A strain of Bacillus glutamicum and its bacterial agent and its application in degrading nitrogenous compounds

Abstract

The present invention relates to a strain of Bacillus glutamicum and its bacterial agent and application in degrading nitrogen-containing compounds, belonging to the technical field of microorganisms. Glutamicibacter arilaitensis )XF1‑2, the strain is deposited in the General Microbiology Center of China Microbial Culture Collection Committee, with a deposit number of CGMCC No.32254, a deposit date of October 17, 2024, and an address of the depository institution: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The research glutamicum Bacillus XF1‑2 provided by the present invention has a strong degradation ability for nitrates, nitrites and ammonium salts in sewage, and an addition amount of 0.05‰ can meet the use requirements of sewage treatment. Compared with the existing research glutamicum Bacillus, the ammonia nitrogen removal rate is more than 3.5 times higher.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a strain of Bacillus glutamicum and a bacterial agent thereof and application in degrading nitrogen-containing compounds. Background Art

[0002] At present, the increase in the emission of nitrogen-containing compounds has led to the increasingly serious nitrogen pollution problem in surface water bodies. Ecosystem damage problems such as eutrophication of water bodies are all caused by excessive emission of ammonia nitrogen. Nitrogen pollution poses a serious threat to human health. Therefore, the treatment of nitrogen-containing wastewater has become an important research direction in the field of environmental protection.

[0003] Industrial wastewater is an important source of water pollution, with the characteristics of large volume, wide area, complex composition, high toxicity, difficult to purify, and difficult to treat. Industrial wastewater is mainly concentrated in the petrochemical, oil refining, printing and dyeing industries, among which the petrochemical industry is a pillar industry and also a key industry in the field of water pollution control.

[0004] Nitrogen compounds are essential nutrients for the growth and reproduction of microorganisms, but excessive nitrogen content in natural water bodies can easily lead to eutrophication of water bodies, resulting in the crazy growth of lower organisms (such as cyanobacteria) in the water body, and eventually causing the water body to turn black and smelly. In the process of wastewater treatment, excessive concentrations of free ammonia will have a toxic effect on microorganisms, resulting in reduced microbial activity and affecting the system treatment effect. At the same time, inorganic salts in wastewater play a huge role in maintaining the osmotic pressure of microbial cells and promoting enzyme reactions, but when the content is too high, it will have a serious impact on microorganisms.

[0005] The industrial application of wastewater denitrification is mainly divided into three categories: physical method, chemical method and microbial method. Among them, biological denitrification has the advantages of economy, efficiency and easy operation compared with physical and chemical methods, and thus has become the research focus in the field of denitrification. Many scholars have proposed the theory of bio-augmentation technology, which increases the number of microbial strains by adding specific ammonia nitrogen biodegradation strains, accelerates the recovery of the system, and improves the ammonia nitrogen removal efficiency of the system. Therefore, the development of efficient ammonia nitrogen degrading bacteria is of great significance for the enhanced removal of ammonia nitrogen in wastewater, controlling nitrogen pollution, and alleviating eutrophication of water bodies.

[0006] Research Society of Bacillus glutamicum Glutamicibacter arilaitensis) As a new type of aerobic denitrifying bacteria, relevant research reports have been published in relevant literature. For example, the document Efficient hydroxylamine removal through heterotrophic nitrification by novel bacterium Glutamicibacter arilaitensis EM-H8 (Chen, M., Ding, C., He, T., Zhang, M. and Wu, Q. 2021. Chemosphere 288 (Pt 1), 132475) discloses the research on the removal of hydroxylamine by Glutamicibacter arilaitensis. The experiment lasted for 91 hours under the conditions of carbon-nitrogen ratio of 25, carbon source of glucose, temperature of 20 ° C, and shaker speed of 150 rpm, and the initial nitrogen removal rate was 26.86%. However, it can only remove hydroxylamine in a specific environment, and its removal effect in sewage environment and on other inorganic nitrogen-containing compounds is still limited. Summary of the invention

[0007] In view of the problem that the existing Bacillus glutamicum strains are not effective in removing nitrogen-containing inorganic substances in sewage and require additional addition of carbon sources, the present invention provides a strain of Bacillus glutamicum strains and its bacterial agent and application in degrading nitrogen-containing compounds to solve the above problems.

