Acinetobacter severii DE-5 and its application

Abstract

The present invention relates to the technical field of microorganisms, and more particularly to a strain of Acinetobacter severii DE-5 and its applications. The strain was deposited in the Wuhan Microbiology Collection Center on June 19, 2024, with a deposit number of CCTCC NO: M20241302. The strain of the present invention has the ability to degrade organic phosphorus, inorganic phosphorus, and cellulose, providing basic conditions for in-situ remediation of pesticide residues in soil and improving soil nutrition.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganisms, and more particularly to Acinetobacter severii DE-5 and applications thereof. Background Art

[0002] Phosphorus is one of the essential nutrients for plant growth, second only to nitrogen in importance, accounting for about 0.2% of the dry weight of plants. It is a major component involved in energy metabolism, nucleic acid and cell synthesis, and the regulation of some enzymes. It plays an important role in promoting plant growth, development and metabolism, and is an irreplaceable component in the ecosystem.

[0003] Phosphorus in the soil exists in two forms: inorganic and organic compounds. However, only a small amount of phosphorus can be utilized by plants, becoming a limiting factor affecting plant growth. Most of the phosphorus in chemical phosphorus fertilizers applied to the soil will remain in the soil, causing soil and water pollution. Among them, ethoprophos, an organophosphorus insecticide widely used in agriculture, is highly toxic. Its molecular structure is C8H 19 O2PS2 is a colorless liquid or white solid that is easily soluble in organic solvents. Ethoprophos is widely used in the production of various crops, including fruit trees and vegetables, due to its effectiveness in preventing and controlling common nematode diseases in crops. However, due to its high mobility, ethoxyphos residues in the soil pose a potential hazard to soil and groundwater, polluting the surface soil ecosystem and indirectly harming human health. Therefore, reducing the residual levels of organic and inorganic phosphorus, such as ethoxyphos, in the soil has a positive impact on plant growth and the remediation of pesticide-contaminated soil and water.

[0004] There are currently a variety of treatment methods for the problem of residual organic and inorganic phosphorus in the soil, including chemical methods and biological methods. The essence of the chemical method is to change the chemical properties of phosphorus through chemical reactions, thereby reducing its biological activity or converting it into harmless substances. When applied to actual production, not only is the cost high, but the chemical reagents themselves may be harmful to the environment, change the physical properties of the soil, and easily cause secondary pollution. The biological method uses specific microorganisms to convert phosphorus into harmless substances. Biodegradation is a naturally occurring chemical process that uses specific microorganisms to accelerate this process without producing additional intermediates or pollution. In comparison, biodegradation is usually low in cost, easy to operate, and has advantages such as good phosphorus removal and no secondary pollution.

[0005] Currently, the bacterial species reported to have the ability to degrade both organic and inorganic phosphorus, including ethoprophos, include Bacillus badius, Serratia liquefaciens, Klebsiella aerogenes, and Stenotrophomonaspavanii. Summary of the Invention

[0006] To solve the above problems, the present invention provides an Acinetobacter severii DE-5 and its application, so as to enrich the bacterial species capable of degrading organic phosphorus and inorganic phosphorus.

[0007] The present invention is achieved through the following technical solutions:

[0008] The first aspect of the present invention provides a strain of Acinetobacter seifertii and its application, wherein the strain is Acinetobacter seifertii DE-5, which was deposited in the China Center for Type Culture Collection on June 19, 2024, with a deposit number of CCTCC NO: M20241302.

[0009] The second aspect of the present invention provides a bacterial agent, the active ingredient of which is the Acinetobacter severii DE-5 or a culture of Acinetobacter severii DE-5.

[0010] Preferably, the method for preparing the culture of Acinetobacter severiensis DE-5 comprises the following steps: inoculating the Acinetobacter severiensis DE-5 into a culture medium suitable for bacteria, culturing on a shaking table at 20°C to 40°C for 11h to 13h, and removing the bacteria to obtain a culture of Acinetobacter severiensis DE-5.

[0011] The third aspect of the present invention provides a use of the bacterial agent in degrading organic phosphorus, wherein the organic phosphorus is ethoprophos and lecithin.

[0012] A fourth aspect of the present invention provides a use of the bacterial agent in degrading inorganic phosphorus, wherein the inorganic phosphorus is tricalcium phosphate.

