A strain of Bacillus saffron and its application in petroleum hydrocarbon degradation

Abstract

The invention relates to a Bacillus safensis CY1, which is deposited in the General Microbiology Center of China Microbiological Culture Collection Committee, with the address of Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 27876. The strain has high efficiency in degrading diesel. Under the condition of 30 DEG C, for diesel content of less than 1% in an evaluation medium, the diesel degradation rate in 72 hours reaches more than 99%. The strain is moderately salt-tolerant, and the number of live bacteria is almost not affected when growing under salinity conditions of less than 5%.

Description

Technical Field

[0001] The invention relates to a strain of Bacillus saffron and application thereof in the field of wastewater and soil treatment and restoration, belonging to the technical field of environmental microorganisms. Background Art

[0002] Petroleum hydrocarbons are one of the main components of petroleum, mainly composed of alkanes and aromatic hydrocarbons, and also contain a small amount of sulfur-containing and nitrogen-containing compounds. Improper management or sudden leakage accidents during the exploration, mining, smelting, transportation, use and storage of petroleum will inevitably cause petroleum hydrocarbon pollution in the surrounding environment. Alkanes in petroleum hydrocarbons gradually deposit into the soil, groundwater and oceans, which can easily cause persistent environmental pollution. Polycyclic aromatic hydrocarbons in aromatic hydrocarbons are difficult to degrade and are carcinogenic. They will enter the human body through the food chain and threaten human life and health.

[0003] The sources of oily wastewater are very wide. In addition to the large amount of oily wastewater discharged by the petroleum extraction and processing industry, solid fuel thermal processing, wool washing wastewater in the textile industry, tanning wastewater in light industry, railway and transportation industry, slaughtering and food processing industry, and emulsions produced by turning process in the machinery industry will all discharge oily wastewater.

[0004] At present, the treatment methods for petroleum hydrocarbon pollution are mainly divided into physical methods, chemical methods and biological methods. Among them, biological treatment technology has many advantages such as low secondary pollution, high removal efficiency, thorough removal of pollutants, and high economic performance. Therefore, it has become a research hotspot in modern wastewater treatment technology and has received more and more attention.

[0005] At present, scientists have screened out a variety of strains with petroleum hydrocarbon degradation functions, such as Acinetobacter sp., Pseudomonas sp., Corynebacterium sp., Micrococcus sp., Arthrobacter sp., Alcaligenes sp. and other common petroleum hydrocarbon degradation bacteria. However, the degradation effect of these strains on petroleum hydrocarbons is still not outstanding enough, so it is necessary to screen or construct strains with more outstanding petroleum hydrocarbon degradation effects. Summary of the invention

[0006] In view of the current situation that the existing petroleum hydrocarbon degradation strains are not effective enough, the present invention provides a Bacillus saffron and its application in the field of wastewater and soil treatment and restoration. The strain can utilize petroleum hydrocarbons as a carbon source to degrade petroleum hydrocarbons in water bodies and soil.

[0007] A strain of Bacillus safensis CY1 is deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration, at the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with a deposit number of CGMCC No. 27876 and a deposit date of July 13, 2023. Its 16s rDNA is shown in SEQ ID NO: 1. Unless otherwise specified, the Bacillus safensis described in the present invention refers to the Bacillus safensis CY1 strain.

[0008] The beneficial effects of the bacillus saffron provided by the present invention are:

[0009] 1) The strain has high efficiency in degrading diesel. At 30°C, the diesel degradation rate in the evaluation medium with a diesel content of less than 1% can reach more than 99% in 72 hours;

[0010] 2) The strain has a high ability to degrade petroleum hydrocarbons. For batik wastewater with an initial oil content of 110 mg / L, the petroleum hydrocarbon degradation rate was 96% in five days, and for petrochemical sludge contaminated soil with an initial oil content of 2%, the petroleum hydrocarbon degradation rate was 98% in five weeks;

[0011] 3) The strain is moderately salt-tolerant. When growing under salinity conditions below 5%, the number of viable bacteria is almost unaffected, but it cannot tolerate salinity above 6%;

[0012] 4) The strain has a simple culture method, fast growth rate and strong environmental adaptability, high safety, does not damage the original environment, and has no secondary pollution.