[0008] In a first aspect, the present invention provides a strain of Bacillus glutamicum, wherein the strain is Bacillus glutamicum ( Glutamicibacter arilaitensis ) XF1-2, the strain has been deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No.32254, the deposit date is October 17, 2024, and the address of the deposit institution is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0009] In a second aspect, the present invention provides a bacterial agent containing Bacillus glutamicum, wherein the bacterial agent is a liquid bacterial agent.

[0010] Furthermore, the number of viable bacteria of Bacillus glutamicum in the liquid bacterial agent is ≥ 2×10 9 CFU / ml.

[0011] A method for preparing the above-mentioned liquid bacterial agent comprises the following steps:

[0012] Liquid culture medium, including (NH 4 ) 2 SO 4 1g / L, K 2 HPO 4 5.45g / L, KH2 PO 4 0.4g / L, MgSO 4 7H 2 O0.18g / L, CaCl 2 ·2H 2 O 0.02g / L, FeSO 4 7H 2 O 0.01g / L, CuSO 4 ·5H 2 O 0.2mg / L, NaH 2 PO 4 0.46g / L, NaCO 3 60g / L, pH=7.3~7.5; sterilize at 121℃ for 30min, cool to 30℃; then connect the research glutamicum XF1-2 to the fermentation tank under flame protection, with an inoculation amount of 2%, a rotation speed of 180rpm, an initial ventilation ratio of 0.5:1, and when the dissolved oxygen drops to 30%, gradually increase the air flow to a ventilation ratio of 1:1, and the fermentation cycle is 16~20h.

[0013] Furthermore, 0.5% potassium dihydrogen phosphate and 1% anhydrous calcium chloride were added to the liquid bacterial agent as protective agents.

[0014] In a third aspect, the present invention provides an application of Bacillus glutamicum in degrading nitrogen-containing compounds, wherein the nitrogen-containing compound is nitrite.

[0015] In a fourth aspect, the present invention provides a method for using Bacillus glutamicum in degrading nitrogen-containing compounds, wherein the nitrogen-containing compounds are nitrates.

[0016] In a fifth aspect, the present invention provides a method for studying the use of Bacillus glutamicum in degrading nitrogen-containing compounds, wherein the nitrogen-containing compounds are ammonium salts.

[0017] The beneficial effects of the present invention are:

[0018] The research glutamicum XF1-2 provided by the present invention has a strong degradation ability for nitrogen-containing inorganic substances in sewage, such as ammonium salts, nitrates and nitrites, and an addition amount of 0.05‰ can meet the use requirements of sewage treatment. Compared with the existing research glutamicum, the ammonia nitrogen removal rate is more than 3.5 times higher. At the same time, the research glutamicum XF1-2 provided by the present invention is a heterotrophic and autotrophic bacterium, which is simple to ferment, does not require activation when adding, does not require additional addition of carbon source, and a single strain can achieve nitrification-denitrification, reducing the complexity of multi-strain addition operation, reducing the workload of the anaerobic tower, and achieving sludge reduction. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0020] Figure 1 It is a graph showing ammonia nitrogen degradation by different strains in Example 1 of the present invention.

[0021] Figure 2 This is a growth diagram of the glutamicum bacteria XF1-2 in LB medium in Example 1 of the present invention.

[0022] Figure 3 This is the Gram staining picture of the glutamicum bacteria XF1-2 studied in Example 1 of the present invention.

[0023] Figure 4 This is a graph showing the degradation of nitrite by the glutamicum bacteria XF1-2 in Example 2 of the present invention.

[0024] Figure 5 This is a graph showing the degradation of nitrate by the glutamicum bacteria XF1-2 in Example 2 of the present invention.

[0025] Figure 6 This is a diagram of the simulated sewage experiment of the glutamicum Bacillus XF1-2 in Example 3 of the present invention.

