing 1.0
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<211> 1327
<212> DNA
<213> Paracoccus denitrificans (Paracoccus denitirichicans TS-1)
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tcgctgcctc cattgctggt tagcgcacgg ccgtcgggta gacccaactc ccatggtgtg 60
acgggcggtg tgtacaaggc ccgggaacgt attcaccgcg gcatgctgtt ccgcgattac 120
tagcgattcc aacttcatgg ggtcgagttg cagaccccaa tccgaactga gatggctttt 180
ggggattaac ccactgtcac caccattgta gcacgtgtgt agcccaaccc gtaagggcca 240
tgaggacttg acgtcatcca caccttcctc cgacttatca tcggcagttc tcttagagtg 300
cccaaccaaa tgctggcaac taagagtgtg ggttgcgctc gttgccggac ttaaccgaac 360
atctcacgac acgagctgac gacagccatg cagcacctgt ccacaggtct cttacgagaa 420
aactccatct ctggagcggt cctgcgatgt caagggttgg taaggttctg cgcgttgctt 480
cgaattaaac cacatgctcc accgcttgtg cgggcccccg tcaattcctt tgagttttaa 540
tcttgcgacc gtactcccca ggcggaatgc ttaatccgtt aggtgtgtca ccgaacagca 600
tgctgcccga cgactggcat tcatcgttta cggcgtggac taccagggta tctaatcctg 660
tttgctcccc acgctttcgc acctcagcgt cagtatcgag ccagtgagcc gccttcgcca 720
ctggtgttcc tccgaatatc tacgaatttc acctctacac tcggaattcc actcacctct 780
ctcgaactcc agaccgatag ttttgaaggc agttccgagg ttgagccccg ggatttcacc 840
cccaactttc cggtccgcct acgtgcgctt tacgcccagt aattccgaac aacgctagcc 900
ccctccgtat taccgcggct gctggcacgg agttagccgg ggcttcttct gctggtaccg 960
tcattatctt cccagctgaa agagctttac aaccctaggg ccttcatcac tcacgcggca 1020
tggctagatc agggttgccc ccattgtcta agattcccca ctgctgcctc ccgtaggagt 1080
ctgggccgtg tctcagtccc agtgtggctg atcatcctct caaaccagct atggatcgtc 1140
ggcttggtag gccattaccc caccaactac ctaatccaac gcgggctaat cctttgccga 1200
taaatctttc ccccaaaggg cgtatacggt attactccca gtttcccggg gctattccgt 1260
agcaaagggc atattcccac gcgttactca cccgtccgcc gctaaccccg aagggtcgct 1320
cgactgc 1327
Claims (7)
1. Paracoccus denitrificansParacoccus denitrificans) The method is characterized in that: the strain name is as follows: paracoccus denitrificans (A)Paracoccus denitrificans) TS-1, deposited in China general microbiological culture Collection center (CGMCC) at 19.4.2019 with the deposit number: CGMCC NO.17605.
2. The use of paracoccus denitrificans according to claim 1 for deodorization.
3. The use of paracoccus denitrificans in deodorization according to claim 2, comprising the following specific steps:
(1) Culturing paracoccus denitrificans TS-1 into paracoccus denitrificans TS-1 bacterial liquid, wherein the active bacterial quantity of the paracoccus denitrificans TS-1 in the bacterial liquid is 10 8 ～10 9 CFU/mL；
(2) Adding a solution which provides nutrition required by growth of paracoccus denitrificans TS-1 into a biological trickling filter filled with a filler, and then adding the bacterial liquid of the paracoccus denitrificans TS-1 obtained in the step (1);
(3) The air flow rate of 4 to 6L/min and the gas concentration of 40 to 60mg/m 3 The gas inflow and the gas concentration of the hydrogen sulfide are 40-60 mg/m 3 Introducing air, hydrogen sulfide and ammonia gas into the biological trickling filter simultaneously, and performing biofilm culturing and domestication for 6-8 days under the conditions of the ambient temperature of 25-27 ℃, the pH value of 6.5-7.5 and the humidity of 38% -45%;
(4) And after the membrane hanging acclimation is finished, introducing gas to be treated into the biological trickling filtration tower.
4. The use of paracoccus denitrificans according to claim 3 for deodorization, wherein: in the step (1), the paracoccus denitrificans TS-1 is cultured into the paracoccus denitrificans TS-1 bacterial liquid under the conditions that the culture temperature is 27-33 ℃, the pH value of the culture medium is 6.0-7.0, and the C/N ratio in the culture medium is 12-18.
5. The use of paracoccus denitrificans according to claim 4 for deodorization, wherein: in the step (1), the paracoccus denitrificans TS-1 is cultured into the paracoccus denitrificans TS-1 bacterial liquid under the conditions that the culture temperature is 30 ℃, the pH value of the culture medium is 6.5 and the C/N ratio in the culture medium is 15.
6. Use of paracoccus denitrificans according to claim 3 for deodorization, wherein: and (4) adding a solution which provides required nutrition for growth of paracoccus denitrificans TS-1 into the biological trickling filter while introducing the gas to be treated.
7. The use of paracoccus denitrificans according to claim 1 in sewage treatment.
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| CN110452846B (en) * | 2019-08-16 | 2022-07-12 | 鹭滨环保科技（上海）股份有限公司 | Paracoccus denitrificans and biological denitrification method by combining paracoccus denitrificans with mineralization bed |
| CN114181856B (en) * | 2021-11-30 | 2023-12-15 | 南京农业大学 | Microbial combination capable of effectively inhibiting odor production of biomass fermentation from source and application thereof |
| CN114149944B (en) * | 2021-11-30 | 2024-04-09 | 南京农业大学 | Microorganism combination capable of efficiently adsorbing malodorous gas at tail end, enhanced colonization and application |
| CN114958689B (en) * | 2022-06-29 | 2023-03-10 | 山东碧蓝生物科技有限公司 | Paracoccus, microbial inoculum and application thereof in livestock and poultry breeding deodorization field |
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