[0013] The petroleum hydrocarbons described in the present invention include but are not limited to normal alkanes, branched alkanes, cycloalkanes and aromatic hydrocarbons.

[0014] The present invention also claims a microbial agent comprising the above-mentioned Bacillus saffron.

[0015] Preferably, the fermentation method of Bacillus sabdariffa comprises the following steps:

[0016] (1) Primary seed culture: Under sterile conditions, Bacillus sabdariffa was inoculated into an enrichment medium, and cultured at 25-35° C. and 150-300 rpm for 12-48 h to obtain a primary seed culture solution of Bacillus sabdariffa;

[0017] (2) Secondary seed culture: Under sterile conditions, the primary seed culture solution of Bacillus sabdariffa is inoculated into an enrichment medium at an inoculum amount of 1-10 vol%, and cultured at 25-35° C. and 150-300 rpm for 12-48 h to obtain a secondary seed culture solution of Bacillus sabdariffa;

[0018] (3) Fermentation: After the fermentation medium in the fermentation tank is disinfected, the secondary seed culture solution of Bacillus sabdariffa obtained in step (2) is inoculated into the fermentation medium at an inoculum amount of 5-10 vol%, the temperature is controlled at 25-35° C., the rotation speed is 150-300 rpm, and the fermentation is carried out under the condition of a ventilation ratio of 1:(1-2). When the dissolved oxygen begins to rise, the fermentation is stopped to obtain a fermentation solution of Bacillus sabdariffa.

[0019] The ventilation ratio described in the present invention refers to the ratio of the volume of air introduced into the fermentation tank per minute to the total volume of the fermentation liquid.

[0020] The enrichment medium is composed of: 5-15 g / L peptone, 3-8 g / L yeast extract or yeast powder, 5-15 g / L sodium chloride, the solvent is water, and the pH is 6-8.

[0021] Preferably, the composition of the enrichment medium is: 10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride, the solvent is water, and the pH is 7-7.5.

[0022] Furthermore, the fermentation medium is composed of: carbon source 10-30 g / L, nitrogen source 5-20 g / L, PO4 3- 0.8-1.5g / L, K + 0.5-1.0g / L, Mg 2+ 0.05-0.5g / L, Na + 0.1-0.5g / L, Mn 2+ 0.03-0.2g / L, Ca 2+ 0.01-0.05g / L, solvent is water, pH=6~8.

[0023] Preferably, the PO4 3- The source is one or more of dipotassium hydrogen phosphate, potassium dihydrogen phosphate, and diammonium hydrogen phosphate, wherein K + The source is one or more of dipotassium hydrogen phosphate, potassium dihydrogen phosphate, potassium sulfate, potassium chloride, and potassium nitrate, wherein the Mg 2+ The source is one or more of magnesium sulfate and magnesium chloride, wherein the Na + The source is one or more of sodium chloride, sodium nitrate or sodium sulfate, and the Mn 2+ The source is one or more of manganese sulfate monohydrate, manganese sulfate tetrahydrate, manganese nitrate or manganese chloride, wherein the Ca 2+ The source is a combination of one or more of calcium chloride and calcium nitrate.

[0024] Furthermore, the carbon source is selected from one or more of glucose, sucrose, starch, sodium acetate or sodium citrate.

[0025] Furthermore, the nitrogen source is selected from one or more of yeast extract powder, peptone, corn steep liquor powder, ammonium sulfate, diammonium hydrogen phosphate or potassium nitrate.

[0026] In actual application, the form of the final Bacillus subtilis product can be determined according to actual use and storage needs. When a liquid product is needed, the fermentation broth can be diluted to the required concentration and used directly. When a solid product is needed, the fermentation broth can be centrifuged to obtain bacterial sludge, and then spray drying or freeze drying can be used to obtain solid bacterial powder.

[0027] The present invention also claims a method for purifying sewage or soil using the activated solution of Bacillus sabogensis or a microbial agent, comprising the step of applying the activated solution of Bacillus sabogensis or a microbial agent containing Bacillus sabogensis to sewage or soil.

[0028] Furthermore, the concentration of petroleum hydrocarbons in the wastewater is below 1%, preferably below 0.5%.

[0029] Furthermore, the salinity of the wastewater is below 5%, preferably below 3%, and most preferably below 2%.

