Salt-tolerant amide organic matter degradation bacterial agent and its application

Abstract

The present invention provides a salt-tolerant amide organic matter degrading bacterium and its application, wherein the salt-tolerant amide organic matter degrading bacterium comprises: Paenarthrobacter sp.) P24; and / or Nitratireductor indicus N30; and / or Rhodococcus ( Rhodococcus sp.) R25, all of which are deposited in the General Microbiological Center of China Microbiological Culture Collection Committee, with the deposit numbers CGMCC No.25361, CGMCC No.25360 and CGMCC No.25362 respectively. The degradation bacterial agent of this technical solution can effectively treat wastewater containing amide organic matter in a high-salinity environment.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a salt-resistant amide organic matter degradation bacterial agent and application thereof. Background Art

[0002] Typical industries in my country, such as petrochemicals and pharmaceuticals, discharge a large amount of high-salt and high-concentration organic wastewater every year. Such wastewater contains not only high-concentration inorganic salts, but also a large number of refractory toxic organic pollutants, which seriously inhibit the activity of microorganisms in activated sludge and affect the efficiency and functional stability of traditional biological treatment. High-salt wastewater is usually defined as a salt content of at least 1wt%, and 5% of the total global wastewater is high-salt organic wastewater. The discharge of high-salt wastewater in my country is still increasing at a rate of 2% per year, which poses a huge challenge to sewage treatment plants. Due to the high concentration of inorganic salts and other toxic organic compounds (COD >2000 mg / L), direct discharge and inefficient treatment by sewage treatment plants may lead to the death of aquatic species and damage to the ecosystem, which seriously restricts the survival and sustainable development of enterprises. Therefore, it is urgent to develop green treatment methods for treating high-salt and high-concentration refractory organic wastewater.

[0003] Research on high-salinity organic wastewater mainly focuses on physicochemical methods (membrane separation, evaporation crystallization, advanced oxidation, etc.) and biological treatment. Among them, physicochemical treatment has disadvantages such as high operating costs, large energy consumption, and possible secondary pollution. In contrast, the use of microbial degradation to treat high-salinity organic wastewater is an economical, green and sustainable approach, which is of great significance to reducing the pollution of high-salinity organic wastewater to the environment and reducing the ecological risks caused by it. However, high salt concentrations usually cause an increase in the osmotic pressure of the water environment, leading to the separation of the cell wall of freshwater microorganisms, and also cause inorganic salt poisoning of biological enzymes, which reduces the ability of microorganisms to degrade pollutants.

[0004] As a common amide chemical intermediate, acetoacetanilide (AAA) can be used as a raw material for sulfonamide drugs and used to produce analgesics, antipyretics and preservatives. It is also an intermediate for the production of dyes, used to make pyrazolone, p-nitroacetanilide, p-nitroaniline and p-phenylenediamine. In agriculture, it can be used as a raw material for acetanilide herbicides such as carboxin. Pyrazolone (PMP) is mainly used as a raw material for the production of pharmaceuticals such as antipyrine, aminopyrine and analgin, and is also used in dyes and color film dyes, pesticides and organic synthesis industries. AAA and PMP, two typical amide compounds, have a wide range of uses and a large amount of use, and enterprises are facing the dilemma of the treatment of such amide organic wastewater that needs to be solved urgently. Summary of the invention

[0005] In response to the above technical problems, the present invention discloses a salt-tolerant amide organic matter degrading bacterial agent and its application, which can efficiently treat organic wastewater containing PMP and AAA in a high-salt environment, and has important practical significance for achieving efficient biological treatment of high-salt difficult-to-degrade organic wastewater.

[0006] To this end, the technical solution adopted by the present invention is:

[0007] Salt-tolerant amide organic matter degrading bacteria, including:

[0008] Arthrobacter Paenarthrobacter sp.) P24, deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, deposit date: July 20, 2022; deposit number: CGMCC No. 25361;

[0009] and / or Nitratireductor indicus N30, deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, deposit date: July 20, 2022; deposit number: CGMCC No. 25360;

[0010] and / or Rhodococcus ( Rhodococcus sp.) R25, deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit date is July 20, 2022; the deposit number is CGMCC NO. 25362.