[0013] A fifth aspect of the present invention provides a use of the bacterial agent in degrading cellulose.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] (1) The present invention screened out Acinetobacter safferii DE-5, which has the ability to degrade chloranthate, from the Yangtze River to Jichao River. Through continuous screening from a large number of bacterial strains, isolation and purification, identification tests, growth performance, and degradation capacity testing, the selected Acinetobacter safferii exhibited a high chloranthate degradation efficiency. When cultured for 7 days at 35°C and 150 rpm, the strain achieved a degradation efficiency of 70.68% in a 10 mg / L inorganic salt medium and 82.95% in a medium supplemented with 0.1 g / L glucose. This enriched the chloranthate-degrading bacterial pool and provided basic conditions for in-situ remediation of pesticide residues in soil.

[0016] (2) Acinetobacter severii DE-5 also dissolves organic phosphorus, inorganic phosphorus, and cellulose, with the inorganic phosphorus dissolving being the most effective. Therefore, DE-5 can be used to promote the decomposition of organic phosphorus, cellulose, and especially inorganic phosphorus in the soil, thereby improving the soil's nutritional status.

[0017] Biomaterial deposit information

[0018] Acinetobacter seifertii DE-5 was deposited in the China Center for Type Culture Collection on June 19, 2024, with the deposit number CCTCC NO: M20241302, and the deposit address is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province. 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 following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0020] Figure 1 This is a colony morphology diagram of Acinetobacter severii DE-5 strain in an embodiment of the present invention;

[0021] Figure 2 The morphology of Acinetobacter severii DE-5 strain under a staining microscope in an embodiment of the present invention is shown;

[0022] Figure 3 : is a Gram staining image of Acinetobacter severii DE-5 strain in an embodiment of the present invention;

[0023] Figure 4 1 is a homology analysis diagram of Acinetobacter severii DE-5 strain in the embodiment of the present invention;

[0024] Figure 5Graph showing the effect of temperature on the growth of Acinetobacter severii DE-5 in an embodiment of the present invention;

[0025] Figure 6 Graph showing the effect of pH on the growth of Acinetobacter severii DE-5 in an embodiment of the present invention;

[0026] Figure 7 This is a standard curve diagram for the detection of ethoprophos in an embodiment of the present invention;

[0027] Figure 8 This is a graph showing the degradation efficiency of different concentrations of ethoprophos by Acinetobacter severi DE-5 in an embodiment of the present invention;

[0028] Figure 9 This is a peak diagram of the ethoprophos pesticide in an embodiment of the present invention after detection by gas chromatography-mass spectrometry;

[0029] Figure 10 This is a peak graph of Acinetobacter serrata DE-5 against the same concentration of ethoprophos after gas chromatography-mass spectrometry detection in an embodiment of the present invention;

[0030] Figure 11 This is a diagram showing the effect of dissolving organic phosphorus in an embodiment of the present invention;

[0031] Figure 11 Among them, A is the effect of dissolving organic phosphorus in the phosphorus-dissolving circle for 1 day, and B is the effect of dissolving organic phosphorus in the phosphorus-dissolving circle for 3 days;

[0032] Figure 12 This is a diagram showing the effect of dissolving inorganic phosphorus in an embodiment of the present invention;

[0033] Figure 12 Among them, A is the effect of dissolving inorganic phosphorus by dissolving phosphorus circle for 1 day, and B is the effect of dissolving inorganic phosphorus by dissolving phosphorus circle for 3 days;

[0034] Figure 13 This is a diagram showing the effect of dissolving cellulose in an embodiment of the present invention;

[0035] Figure 13 In the figure, A is the effect of dissolving cellulose and phosphorus-dissolving ring for 1 day, and B is the effect of dissolving cellulose and phosphorus-dissolving ring for 3 days;

[0036] Figure 14 This is a diagram showing the measurement results of the dissolved organic phosphorus ring diameter in an embodiment of the present invention;

[0037] Figure 15 This is a diagram showing the measurement results of the dissolved inorganic phosphorus ring diameter in an embodiment of the present invention;

[0038] Figure 16 This is a diagram showing the measurement results of the diameter of the dissolved cellulose ring in an embodiment of the present invention;

[0039] Figure 17This is a standard curve diagram for determining the available phosphorus content in the examples of the present invention. DETAILED DESCRIPTION

[0040] To facilitate understanding of the present invention, the present invention will be described more fully below, along with preferred embodiments thereof. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a more thorough and comprehensive understanding of the disclosure of the present invention.