[0030] Furthermore, the inoculation amount of the activated solution of Bacillus fusus or the microbial agent is above 1000 ppm, preferably 1000-20000 ppm, and most preferably 1000-10000 ppm.

[0031] Furthermore, the concentration of petroleum hydrocarbons in the soil is below 10%, preferably below 5%, and most preferably below 2%.

[0032] The present invention also claims to protect the application of the above-mentioned Bacillus fusiformis and microbial agent in the field of sewage or soil treatment and restoration.

[0033] Preferably, the Bacillus sabdariffa and the microbial agent are used to degrade petroleum hydrocarbons in sewage or wastewater or soil; more preferably, the Bacillus sabdariffa and the microbial agent are used to degrade diesel in sewage or wastewater or soil. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is a colony photo of Bacillus saffron;

[0035] Figure 2 This is a photo of the Bacillus subtilis colony magnified 100 times under a microscope. DETAILED DESCRIPTION

[0036] The principles and features of the present invention are described below in conjunction with examples. The examples are only used to explain the present invention and are not used to limit the scope of the present invention.

[0037] The compositions of the various culture media used in the examples are as follows:

[0038] Enrichment medium: 1g potassium hydrogen phosphate, 1g potassium dihydrogen phosphate, 1g ammonium sulfate, 0.2g magnesium sulfate, 1g potassium nitrate, 0.01g calcium chloride, 1mL trace element solution, dilute to 1L, pH = 7;

[0039] Basic culture medium: peptone 10g, yeast powder 5g, sodium chloride 10g, water 1000mL, pH = 7;

[0040] Evaluation culture medium: 1g potassium hydrogen phosphate, 1g potassium dihydrogen phosphate, 1g ammonium sulfate, 0.2g magnesium sulfate, 1g potassium nitrate, 0.01g calcium chloride, 1mL trace element solution, dilute to 1L, pH = 7, adjust different diesel concentrations as needed;

[0041] Trace element solution: 1.5 g ferric chloride, 0.1 g manganese sulfate, 70 mg zinc chloride, 2 mg copper chloride, 30 mg nickel chloride, 200 mg cobalt chloride, 5 mg sodium molybdate, make up to 1 L, pH = 7.

[0042] Example 1. Screening, separation and purification of Bacillus sabdariffa

[0043] 1. Water sample enrichment

[0044] 100mL of water sample was taken from the aerobic pool of a chemical plant in Shandong, 10mL was inoculated in a conical flask containing 100mL of enrichment medium, 0.05vol% diesel was added to the enrichment medium, and the culture was shaken at 30℃ and 200r / min for 5 days. The diesel content was measured and the pH was adjusted every other day. When the diesel content dropped below 50% of the original concentration, it was transferred to the next level of enrichment, that is, 10mL of the culture solution after culture was taken and inoculated again in 100mL of enrichment medium. The concentration of diesel was increased by 0.05vol% each time, and the culture was repeated 4 times until the diesel concentration in the enrichment medium was 0.2vol%.

[0045] 2. Screening and Isolation of Strain

[0046] Take 0.5 mL of the last culture medium and dilute it with sterile water to 10 -4 , 10 -5 , 10 -6 , 10 -7 and 10 -8 times, spread the diluted culture solution on a nutrient solid culture medium plate, and then culture it in a 30℃ incubator. After the colonies grow to an appropriate size, observe the morphological characteristics of the colonies, pick single colonies for streak purification, and after three generations of purification, store them on a 4℃ slant. A total of 6 strains were isolated and named CY1~CY5.

[0047] 3. Rescreening

[0048] The five strains obtained in the initial screening were inoculated into the basal culture medium, shaken and cultured at 30°C for 24 h, 10 mL of the culture solution was taken and centrifuged at 5000 rpm for 10 min, the supernatant was discarded, and then inoculated into 100 mL of the evaluation culture medium (diesel content of 0.2 vol%), shaken and cultured at 30°C. Each experimental group was set up in triplicate, and sterile water was used as a blank control. The changes in the diesel content in the culture medium were regularly detected. The results are shown in Table 1.

[0049] Table 1 Evaluation results of the strains obtained from the initial screening on diesel

[0050]

[0051]

[0052] From the data in Table 1, it can be seen that strain CY1 has the strongest removal effect on diesel, and its ability to degrade diesel is significantly better than that of the other four strains.