[0011] The main biological characteristics of the three pure strains are: Paenarthrobacter sp. P24 is a Gram-positive bacterium, classified as Paenarthrobacter sp. , the colonies on R2A medium are light yellow, opaque, round colonies with regular edges. Rhodococcus sp. R25 is a Gram-positive bacterium, classified and named Rhodococcus erythropolis On R2A medium, the colonies are goose yellow, opaque, and circular with regular edges. Nitratireductor indicus N30 is a Gram-negative bacterium, classified as Nitratireductor indicus , in an area suitable for its growth Nitratireductor The colonies on the culture medium are white, opaque, round colonies with regular edges.

[0012] As a further improvement of the present invention, the Rhodococcus Rhodococcus sp. R25, Arthrobacter Paenarthrobacter sp. P24 and Nitratireductor indicus The 16S rRNA gene sequences of N30 are shown in SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3, respectively.

[0013] In the above technical solution, degradation experiments were carried out in an aerobic inorganic salt medium with an optimum temperature of 30°C and an optimum initial pH of 6.8-7.0. Rhodococcus sp. R25 completely degraded 100 mg / L AAA within 6 days, but could degrade nearly 90% of AAA within 2 days, and at the same time completely degraded aniline, the hydrolysis degradation product of AAA. Nitratireductor indicus N30 and Arthrobacter Paenarthrobacter sp. P24 has a removal rate of nearly 90% for AAA (100 mg / L) within 1 day. At the same time, Rhodococcus R25 is also an efficient ammonia-degrading bacterium for PMP, which can completely degrade 150 mg / L PMP within 3 days under 3% salinity conditions. Using one, two or three of the above three bacteria can effectively treat organic wastewater containing AAA in a high-salinity environment; using Rhodococcus R25 alone can effectively treat organic wastewater containing PMP.

[0014] The formula of the inorganic salt culture medium is preferably as follows: the composition of the only carbon and nitrogen source inorganic salt culture medium (per liter of water): NaCl 5g, MgSO4·7H2O0.2g, KH2PO4 0.5g, K2HPO4∙3H2O 1.5g, and the pH value is adjusted to 6.8-7.0.

[0015] R2A medium composition (per liter of water): yeast extract 0.5 g, tryptone 0.5 g, casein hydrolyzate 0.5 g, glucose 0.5 g, soluble starch 0.5 g, K2HPO4∙3H2O 0.3 g, MgSO4·7H2O 0.05 g, sodium pyruvate 0.3 g. Sterilize at 115℃ for 30 min.

[0016] As a further improvement of the present invention, the composition and volume ratio of the salt-tolerant amide organic matter degrading bacteria are: Paenarthrobacter sp.)P24 20-60%, Nitratireductor indicus N30 20-40%, Rhodococcus ( Rhodococcus sp.) R25 20-40%.

[0017] Using this technical solution, the synthetic bacterial community constructed by three pure bacteria was used to treat actual AAA wastewater when the dosage of the synthetic bacterial community / organic wastewater was 0.1wt‰. The AAA mother liquor was diluted 1 and 3 times (2000-3200 mg / L). After 14 days, the AAA removal rates reached 69.28±0.42% and 83.50±0.39%, and the TOC removal rates reached 55.34±0.12% and 65.90±0.33%, respectively. This shows that the synthetic bacterial community can treat the actual high-salt and high-concentration AAA mother liquor, providing degradation bacterial resources for the treatment of actual AAA mother liquor in industrial parks, and has good engineering application prospects.

[0018] As a further improvement of the present invention, the Arthrobacteroids ( Paenarthrobacter sp.)P24, Nitratireductor indicus N30, Rhodococcus Rhodococcus sp.) R25 was separated from activated sludge in a sewage treatment plant.