[0041] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in this specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0042] The beneficial effects of the present invention are described below by means of specific embodiments:

[0043] Example 1, Isolation and purification of Acinetobacter severii DE-5

[0044] The components of the culture medium used in this example are:

[0045] Beef extract peptone medium: 3 g beef extract, 5 g NaCl, 10 g peptone, 1000 mL ultrapure water; pH 6.8, divided into conical flasks, and sterilized at 121°C.

[0046] Inorganic salt solid culture medium: Na2HPO4 6.34 g, KH2PO4 1.33 g, (NH4)2SO4 1 g, MgSO4·7H2O 0.2 g, FeSO4 0.001 g, CaCl2 0.04 g, deionized water 1000 mL; pH 6.8, divided into conical flasks, and sterilized at 121°C.

[0047] Glucose-peptone medium: 5 g glucose, 5 g NaCl, 5 g peptone, 1000 mL ultrapure water; pH 6.8, divided into conical flasks, and sterilized at 121°C.

[0048] Weigh 5g of soil samples from along the Yinjiang-Jichao River, place them in a centrifuge tube filled with 10mL of sterile water, and shake them thoroughly; after standing, take the supernatant and place it in 100mL of beef extract peptone medium, and culture it in a shaker at 30℃ and 150r / min for 1 day; use an inoculation loop to dip the culture solution, streak it on a beef extract peptone solid medium plate with a concentration of 30mg / L of chloranthate, and place it in a 30℃ biochemical incubator for 2 days; take the grown colonies and inoculate them on beef extract peptone solid medium with a concentration of 60mg / L and 120mg / L of chloranthate in turn; then take the acclimated colonies and inoculate them on inorganic salt solid medium with a concentration of 240mg / L and 500mg / L of chloranthate in turn, and culture them in a 30℃ biochemical incubator for 10 days; take the colony grown at the highest pesticide concentration to amplify the bacterial solution, and spread it on the plate to screen for single strains. The bacterial solution was diluted to 10 with sterile water in sequence. -5 , 10 -6 , 10 -7 , 10 -8 , 10 -9 , spread the plate, screen single colonies, preserve the seeds in glycerol, and store in a -60℃ ultra-low temperature refrigerator.

[0049] Example 2, Identification of Acinetobacter severii DE-5

[0050] (1) Morphological identification of strains: After the isolated strains are amplified, they are streaked on solid culture medium and incubated at 30°C for 1 day to observe the basic characteristics of the strains, such as shape, edge, and color. The colonies are taken and Gram staining is used to identify the type. If the strain is Gram-negative, Figure 3 shown.

[0051] like Figure 1 As shown in the figure, the colony morphology of Acinetobacter severii DE-5 in the embodiment of the present invention is shown. The strain appears as white, rod-shaped colonies on the plate with neat edges, round, smooth surface, and upward protrusions. Figure 2 shown.

[0052] (2) Physiological and biochemical identification of strains:

[0053] Acinetobacter severii DE-5 strain was subjected to MR test, VP test, gelatin hydrolysis test, starch hydrolysis test, nitrate reduction test, and catalase test. Physiological and biochemical identification tests were performed on each strain, and the results are shown in Table 1.

[0054] Table 1 Physiological and biochemical identification

[0055]

[0056] Note: "+" indicates positive, "-" indicates negative

[0057] (3) 16S rDNA Sequencing: 16S rDNA was used as the target for sequencing analysis of microorganisms to determine bacterial species. The strain gene was extracted using a bacterial genomic DNA extraction kit. Using this as a template, PCR was performed to amplify the 16S rDNA sequence of Acinetobacter severii DE-5.

[0058] The primer sequences are as follows:

[0059] The forward primer for PCR amplification was B341F: 5′-CCTACGGGNGGCWGCAG-3′.

[0060] The reverse primer was B785R: 5′-GACTACHVGGGTATCTAAT-3′.

[0061] The PCR reaction program was as follows: pre-denaturation at 95°C for 3 min, denaturation at 95°C for 30 s, 25 cycles, annealing at 54°C for 30 s, extension at 72°C for 30 s, and final extension at 72°C for 5 min.

[0062] Sequencing of amplified products:

[0063] The PCR products were subjected to 2% agarose gel electrophoresis and sent to Jiangsu Sanshu Biotechnology Co., Ltd. for sequencing. The sequencing results were analyzed using the BLAST program in the NCBI database to search for homologous sequences. Figure 4 As shown, the strain had a 100% homology with Acinetobacter seifertii, so it was named Acinetobacter seiferti DE-5.