[0026] Figure 7 It is a growth curve of the glutamicum bacteria XF1-2 in LB medium in Example 4 of the present invention. DETAILED DESCRIPTION

[0027] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0028] Example 1

[0029] Isolation, Screening and Identification of Bacillus glutamicum XF1-2

[0030] 1. Sampling

[0031] Biochemical pond wastewater is collected in Zibo City, Shandong Province on June 8, 2024.

[0032] 2. Separation

[0033] Centrifuge at 8000 rpm for 10 min in a high-speed centrifuge, take the bacterial sludge, wash with sterilized 0.8% saline, centrifuge for 3 times, take 1 ml of the bacterial sludge, and spread it on a sterilized Luria-Bertani solid medium (10 g of tryptone, 5 g of yeast extract, 10 g of NaCl, 15-20 g of agar, distilled water to 1000 ml, pH = about 7.0) for gradient dilution, place in a constant temperature incubator at 30°C for about 24 h, screen out 9 strains at 10-7 and 10-8 (labeled as XF1-1, XF1-2, XF1-3, XF1-4, XF1-5, XF1-6, XF1-7, XF1-8 and XF1-9 respectively), pick single colonies and store them in 50% sterile glycerol tubes at -80°C.

[0034] 3. Selection of strains that degrade nitrogenous compounds

[0035] The 9 strains stored in the glycerol tubes were streaked onto sterile plates of Luria-Bertani solid culture medium, placed in a 30°C constant temperature incubator for 18-24 hours, and single colonies were isolated (the grown plates were placed in a 4°C refrigerator for refrigeration).

[0036] Ammonia nitrogen test medium is: (NH 4 ) 2 SO 4 0.472g, sodium succinate 4.052g, 50ml of Vickers salt solution, add water to dissolve, add distilled water to 1L, and adjust the pH to 7.0. Wherein Vickers salt solution (g / L): K 2 HPO 4 5.0 g, FeSO 4 7H 2 O 0.05 g, NaCl 2.5 g, MgSO 4 7H 2 O 2.5g, MnSO 4 ·4H 2 O 0.05g. Prepare 9 copies of ammonia nitrogen test culture medium, inoculate 1 ring of activated single colony respectively, place in a 30℃ constant temperature shaker, set the speed to 200rpm, record the initial ammonia nitrogen value and the ammonia nitrogen value every 4h thereafter (equipment model: GNST-900, manufacturer: Henan Suijing Environmental Protection Technology Co., Ltd.), the ammonia nitrogen degradation curve of the strain is as follows Figure 1 Finally, a strain XF1-2 with the fastest degradation rate per unit time was screened out. In 20 hours, the ammonia nitrogen content of the test culture medium was reduced from 150.4 mg / L to below 4.6 mg / L, and the ammonia nitrogen removal rate reached 96.94%, indicating that this strain has the function of ammonia oxidizing bacteria.

[0037] 4. Identification

[0038] The bacteria are club-shaped. The colony morphology on LB plate is light yellow with regular edges, smooth and slightly convex surface. Figure 2 Gram staining microscopy (equipment model: Nikon Ei100, manufacturer: Nanjing Jiangnan Yongxin Optics Co., Ltd.) showed positive results, see Figure 3 On July 13, 2024, the test tube slant was sent to Beijing Qingke Biotechnology Co., Ltd. for identification, and the gene sequence of strain XF1-2 was obtained as shown in SEQ ID NO.1. The identification result was Bacillus glutamicum Glutamicibacter arilaitensis . The glutamicum XF1-2 of the research institute is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No.32254, the deposit date is October 17, 2024, and the address of the deposit institution is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0039] Example 2