[0019] Nitratireductor indicus N30 separation medium composition (per liter of water): yeast extract 1.0 g, peptone 5.0 g, ferric citrate 0.1 g, MgCl2 8.8 g, Na2SO4 3.24 g, CaCl2 1.8 g, KCl 0.55 g, NaHCO3 0.16 g, KBr 0.08 g, SrCl2 0.034 g, Na2SiO3 0.004 g, boric acid 0.022 g, NaF 0.0024 g, NH4NO3 0.0016 g, Na2HPO4∙3H2O 0.008 g. Sterilize at 121°C for 30 min.

[0020] As a further improvement of the present invention, the Arthrobacteroids ( Paenarthrobacter sp.)P24, Nitratireductor indicus N30, Rhodococcus Rhodococcus sp.) R25 can use AAA as the only carbon and nitrogen source and energy under aerobic conditions, assimilate AAA for its own growth and reproduction, accumulate the degradation product aniline, and continue to use the degradation product AN as a carbon and nitrogen source and energy for its own growth and reproduction, thereby degrading AAA.

[0021] As a further improvement of the present invention, the Rhodococcus ( Rhodococcus sp.) R25 uses PMP as the only nitrogen source and achieves ammoniation of high-salinity and high-concentration PMP wastewater through aerobic or anaerobic assimilation of PMP.

[0022] As a further improvement of the present invention, the amide organic matter includes at least one of AAA and PMP.

[0023] The present invention also discloses the application of the salt-tolerant amide organic matter degrading bacteria as described above, which is used for degrading AAA / PMP in amide-containing organic wastewater.

[0024] As a further improvement of the present invention, the salt-tolerant amide organic matter-degrading bacteria are applied to the degradation of residual amide organic matter AAA / PMP in sewage, substrate and soil.

[0025] As a further improvement of the present invention, the degradation environment of the salt-tolerant amide organic matter degrading bacteria is: in an environment with a salt content of 0.1-5%, a temperature of 15-40° C., an initial pH of 5-9, and under aerobic conditions.

[0026] As a further improvement of the present invention, the dosage of the salt-tolerant amide organic matter degrading bacteria is 0.1 to 1 wt‰ of the amount of organic wastewater.

[0027] As a further improvement of the present invention, the seed liquid of the salt-tolerant amide organic matter degrading bacteria is cultured separately to obtain a concentrated bacterial liquid which is directly added for use, or three strains of synthetic bacteria are cultured together to obtain a concentrated bacterial liquid or made into a dry powdered bacterial agent using existing technology in the field for use.

[0028] The present invention also discloses a method for treating wastewater containing high-salt and high-concentration amide organic matter, which is treated by using the above-mentioned salt-tolerant amide organic matter degrading bacterial agent. Further preferably, the amide organic matter includes AAA and / or PMP.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] First, the technical solution of the present invention can effectively treat organic wastewater containing PMP and AAA in a high-salt environment. Rhodococcus sp. R25 could completely degrade 100 mg / L AAA within 6 days, and degrade nearly 90% of AAA within 2 days, while also achieving complete degradation of aniline, the hydrolysis product of AAA. Nitratireductor indicus N30 and Arthrobacter Paenarthrobacter sp. P24 had a degradation rate of nearly 90% on the first day of AAA within 1 day. The synthetic bacterial community constructed by three pure bacteria had a removal rate of more than 90% for AAA (100 mg / L) within 1 day, and the degradation was complete in 3 days. Moreover, the degradation rate of the synthetic bacterial community was faster than that of each pure bacteria. In addition, Rhodococcus R25 could efficiently degrade PMP under salinity conditions of up to 3%, realize ammoniation of it, and expand its substrate spectrum for degrading amide organic matter.

[0031] Second, the three strains of high-salt and high-concentration AAA / PMP-degrading bacteria disclosed in the technical solution of the present invention can efficiently treat high-salt and high-concentration AAA wastewater after constructing a synthetic bacterial community, and can tolerate a salinity range of up to 5%. The high-quality pure bacteria Rhodococcus R25 can also tolerate a salinity range of up to 5% in treating PMP wastewater.