[0064] Example 3. Growth characteristics of Acinetobacter severii DE-5

[0065] (1) Growth of strains at different temperatures: Use an inoculating loop to dip the overnight culture solution into beef extract peptone medium with a pH of 6.8, and culture it at 20°C, 25°C, 30°C, 35°C, and 40°C, respectively, in a shaking incubator at 150 rpm for 12 h. The absorbance of the culture solution at a wavelength of 600 nm was measured using an ultraviolet spectrophotometer, abbreviated as OD. 600 .

[0066] (2) Growth of strains at different pH values: The pH values ​​of beef extract peptone medium were adjusted to 5, 6, 7, 8, and 9, respectively. An inoculating loop was used to dip the overnight culture solution into the medium at different pH values. The culture was shaken at 30°C and 150 rpm, and the absorbance at 600 nm was measured every 4 h.

[0067] like Figure 5As shown in the figure, the results showed that temperature had a great influence on the growth of Acinetobacter severii DE-5. In the range of 20℃~40℃, the overall growth rate first increased and then stabilized or decreased, and the optimum temperature was 35℃.

[0068] like Figure 6 As shown, the results showed that pH affected the growth concentration of Acinetobacter severii DE-5 and the optimal growth pH was 6-7.

[0069] Example 4, establishment and verification of the determination method of ethoprophos

[0070] (1) Gas chromatography-mass spectrometry conditions: A GCMS-TQ8040 triple quadrupole gas chromatography-mass spectrometer was used. The chromatographic column was an SH-Rxi-17Sil MS column with a length of 30 m, an inner diameter of 0.25 mm, and a polar column with a filler particle size of 0.25 μm. Methods: The inlet temperature was 250°C, the interface temperature was 280°C, the ion source temperature was 240°C, the carrier gas was 99.999% pure helium, the flow rate was 1.97 mL / min, the injection volume was 1 μL, and the selected ion scan mode was used. The temperature was programmed to rise to 65°C for 1 min, then to 130°C at 20°C / min, then to 280°C at 10°C / min and held for 10 min, and finally to 300°C at 10°C / min and held for 10 min.

[0071] (2) Preparation of standard curve: dilute the ethoprophos standard with acetonitrile to the concentrations of 2 mg / L, 5 mg / L, 10 mg / L, 20 mg / L, and 50 mg / L for testing. Figure 7 As shown, the concentration of anthrophos is plotted as the horizontal axis and the corresponding peak area is plotted as the vertical axis, and the following standard curve is obtained: y = 851409x-779987, R 2 =0.99.

[0072] (3) Recovery Determination: Ethoprophos concentrations of 10 mg / L, 20 mg / L, and 50 mg / L were set in an inorganic salt liquid culture medium. No degrading bacteria were inoculated and the culture was shaken at 30°C and 150 rpm for 7 days. Three replicates were set, and the recovery of the method was determined by gas chromatography-mass spectrometry. The results are shown in Table 2. In the inorganic salt culture medium supplemented with different concentrations of ethoxyprophos, the recovery of ethoxyprophos ranged from 95.41% to 105.79%, with a coefficient of variation between 2.76% and 6.16%, meeting the requirements for pesticide residue analysis.

[0073] Table 2 Recovery rate and coefficient of variation of ethoprophos added to culture medium

[0074] Ethoprophos concentration, mg / L Average recovery rate, % Coefficient of variation, % 10 105.78 4.56 20 97.51 2.76 50 95.41 6.16

[0075] Example 5: Determination of the degradation efficiency of ethoprophos by Acinetobacter severi DE-5

[0076] The isolated and purified strain in Example 1 was activated overnight, washed once with PBS at pH 7, and resuspended to adjust the bacterial solution. The absorbance of the bacterial solution at a wavelength of 600 nm was measured by ultraviolet spectrophotometer to be 2.0. The culture medium was then inoculated into fermentation medium with 10 mg / L, 50 mg / L, and 100 mg / L of chloranthropyrin concentrations, and samples were taken at regular intervals to determine the degradation rate of the organophosphorus pesticide. Figure 8 As shown in the figure, the degradation efficiency in a normal inorganic salt medium with a concentration of 100 mg / L was 25.63%, and in a medium supplemented with 0.1 g / L DE-5 glucose, the degradation efficiency reached 45.13%. The degradation efficiency in a normal inorganic salt medium with a concentration of 50 mg / L was 51.25%, and in a medium supplemented with 0.1 g / L glucose, the degradation efficiency reached 60.65%. The degradation efficiency in a normal inorganic salt medium with a concentration of 10 mg / L was 70.68%, and in a medium supplemented with 0.1 g / L glucose, the degradation efficiency reached 82.95%. Figure 8 It can be seen that DE-5 has a high degradation ability of ethoprophos, especially the degradation efficiency at a low concentration of 10 mg / L. The degradation rate of the strain reached 82.95% on the 7th day.