[0040] Nitrite and Nitrate Degradation Test

[0041] 1. Nitrite degradation test

[0042] High Nitrite Nitrogen Test Medium: Na 2 HPO 4 5g, KH 2 PO 4 1g, MgSO 4 0.1g, C 4 H 4 Na 2 O 4 6g, NaNO 2 0.08g, trace element solution 2mL, trace element solution: EDTA 50g, ZnSO 4 2.2 g, CaCl 2 ·2H 2 O 5.5 g, MnSO 4 ·4H 2 O 5g, FeSO 4 7H 2 O 5g, (NH 4 ) 6 Mo 7 O 24 ·4H 2 O 1.1g, CuSO 4 ·5H 2 O 1.5 g, CoCl 2 6H 2O 1.6g, add distilled water to 1.0 L, pH 6.0. After the above culture medium is prepared, adjust the pH to 7.9, dispense the liquid culture medium, sterilize at 121℃ for 20min, and the pH after sterilization is 7.86. Select a single colony containing the research association glutamicum XF1-2 to conduct a parallel experiment in a high nitrite nitrogen liquid test culture medium, place it in a 30℃ constant temperature shaker, set the speed to 200rpm, record the initial value and test the nitrite nitrogen content every 4h (equipment model: GNST-900, manufacturer: Henan Suijing Environmental Protection Technology Co., Ltd.), the results are as follows Figure 4 The glutamicum XF1-2 in the high nitrite medium recovered slightly at the beginning, which may be because the target bacteria just began to adapt to the new environment, resulting in the death of some bacteria. After 4 hours, NO 2 -N began to decrease steadily, and NO 2 -N dropped from a peak of 62mg / L to 0mg / L, NO 2 -N degradation rate is 100%, and the average degradation rate is 2.2mg / L / h, which shows that the glutamicum XF1-2 can oxidize nitrite and has denitrification effect. After the culture is completed, the nitrate nitrogen content in the test culture medium is detected to be 0mg, indicating that the nitrate nitrogen produced is used by denitrification in the biochemical process.

[0043] 2. Nitrate degradation test

[0044] Nitrate nitrogen test medium: CH 3 COONa 12g, KNO 3 0.1~0.2g, K 2 HPO 4 ·3H 2 O 4g, NaCl 4g, MgSO 4 7H 2 O 10mg, MnSO 4 ·H 2 O 10mg, ZnSO 4 7H 2 O 10mg, H 3 BO 3 10mg, FeSO 4 7H 2 O 10mg / L, CaCl 2 20mg, dissolved and diluted to 1000ml distilled water, pH = 7.5, sterilized at 121℃ for 20min, pH = 7.86 after sterilization. Single colonies of glutamicum XF1-2 were selected and added to nitrate nitrogen liquid test medium for parallel experiments, in which group A was added with KNO 3 0.1g, group B plus KNO 30.2g, placed in a 30℃ constant temperature shaker, set the speed to 200rpm, record the initial value and test the nitrate nitrogen content every 4h. The results are as follows Figure 5 As shown in the figure, no matter in the case of low or high concentration of nitrate nitrogen, glutamicum XF1-2 can degrade it to 0, NO 3 -N has an average degradation rate of 3.83 mg / L / h, which is faster than nitrite nitrogen degradation, indicating that nitrite is slightly more toxic than nitrate. The research results show that Bacillus glutamicum XF1-2 can use nitrate nitrogen as a nitrogen source and has denitrification ability.

[0045] Example 3

[0046] Artificial wastewater simulation experiment

[0047] Preparation (NH 4 ) 2 SO 4 0.19g, KNO 3 0.1g, sodium citrate 4.08g, MgSO 4 7H 2 O 0.05g, K 2 HPO 4 ·3H 2 O 0.2 g, NaCl 0.12 g, MnSO 4 ·H 2 O 0.01g, FeSO 4 7H 2 O 0.01g, dissolve the above culture medium and make it up to 1000ml distilled water, pH=7.5, and do not sterilize it. Inoculate the suspension of Bacillus glutamicum XF1-2 cultured in LB sterile culture, inoculate it with 1%, place it in a 30℃ constant temperature shaker, set the speed to 200rpm, record the initial value, and test the ammonia nitrogen, nitrite nitrogen, and nitrate nitrogen values ​​every 4h and record them. The data results are as follows Figure 6 As shown. In 20 hours, all nitrogen compounds dropped to nearly 0 mg, and the effluent met the standard. During the nitrification process, ammonia nitrogen was consumed first. After 16 hours of ammonia nitrogen consumption, nitrate nitrogen continued to be consumed. In theory, in the nitrification reaction, an intermediate product, nitrite nitrogen, would be produced, but it was not detected throughout the process, indicating that with the nitrification-denitrification process, the intermediate product was directly used by denitrification. This experiment also proves that XF1-2 performs nitrification and denitrification simultaneously in sewage, and uses nitrogen-containing compounds to synthesize its own biomass.