[0032] Third, the three strains of high-salt and high-concentration AAA / PMP degradation bacteria disclosed in the technical solution of the present invention are all isolated from the activated sludge of the sewage treatment plant, which can better adapt to engineering applications under natural conditions. Compared with physical and chemical methods, the cost is greatly reduced, and it is green and has no toxic by-products. It has important engineering significance for the biological enhancement treatment of high-salt and high-concentration AAA / PMP actual wastewater. So far, there has been no report on the use of pure bacteria and synthetic bacterial flora agents to treat high-salt and high-concentration AAA / PMP wastewater. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 The following are photos of pure colonies of three strains of high-salt and high-concentration AAA or PMP-degrading bacteria of the present invention; (a) is a photo of the colonies of Rhodococcus R25 on an R2A plate, (b) is a photo of the colonies of Arthrobacter P24 on an R2A plate, and (c) is a photo of the colonies of Nitratireductor indicus N30 is suitable for Nitratireductor Photograph of colonies growing on a growth medium plate.

[0034] Figure 2 The figures are scanning electron microscope and transmission electron microscope photographs of the high-salt and high-concentration AAA or PMP degrading bacteria of the present invention, wherein (a) and (b) are scanning electron microscope and transmission electron microscope photographs of strain R25, respectively; (c) and (d) are scanning electron microscope and transmission electron microscope photographs of strain P24, respectively; (e) and (f) are scanning electron microscope and transmission electron microscope photographs of strain N30, respectively.

[0035] Figure 3 The strain constructed based on 16S rRNA gene of the present invention Rhodococcus. sp.R25, Paenarthrobacter sp. P24 and Nitratireductor indicus Phylogenetic tree of N30.

[0036] Figure 4 The following are the validation diagrams of the degradation efficiency of the three pure bacteria of the present invention on AAA (100 mg / L), wherein (a) is Nitratireductor indicus N30, (b) Paenarthrobacter sp. P24, (c) Rhodococcus sp. R25.

[0037] Figure 5 The salt tolerance verification diagram of the three pure bacteria of the present invention, wherein (a) is Nitratireductor indicus N30, (b)Paenarthrobacter sp. P24, (c) Rhodococcus sp. R25.

[0038] Figure 6 The effect of the synthetic bacterial community of the present invention at 1% salinity, wherein (a) is the change curve of AAA, AN and biomass, (b) is the change curve of TN and NH3-N, and (c) is the change curve of TOC.

[0039] Figure 7 These are the effects of the synthetic bacterial flora of the present invention on AAA mother liquor diluted 1 and 3 times, where (a) shows AAA degradation and AN generation, (b) shows COD and TOC, and (c) shows the changes in TN and NH3-N.

[0040] Figure 8 The sedimentation of the Rhodococcus sp. R25 cultured to the late logarithmic phase after being left to stand for 0 h (a), 3 h (b) and 6 h (c) and the transmission electron microscopic characteristics of the bacterial cells wrapped by the extracellular polymer EPS (d)

[0041] Figure 9 The curves are the degradation and amination of the amide heterocyclic compound PMP by the Rhodococcus sp. R25 of the present invention under high salt conditions. DETAILED DESCRIPTION

[0042] The present invention is described below in conjunction with the accompanying drawings and specific embodiments. Adjustments to some methods and parameters in the present invention without departing from the spirit and essence of the present invention are deemed to fall within the scope of the present invention.

[0043] The chemical reagents used in the implementation of this example are all commercially available analytically pure or above. Standards such as AAA and PMP and reagents used in the inorganic salt culture medium are purchased from Aladdin Reagent Company (Shanghai). The components of the inorganic salt culture medium are as described in the content of the invention and are sterilized by high temperature and high pressure.

[0044] Example 1

[0045] Isolation and identification of AAA-degrading bacteria.