[0077] In addition, the pesticide ethoprophos was tested by gas chromatography-mass spectrometry, i.e. GC-MS, and the test results were as follows: Figure 9 As shown, the Acinetobacter severii DE-5 strain isolated and purified in Example 1 was inoculated into culture media containing 10 mg / L, 20 mg / L, and 50 mg / L of ethoprophos, respectively, and then the ethoprophos content was detected by gas chromatography-mass spectrometry. The results are shown in FIG. Figure 10 As shown in the figure, only 10 μg of ethoxychlor remained after degradation, which is below the detection limit. Therefore, this bacterium has good application value in detoxifying ethoxychlor and reducing environmental pollution.

[0078] Example 6: Determination of the Ability of Acinetobacter severii DE-5 to Solubilize Inorganic and Organic Phosphorus

[0079] Gram stain, phosphorus standard solution, ascorbic acid solution, molybdate solution, molybdenum antimony anti-color developer, agarose, DNA extraction kit, glycerol, 75% sulfuric acid solution.

[0080] The culture medium components used in this example are:

[0081] Inorganic phosphorus medium: glucose 10.0 g, ammonium sulfate 0.5 g, yeast extract powder 0.5 g, sodium chloride 0.3 g, potassium chloride 0.3 g, magnesium sulfate 0.3 g, ferrous sulfate 0.03 g, manganese sulfate 0.03 g, tricalcium phosphate 5.0 g, deionized water 1000 mL; pH 7.0.

[0082] Egg yolk agar medium: 15.0 g peptone, 3.0 g beef powder, 5.0 g sodium chloride, 15.0 g glucose, 50% egg yolk solution, 15 g agar, 1000 mL deionized water; pH 7.3.

[0083] Cellulose Congo red medium: sodium nitrate 1.0 g, disodium hydrogen phosphate 1.2 g, potassium dihydrogen phosphate 0.9 g, magnesium sulfate 0.5 g, potassium chloride 0.5 g, yeast extract powder 0.5 g, acid hydrolyzed casein 0.5 g, Congo red 0.2 g, cellulose powder 5.0 g, deionized water 1000 mL, agar 15.0 g; pH 7.0.

[0084] Identification of the strain's phosphate-solubilizing ability: Lecithin was used as the only organic phosphorus source and Ca3(PO4)2 was used as the only inorganic phosphorus source.

[0085] Qualitative determination of dissolved phosphorus:

[0086] Three wells were punched on the egg yolk agar medium using a hole puncher, and 30 μL of the bacterial solution of Acinetobacter severii DE-5 isolated and purified in Example 1 was inoculated into each well. Three parallel experiments were set up, labeled as sample 1, sample 2, and sample 3. The wells were cultured at 28°C for 1 to 3 days, and the diameter of the dissolution zone was measured at 1 day and 3 days respectively. Figure 11 As shown in the figure, A is the effect of the dissolved organic phosphorus dissolving circle for 1 day, and B is the effect of the dissolved organic phosphorus dissolving circle for 3 days. The measurement results of the dissolved organic phosphorus circle diameter are as follows Figure 14 As shown, the diameter of the dissolution zone of dissolved organic phosphorus measured on day 1 reached 7.35 mm to 10.19 mm, and the diameter of the dissolution zone of dissolved organic phosphorus measured on day 3 reached 7.69 mm to 10.89 mm.

[0087] Use a hole puncher to make three holes in the inorganic phosphorus culture medium, and inoculate 30 μL of the bacterial solution of Acinetobacter severii DE-5 isolated and purified in Example 1. Set up three parallel experiments, labeled as sample 4, sample 5 and sample 6, and culture at 28°C for 1 to 3 days. The diameter of the dissolution zone is measured at 1 day and 3 days respectively. Figure 12 As shown in the figure, A is the effect of dissolving inorganic phosphorus in the phosphorus circle for 1 day, and B is the effect of dissolving inorganic phosphorus in the phosphorus circle for 3 days. The measurement results of the diameter of the dissolved inorganic phosphorus circle are as follows Figure 15 As shown, the diameter of the dissolution zone of dissolved inorganic phosphorus measured on day 1 reached 8.2 mm to 11.6 mm, and the diameter of the dissolution zone of dissolved inorganic phosphorus measured on day 3 reached 12 mm to 13.9 mm.