[0048] Example 4

[0049] Preparation of liquid inoculum containing Bacillus glutamicum XF1-2

[0050] 1. Shake flask culture: prepare LB liquid culture medium (tryptone 10g, yeast extract 5g, NaCl 10g, agar 15-20g, distilled water to 1000ml, pH=7.0), dispense into baffled triangular flasks (liquid volume 30%~40%), sterilize and cool for use; inoculate single colonies into the first-level shake flasks in a sterile manner on a clean bench, culture at 200rpm, 30℃ for about 16h, then transfer to the second-level LB culture medium for expansion, culture at 200rpm, 30℃ for 10~12h, and then transfer to the fermenter. The growth curve of Bacillus glutamicum XF1-2 in LB was studied, such as Figure 7 As shown, the OD increased from 0.05 to 2.43 in 16 hours, and the number of viable bacteria detected was 2.16 billion / ml.

[0051] 2. Liquid inoculant preparation: sterilize the fermentation tank and prepare liquid culture medium, including (NH 4 ) 2 SO 4 1g / L, K 2 HPO 4 5.45g / L, KH 2 PO 4 0.4g / L, MgSO 4 7H 2 O 0.18g / L, CaCl 2 ·2H 2 O 0.02g / L, FeSO 4 7H 2 O0.01g / L,CuSO 4 ·5H 2 O 0.2mg / L, NaH 2 PO 4 0.46g / L, NaCO 3 60g / L, pH=7.3~7.5; sterilize at 121℃ for 30min, cool to 30℃; connect the secondary shake flask to the fermenter (inoculation amount 2%) under flame protection, rotate at 180rpm, initial ventilation ratio 0.5:1, wait until the dissolved oxygen drops to 30%, gradually increase the ventilation ratio to 1:1; pH drops to about 5.8, cycle 16~20h, viable bacteria count>2×10 9 CFU / ml fermentation was over, 0.5% potassium dihydrogen phosphate and 1% anhydrous calcium chloride were added as protective agents to obtain a liquid bacterial agent containing the research group glutamicum XF1-2.

[0052] Example 5

[0053] On-site oily wastewater experiment

[0054] Experiment 1: Take a sewage sample from a sedimentation tank of Shanghai Gaoqiao Petrochemical, first adjust the pH to 7.50, and then inoculate the sewage sample with the fermentation liquid of Bacillus glutamicum XF1-2 according to different inoculation amounts. Control group: sewage stock solution; Group A: sewage stock solution + 0.05‰ fermentation liquid of Bacillus glutamicum XF1-2; Group B: sewage stock solution + 0.1‰ fermentation liquid of Bacillus glutamicum XF1-2; Group C: sewage stock solution + 0.3‰ fermentation liquid of Bacillus glutamicum XF1-2; Group D: sewage stock solution + 0.5‰ fermentation liquid of Bacillus glutamicum XF1-2. After the fermentation liquid is shaken, the initial value is detected, and the aeration culture is carried out in a constant temperature shaker at 180rpm and 30℃. The experimental data of each group is detected and recorded every 8 hours. The specific results are shown in Table 1 below:

[0055] Table 1- Effect of different inoculation amounts on reducing ammonia nitrogen in sewage

[0056]

[0057] From the results in Table 1, it can be seen that the initial value of ammonia nitrogen increases with the increase of inoculation amount, because the bacterial solution itself has (NH 4 ) 2 SO 4 Residual and NH 4 + The experiment showed that the degradation rate of ammonia nitrogen in the four groups with fermentation liquid added was faster than that in the blank control group, and the degradation rate increased with the increase of fermentation liquid added. The ammonia nitrogen value of group D was 1.8 mg / L after 24 hours of aeration, which had reached the emission standard (ammonia nitrogen value < 4 mg / L). The ammonia nitrogen value of group A also dropped to 0.8 mg / L at 40 hours, which also reached the emission standard. In view of the cost considerations in the actual application process, as well as the experience of the company's dosage ratio and frequency, the dosage of group A can meet the on-site needs.