[0046] 1.1 Isolation and purification of pure degradation bacteria

[0047] The sludge from the secondary sedimentation tank of Harbin Wenchang Wastewater Treatment Plant was used as the inoculum source for the isolation of salt-tolerant degrading bacteria, and the target AAA was used as the only carbon and nitrogen source and energy source for enrichment and domestication. The enrichment medium used was 100 mL of inorganic salt medium (containing 0.5% salt and an initial AAA concentration of 150 mg / L) sterilized at high temperature. About 5 g of activated sludge was inoculated in a 250 mL conical flask and cultured in a shaker at 30°C and 150 rpm. Every 7 days, the inoculum was transferred to the same fresh enrichment medium at a 5% inoculum. After 5 consecutive transfers, the enrichment solution in the conical flask was able to grow with AAA as the only carbon and nitrogen source and energy source, and presented a uniform white suspension (without granular sludge or other impurities). At this time, the AAA degradation efficiency of the enrichment solution can be preliminarily evaluated by ultra-high performance liquid chromatography (UPLC), total organic carbon (TOC) and biomass changes (growth curve). After enrichment and acclimatization, we finally obtained an enrichment solution that can grow and degrade AAA as the only carbon and nitrogen source. When the initial concentration of AAA was 150 mg / L, the enrichment solution could be completely degraded in two days. Under the condition of salinity of 5%, the enrichment solution could still efficiently and stably degrade AAA. This proves that the obtained enrichment solution has good salt tolerance potential. Nitratireductor The strains were isolated and purified by dilution coating method and streak separation method. The results showed that the strains isolated from R2A medium Rhodococcus sp . and Paenarthrobacter sp., named R25 and P24 respectively. Nitratireductor Isolated from the growth medium Nitratireductor indicus The streaking operation was repeated several times, and the three purified strains were placed in an inorganic salt medium to determine their ability to degrade AAA. After the function was verified, they were stored in a -80°C refrigerator with glycerol.

[0048] Among them, the formula of inorganic salt culture medium is: the composition of the only carbon and nitrogen source inorganic salt culture medium (per liter of water): NaCl 5g, MgSO4·7H2O0.2 g, KH2PO4 0.5 g, K2HPO4∙3H2O 1.5 g, and the pH value is adjusted to 6.8-7.0.

[0049] R2A medium composition (per liter of water): 0.5 g yeast extract, 0.5 g tryptone, 0.5 g casein hydrolyzate, 0.5 g glucose, 0.5 g soluble starch, 0.3 g K2HPO4∙3H2O, 0.05 g MgSO4·7H2O, 0.3 g sodium pyruvate. Sterilize at 115°C for 30 min.

[0050] Suitable Nitratireductor indicusThe composition of the medium for N30 growth (per liter of water): yeast extract 1.0 g, peptone 5.0 g, ferric citrate 0.1 g, MgCl2 8.8 g, Na2SO4 3.24 g, CaCl2 1.8 g, KCl0.55 g, NaHCO30.16 g, KBr 0.08 g, SrCl2 0.034 g, Na2SiO3 0.004 g, boric acid 0.022 g, NaF0.0024 g, NH4NO3 0.0016 g, Na2HPO4∙3H2O 0.008 g. Sterilize by high temperature and high pressure at 121°C for 30 min.

[0051] 1.2 Identification of the systematic taxonomic status of core strains R25, P24, and N30

[0052] Three strains of bacteria that can degrade AAA were isolated from the enrichment solution, and the isolated and purified strains were subjected to simple physiological and biochemical analysis and 16S rRNA gene sequencing. Figure 1 and Figure 2 As shown in Figure 1, the main biological characteristics of strain R25 are: Gram staining positive, short rod-shaped bacteria, goose yellow colonies on R2A medium, opaque, round colonies with regular edges. Its sequence is highly similar to Rhodococcu The 16S rRNA gene sequence is shown in the sequence table SEQ.NO.1, and the phylogenetic tree is shown in the attached Figure 3 Combined with its physiological and biochemical characteristics, the bacterium was identified as Rhodococcu sp., named Rhodococcu sp.R25. The main biological characteristics of strain P24 are: Gram staining positive, short rod-shaped bacteria, light yellow, opaque, round colonies with regular edges on R2A medium. Its sequence is highly similar to Paenarthrobacter The 16S rRNA gene sequence is shown in the sequence table SEQ.NO.2, and the phylogenetic tree is shown in the attached Figure 3 Combined with its physiological and biochemical characteristics, the bacterium was identified as Paenarthrobacter sp., named Paenarthrobacter sp.P24. The main biological characteristics of strain N30 are: Gram staining reaction is negative, the bacteria are short rod-shaped, and they can grow in a suitable environment. Nitratireductor The colonies on the culture medium are white, opaque, and circular with regular edges. Its sequence is highly similar to Nitratireductor The 16S rRNA gene sequence is shown in the sequence table SEQ.NO.3, and the phylogenetic tree is shown in the attached Figure 3 Combined with its physiological and biochemical characteristics, the bacterium was identified as Nitratireductor indicus , named Nitratireductor indicusN30.