[0053] Example 2. Identification of bacterial species

[0054] 1. Morphological observation

[0055] The strain CY1 was inoculated on a nutrient solid medium plate and the morphological characteristics of the colonies were observed. The colony photos of the CY1 strain are shown in Figure 1 , 100 times microscope magnification photo see Figure 2 , its colonies are dot-shaped, opaque, milky white, with neat edges, clear and smooth, and a dry surface.

[0056] 2. Molecular Biological Identification

[0057] The genomic DNA of strain CY1 was extracted and used as a template to amplify 16S rDNA.

[0058] 16S rDNA Primers:

[0059] 16S rDNA-F:5'-TATGTTGCCCTTGTCGTGG-3',

[0060] 16S rDNA-R:5'-CCGCGTTTTGTAGGCCCTCGT-3'.

[0061] The PCR product was extracted and DNA sequencing was performed using a sequencer ABI3730-XL. The spliced ​​sequence file was compared with the data in the NCBI 16S database using the NCBI Blast program to obtain the species information with the greatest sequence similarity to the species to be tested. It was found that it belonged to the genus Bacillus and was identified as Bacillus safensis, which was named Bacillus safensis CY1.

[0062] Example 3. Salinity tolerance test of Bacillus sabdariffa

[0063] Under sterile conditions, Bacillus CY1 was inoculated into a 250 mL conical flask containing 100 mL of basal culture medium, and the strain was activated by shaking and culturing in a shaker at 30°C and 200 rpm for 24 hours to obtain an activated liquid of Bacillus CY1. After testing, the number of viable bacteria in the activated liquid was 8.3 billion CFU / mL.

[0064] Using the basal medium (salinity of 1%) as the basis, after adding additional NaCl 0g, 1g, 2g, 3g, 4g, 5g, 6g, 7g, 8g, 9g to the Erlenmeyer flask containing 100mL of the basal medium, the salinity gradient was set to 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% and 10%. Take 5mL of the activation solution of Bacillus sabio respectively, inoculate it into the culture medium with different salinity gradients, and then place it in a shaking incubator at 30℃ for shaking culture, and measure its OD regularly. 600 The values ​​and viable counts were used to determine the growth extent of the strain. The results are shown in Table 2.

[0065] Table 2 OD of Bacillus saffron cultured under different salinity conditions 600 Value and viable count

[0066]

[0067] From the data in Table 2, it can be seen that strain CY1 can grow normally at salinity 1-5%, 1% is its optimal growth salinity, and the growth condition of the strain when the salinity is lower than 5% is significantly better than when the salinity is higher than 5%.

[0068] Example 4. Degradation efficiency of Bacillus sabio under different diesel content

[0069] Under sterile conditions, Bacillus CY1 was inoculated into a 250 mL conical flask containing 100 mL of basal culture medium, and the strain was activated by shaking and culturing in a shaker at 30°C and 200 rpm for 24 hours to obtain an activated solution of Bacillus CY1. After testing, the number of viable bacteria in the activated solution was 8.7 billion CFU / mL.

[0070] Evaluation culture media with diesel volume contents of 0.5%, 1%, 2%, 3%, 4% and 5% were prepared respectively, and the activated solution of Bacillus shafornien was inoculated into 100 mL of each sterilized evaluation culture medium at an inoculation rate of 1 vol%, and then placed in a shaker at 30°C and 200 rpm for shaking culture. Three parallels were set up in each group, and sterile water was added to the evaluation culture medium as a blank control. The experimental grouping arrangement is shown in Table 3, and the diesel content in each culture medium was detected every 24 hours. The results are shown in Table 4.

[0071] Table 3 Diesel content in the evaluation medium of each experimental group

[0072]

[0073] Table 4 Changes in diesel content in the evaluation medium of each experimental group and control group

[0074]

[0075]

[0076] From the data in Table 4, it can be seen that when the diesel content is below 1%, Bacillus CY1 shows excellent degradation effect, and the 72h diesel degradation rate reaches more than 99%. However, when the diesel content is above 1%, the maximum 72h degradation rate is only 53.1%, and as the diesel content increases, the degradation efficiency of CY1 gradually decreases. Therefore, this result shows that Bacillus CY1 can tolerate 1% diesel, but has poor tolerance to diesel above 1%. Example 5. Fermentation process of Bacillus CY1