[0058] At the end of the experiment, the nitrite nitrogen and nitrate nitrogen in the sewage samples were tested, both of which were 0 mg / L. The actual sewage treatment results were consistent with those of the simulated sewage, laying the foundation for industrialization.

[0059] Experiment 2: Field experiment: Aeration tank wastewater experiment of the second workshop of Sinopec Qilu Branch Refinery Water Service, with an inlet flow rate of 600m 3 / h. Dosing scheme: 90 kg of the liquid bacterial agent containing Bacillus glutamicum XF1-2 prepared in Example 4 was added at 0 h, and 30 kg of the liquid bacterial agent was added every 24 h thereafter. The sampling test results are shown in Table 2.

[0060] Table 2- Effect of different inoculation amounts on reducing ammonia nitrogen in sewage

[0061]

[0062] As can be seen from Table 2, after adding the liquid bacterial agent containing the research institute's glutamicum Bacillus XF1-2, the ammonia nitrogen removal effect was obvious, and the ammonia nitrogen value began to decrease. It also shows that the research institute's glutamicum Bacillus XF1-2 can adapt to the petrochemical site environment. After 30 hours of action, the ammonia nitrogen can reach the national emission standard of ≤4mg / L, and after 36 hours it reaches ≤1mg / L. After 36 hours, it slows down and tends to stabilize, and the ammonia nitrogen effluent meets the standard.

[0063] Although the present invention has been described in detail with reference to the accompanying drawings and in combination with the preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and these modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily think of changes or substitutions within the technical scope disclosed by the present invention, and these shall be within the scope of protection of the present invention.

Claims

1. A strain of Bacillus glutamicum, characterized in that: The research group glutamicum is the research group glutamicum ( Glutamicibacter arilaitensis ) XF1-2, the strain is deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, the deposit number is CGMCC No.32254, the deposit date is October 17, 2024, and the address of the deposit institution is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. The researched Bacillus glutamicum according to claim 1, characterized in that The 16S rDNA sequence of the glutamicum bacteria is shown in SEQ ID NO.

1.

3. A bacterial agent containing the Bacillus glutamicum of claim 1, characterized in that: The bacterial agent is a liquid bacterial agent.

4. The bacterial agent according to claim 3, characterized in that The number of viable bacteria of the researched glutamicum in the liquid bacterial agent is ≥ 2×10 9 CFU / ml.

5. The bacterial agent according to claim 3, characterized in that The preparation method of the liquid bacterial agent comprises the following steps: preparing a liquid culture medium, including (NH4)2SO41g / L, K2HPO45.45g / L, KH2PO40.4g / L, MgSO4·7H2O0.18g / L, CaCl2·2H2O0.02g / L, FeSO4·7H2O0.01g / L, CuSO4·5H2O0.2mg / L, NaH2PO40.46g / L, NaCO360g / L, pH=7.3-7.5; sterilizing at 121°C for 30min, cooling to 30°C; then connecting the research glutamicum XF1-2 to a fermentation tank under flame protection, with an inoculation amount of 2%, a rotation speed of 180rpm, an initial ventilation ratio of 0.5:1, and gradually increasing the air flow to a ventilation ratio of 1:1 when the dissolved oxygen drops to 30%, and a fermentation cycle of 16-20h.

6. The bacterial agent according to claim 3, characterized in that 0.5% potassium dihydrogen phosphate and 1% anhydrous calcium chloride were added to the liquid bacterial agent as protective agents.

7. A use of the research bacillus glutamicum in degrading nitrogen-containing compounds as claimed in claim 1, characterized in that: Nitrogen-containing compounds are nitrites.

8. A use of the research bacillus glutamicum in degrading nitrogen-containing compounds as claimed in claim 1, characterized in that: Nitrogen-containing compounds are nitrates.

9. A use of the research bacillus glutamicum in degrading nitrogen-containing compounds as claimed in claim 1, characterized in that: The nitrogen-containing compound is an ammonium salt.

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