[0053] Example 2

[0054] The degradation efficiency of the three pure bacteria isolated in Example 1 on AAA was tested.

[0055] 2.1 Preparation of pure bacterial suspension

[0056] The three purified strains isolated in Example 1 were respectively cultured in R2A solid medium supplemented with an additional carbon source (AAA) and a suitable Nitratireductor The culture medium (the culture medium formula is as described above) was activated, and then the pure colonies were picked to sterile LB medium and cultured on a shaker at 30°C and 150 rpm. After the bacterial solution became turbid, the bacterial solution in the logarithmic phase was centrifuged and washed with inorganic salt medium to prepare the bacterial suspension of each pure bacteria for subsequent verification of the degradation efficiency of the pure bacteria.

[0057] 2.2 Degradation efficiency verification experiment of three pure bacteria strains.

[0058] In order to verify the degradation efficiency of the obtained pure bacteria on AAA, 50 mg / L of substrate AAA was added to a 250 mL conical flask (containing 100 mL of sterile inorganic salt medium), and the prepared pure bacterial suspension was added to ensure that the initial OD value of each pure bacteria inoculated was 0.05. The culture was cultured at a constant temperature of 30°C with shaking. Each treatment was repeated 3 times, and the culture medium without bacterial solution was used as the control. The degradation efficiency of each pure bacteria on 100 mg / L AAA is shown in the figure. Figure 4 shown. Nitratireductor indicus N30 and Paenarthrobacter sp. P24 did not completely degrade AAA. However, the degradation rate of these two strains was as high as 90% within 1 day. Figure 4 (a) and 4(b). Rhodococcus sp. R25 required 6 days to completely degrade 100 mg / L AAA, but it could degrade nearly 90% of AAA within 2 days, and the hydrolysis product aniline generated during the degradation of AAA was also completely degraded. Figure 4 (c) shown.

[0059] Example 3

[0060] Experiment on degradation efficiency of pure bacteria under different salinities.

[0061] In order to verify the degradation effect of the obtained strains on AAA under different salinities, under the premise that the initial inoculation OD was 0.05, the strains N30, P24 and R25 were respectively subjected to degradation verification under the conditions of salt content of 0.1%, 1%, 3% and 5% and initial AAA concentration of 100 mg / L. The results are shown in Figure 2. Figure 5The experiment found that strain N30 did not completely degrade AAA, even at low salinity. The degradation rate of AAA within 7 days at 0.1% salinity was close to 90%. As the salinity increased, its degradation ability was affected by salinity. At 1% salinity, although the activity of strain N30 was inhibited, its degradation rate of AAA within 7 days was almost the same as that of 0.1% salinity. At 3% and 5% salinity, its degradation rate of AAA at 100 mg / L was 61.82% and 55.97%, respectively. Figure 5 (a) As shown. The degradation ability of strain N30 under high salt conditions provides strain resources for the later combination of functional bacteria. In the salinity range of 0.1-3%, the degradation ability of strain P24 on AAA was not affected by the increase of salinity. The degradation rate of 100 mg / L AAA within 7 days was close to 95%, and the removal rate was close to 95% on the first day. As the salinity increased to 5%, the degradation activity of strain P24 was inhibited to a certain extent in the first two days, but the degradation rate of 100 mg / L AAA within 7 days was still as high as 93%, as shown in Figure 2. Figure 5 (b) As shown. The high efficiency of AAA degradation demonstrated by strain 24 provides us with high-quality strain resources. Under 0.1% salinity conditions, AAA (100 mg / L) was completely degraded by strain R25 within 7 days. Within 7 days, the removal rates of AAA at 1% and 3% salinity reached 93.02±0.97% and 93.06±0.38%, respectively. Although the activity of the bacteria was partially inhibited at 5% salinity, the removal rate of AAA still reached 80.05±5.53%. Figure 5 (c) shown.