[0088] Use a hole puncher to make three holes in the inorganic phosphorus culture medium, and inoculate 30 μL of the bacterial solution of Acinetobacter severii DE-5 isolated and purified in Example 1. Set up three parallel experiments, labeled as sample 7, sample 8 and sample 9, and culture at 28°C for 1 to 3 days. The diameter of the dissolution zone is measured at 1 day and 3 days respectively. Figure 13 As shown, A is the effect of dissolving cellulose and dissolving phosphorus circle for 1 day, and B is the effect of dissolving cellulose and dissolving phosphorus circle for 3 days. The measurement results of the diameter of the dissolving cellulose circle are as follows Figure 16 As shown, the diameter of the dissolution zone of the dissolved cellulose measured on day 1 reached 7.35 mm to 10.19 mm, and the diameter of the dissolution zone of the dissolved cellulose measured on day 3 reached 7.69 mm to 10.89 mm.

[0089] These results indicate that Acinetobacter severii DE-5 has the ability to dissolve organic phosphorus, inorganic phosphorus and cellulose, and the effect of dissolving inorganic phosphorus is more obvious.

[0090] Quantitative determination of soluble organic phosphorus and inorganic phosphorus: Add the corresponding volume of phosphorus standard solution to a 50mL volumetric flask, dilute with water to about 30mL, add 5mL of molybdenum antimony anticolorimetric agent, adjust the volume, wait for 30min, read the value, and draw a working curve. Use Zhang Xiangsheng's determination method to determine the phosphorus content. Molybdenum antimony anticolorimetric method is used to measure the effective phosphorus content in the supernatant. Add the corresponding volume of standard phosphorus solution to a 50mL volumetric flask, add 2 drops of dinitrophenol as an indicator, adjust the pH value with dilute sulfuric acid and 10% NaOH solution, add 5ml of molybdenum antimony anticolorimetric agent, adjust the volume to the scale, and make the standard phosphorus concentrations 0.2, 0.4, 0.6, 0.8, and 1.0mg / L respectively. After reacting for 30min, use UV-5500 ultraviolet-visible spectrophotometer to compare the color at 720nm, and draw a standard curve based on the results, such as Figure 17 shown.

[0091] The strain isolated and purified in Example 1 was activated. The bacterial suspension was inoculated into an organophosphorus liquid medium at a 1% inoculum rate and cultured at 30°C for 1 and 3 days at a shaker speed of 150 r / min. 6 ml of the organophosphorus medium solution was digested with 1 ml of potassium persulfate, and 3 ml was added to a 50 ml volumetric flask. The colorimetric method was the same as above. Three shake flasks were set up for the same strain and the assay was repeated three times. The phosphorus content of the bacterial suspension measured after 1 and 3 days in liquid organophosphorus medium is shown in Table 3.

[0092] The strain isolated and purified in Example 1 was activated. The bacterial suspension was inoculated into an inorganic phosphorus liquid medium at a 1% inoculum rate. The culture was shaken at 150 r / min and incubated at 30°C for 1 and 3 days. After digesting 6 ml of the inorganic phosphorus medium solution with 1 ml of potassium persulfate, 3 ml was added to a 50 ml volumetric flask. The colorimetric method was the same as above. Three shake flasks were set up for the same strain, and the results were averaged. The phosphorus content of the bacterial suspension measured after 1 and 3 days in the liquid inorganic phosphorus medium is shown in Table 4.

[0093] Table 3 Organic phosphorus content of bacterial solution

[0094]

[0095] Table 4 Inorganic phosphorus content in bacterial solution

[0096]

[0097] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0098] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A Severe Acinetobacter severi ( Acinetobacter seifertii ) DE-5, characterized in that, It was deposited in the China Center for Type Culture Collection on June 19, 2024, with the deposit number CCTCC NO: M20241302.

2. A bacterial agent, characterized in that The active ingredient is the Acinetobacter severii DE-5 described in claim 1.

3. The use of the bacterial agent in the degradation of organic phosphorus according to claim 2, characterized in that: The organophosphorus is at least one of ethoprophos and lecithin.

4. The use of the microbial agent in degrading inorganic phosphorus according to claim 2, characterized in that: The inorganic phosphorus is tricalcium phosphate.

5. Use of the bacterial agent as claimed in claim 2 in degrading cellulose.

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