[0062] Example 4

[0063] Using one, two or three of the above three bacteria can effectively treat organic wastewater containing AAA in a high-salinity environment. The following mainly describes the synthetic bacterial flora containing the above three bacteria.

[0064] Experiment to verify the effectiveness of synthetic bacterial flora in treating simulated AAA wastewater.

[0065] Arthrobacter Paenarthrobacter sp. P24, Nitratireductor indicus N30 and Rhodococcus Rhodococcus sp. R25, the volume ratio of each strain is: strain P24 is 50%, strain N30 is 25%, strain R25 is 25%. The sum of the addition ratios of each strain is 100%. The synthetic bacterial flora was added into AAA wastewater.

[0066] Under 1% salinity conditions, the synthetic bacterial community (initial inoculation OD was 0.05) degraded more than 90% within 1 day and completely degraded within 3 days. Figure 6 (a) As shown. The maximum accumulation of AN degradation product of synthetic bacteria was 49.79 ± 1.15 mg / L, and the AN generated in 4 days could be completely degraded. After treatment with synthetic bacteria, TN decreased from 10.96 ± 0.23 mg / L to 4.75 ± 0.45 mg / L, and the removal rate reached 56.68 ± 0.08%, and the maximum accumulation of NH3–N was 4.38 ± 0.18 mg / L at 7 days. Figure 6 (b). And the TOC of the synthetic bacteria decreased from the initial 88.36±0.62mg / L to 10.2±0.51mg / L within 7 days, and the removal rate reached 88.45±0.28%. Figure 6 (c) shown.

[0067] Example 5

[0068] Based on Example 4, this example conducts an experiment to verify the effectiveness of treating actual AAA mother liquor with a synthetic bacterial flora.

[0069] Using the 1% salinity compounding scheme, the actual wastewater treatment experiment was carried out with an initial inoculation OD of 0.1 (synthetic bacterial community dosage / organic wastewater volume of 0.1wt‰). The volume ratio of each strain was: strain P24 was 60%, strain N30 was 20%, and strain R25 was 20%. The sum of the addition ratios of each strain was 100%, and it was tested whether the synthetic bacterial community solution could still play a degradation role in the high-salt and high-concentration environment of the actual AAA mother liquor. The degradation was verified by adding the synthetic bacterial community solution to the actual AAA mother liquor (diluted 1 times and 3 times, respectively). After the AAA mother liquor was diluted 1 times and 3 times, the AAA removal rates were 69.28% and 77.74% after 14 days, respectively. Figure 7 (a) shows that the TOC removal rates are 55.34% and 65.90%, and the COD removal rates are 40.51% and 61.18%, respectively. Figure 7 As shown in (b). After AAA mother liquor was diluted 1 and 3 times for 14 days, TN concentration decreased from the initial concentration of 399±1.41 mg / L and 252±1.41 mg / L to 328±2.83 mg / L and 199.5±10.61 mg / L, respectively. The TN removal rates were 17.79% and 20.83%, respectively. NH3-N increased from the initial 21.00±1.41 mg / L and 24.00±2.83 mg / L to the maximum of 84.00±2.83 mg / L and 64.00±2.83 mg / L, respectively. Figure 7 (c) shown.

[0070] It can be seen from the above experimental data that the verification experiment of using synthetic bacterial flora for AAA mother liquor treatment shows that the proposed compounding scheme is feasible in actual mother liquor treatment.

[0071] Example 6

[0072] Rhodococcus Rhodococcus sp. R25 degradation and amination of amide compounds PMP efficiency experiment.

[0073] The inoculation source is the pure bacterial suspension prepared in 2.1. The experiment found that Rhodococcus R25 was cultured in LB liquid medium to the middle and late logarithmic phase, and then left to settle for 6 hours to form a flocculent bacterial precipitate, as shown in Figures 8 (a), 8 (b), and 8 (c). The bacterial precipitate was examined by transmission electron microscopy (TEM), as shown in Figure 8 (d). It was found that the reason for the formation of flocculent precipitation was that the strain itself secreted extracellular polymers (EPS), and the EPS was connected and overlapped to form flocculent bacteria, thereby promoting the formation of bacterial flocs. Therefore, Rhodococcus R25, as a strain that can form bacterial flocs, has great application potential in actual wastewater biological treatment projects.

[0074] This experiment also investigated the degradation of amide heterocyclic compounds PMP by Rhodococcus R25 under high salt conditions. This experiment was done by adding PMP mother solution into a 250 mL conical flask (containing 100 mL of sterile inorganic salt nitrogen-free culture medium) to a final concentration of 150 mg / L, with an initial inoculation OD of 0.05 and constant temperature shaking culture at 30°C. Each treatment was carried out in parallel in 3 groups to investigate the degradation and ammoniation of PMP by Rhodococcus R25 under low and high salt conditions, and verification experiments were carried out at two different salinities of 0.1% and 3%. Fig. 9 As shown in the data, Rhodococcus sp. R25 could completely degrade PMP (150 mg / L) within 3 days at both salinities, even though it was partially inhibited by the high salt environment in the initial stage of degradation at 3% salinity. At the same time, the release rates of NH3-N were as high as 95.11% and 96.64% at the two salinities. These results fully demonstrated that Rhodococcus sp. R25 has a wide spectrum of amide compound substrates and can effectively cope with the adverse effects of high salt environment, and has great potential for bioaugmentation applications.

[0075] The above contents are further detailed descriptions of the present invention in combination with specific preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For ordinary technicians in the technical field to which the present invention belongs, several simple deductions or substitutions can be made without departing from the concept of the present invention, which should be regarded as falling within the protection scope of the present invention.

Claims

1. A salt-tolerant amide organic matter-degrading bacterium, characterized in that: It includes: Arthrobacter Paenarthrobacter sp.) P24, deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, deposit date: July 20, 2022; deposit number: CGMCC No .25361; and / or Nitratireductor indicus N30, deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, deposit date: July 20, 2022; deposit number: CGMCC No .25360; and / or Rhodococcus ( Rhodococcus sp.) R25, deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit date is July 20, 2022; the deposit number is CGMCC NO. 25362.

2. The salt-tolerant amide organic matter-degrading bacteria according to claim 1, characterized in that: Its composition and volume ratio are Arthrobacteroids ( Paenarthrobacter sp.)P24 20-60%, Nitratireductor indicus N30 20-40%, Rhodococcus ( Rhodococcus sp.) R25 20-40%.

3. The salt-tolerant amide organic matter-degrading bacteria according to claim 1 or 2, characterized in that: The amide organic matter includes at least one of AAA and PMP.

4. The use of the salt-tolerant amide organic matter degrading bacteria as claimed in claim 1, characterized in that: It is used for the degradation of AAA / PMP in amide-containing organic wastewater.

5. The use of the salt-tolerant amide organic matter degrading bacteria according to claim 4, characterized in that: The degradation environment of the salt-tolerant amide organic matter degrading bacteria is: in an environment with a salt content of 0.1-5%, a temperature of 15-40° C., an initial pH of 5-9, and an aerobic condition.

6. The use of the salt-tolerant amide organic matter degrading bacteria according to claim 4, characterized in that: The dosage of the salt-tolerant amide organic matter degrading bacteria is 0.1 to 1 wt‰ of the organic wastewater.

7. The use of the salt-tolerant amide organic matter degrading bacteria as claimed in claim 1, characterized in that: The salt-tolerant amide organic matter degrading bacteria are used for the degradation of amide organic matter AAA / PMP remaining in sewage, bottom sediment and soil.

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