[0077] The fermentation method of Bacillus fusus comprises the following steps:

[0078] 1) Pick the slant seeds of Bacillus sabio with an inoculation loop, inoculate into 100 mL of basal culture medium, and culture at 25-35° C. and 150-300 rpm for 18 h to obtain a primary seed solution;

[0079] 2) inoculating the cultured primary seed solution into 1 L of basal culture medium at an inoculum amount of 1-10 vol%, and culturing at 25-35° C. and 150-300 rpm for 18 h to obtain a secondary seed solution;

[0080] 3) Under sterile conditions, the secondary seed liquid is inoculated into a fermentation medium at an inoculum amount of 5-10 vol%, wherein the fermentation medium formula is: glucose 10 g / L, sucrose 10 g / L, yeast extract 5 g / L, ammonium sulfate 1.5 g / L, diammonium hydrogen phosphate 0.5 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, sodium chloride 0.5 g / L, manganese sulfate 0.2 g / L, calcium chloride 0.05 g / L, and the temperature is controlled at 25-35° C., the rotation speed is 150-300 rpm, and the ventilation ratio is 1: (1-2). When the dissolved oxygen begins to rise, the fermentation is stopped to obtain a fermentation liquid of Bacillus sabdariffa. After testing, the viable count of the fermentation liquid is 19.4 billion CFU / mL.

[0081] Example 6. Treatment effect of Bacillus saffron on oil-containing petrochemical wastewater

[0082] Bacillus CY1 was inoculated into a 250 mL conical flask containing 100 mL of basal culture medium under sterile conditions, and cultured in a shaker at 30°C and 200 rpm for 24 h to activate the strain, thereby obtaining an activated solution of Bacillus CY1. After testing, the number of viable bacteria in the activated solution was 8.6 billion CFU / mL.

[0083] The oily wastewater of a petrochemical in Dongying, Shandong, had an initial oil content of 2890 mg / L and a pH of 7.8 before treatment. 100 mL of this oily wastewater was placed in two identical 250 mL conical flasks. The experimental group was inoculated with the strain activation solution according to 1% of the volume of the oily wastewater, and the control group was inoculated with the same volume of sterile water. The control conditions were 30 ° C, 200 rpm shaking in a shaker, and the pH value was natural. During the reaction process, samples were taken every 48 hours after 24 hours, and the results are shown in Table 5.

[0084] Among them, the oil detection method is implemented in accordance with "GB / T16488-1996 Water Quality Determination of Petroleum and Animal and Vegetable Oils-Infrared Photometry".

[0085] Table 5 Changes of oil content in petrochemical oily wastewater over time

[0086]

[0087] From the data in Table 5, it can be seen that within six days after the addition of the strain activation solution, the oil degradation rate was 54%, which indicates that the Bacillus sabdariffa of the present invention has a certain ability to degrade high-concentration petroleum hydrocarbons.

[0088] Example 7. Treatment effect of Bacillus saffron on oil-containing batik wastewater

[0089] Bacillus CY1 was inoculated into a 250 mL conical flask containing 100 mL of basal culture medium under sterile conditions, and the strain was activated by shaking culture in a shaker at 30°C and 200 rpm for 24 h to obtain an activation solution. After testing, the number of viable bacteria in the activation solution was 8.6 billion CFU / mL.

[0090] The oil-containing batik wastewater in Shandong Province has an initial oil content of 110 mg / L and a pH of 7.8 before treatment. 100 mL of this oil-containing wastewater was placed in two identical 250 mL conical flasks. The experimental group was inoculated with the strain activation solution according to 1% of the wastewater volume, and the control group was inoculated with the same volume of sterile water. The control conditions were 30°C, 200 rpm shaking in a shaker, and natural pH. Samples were taken every 24 hours during the reaction, and the results are shown in Table 6.

[0091] Among them, the oil detection method is implemented in accordance with "GB / T16488-1996 Water Quality Determination of Petroleum and Animal and Vegetable Oils-Infrared Photometry".

[0092] Table 6 Changes of oil content in oily batik wastewater over time

[0093]

[0094] From the data in Table 6, it can be seen that within five days after the addition of the strain activation solution, the degradation rate of petroleum hydrocarbons was 96%, which indicates that the Bacillus sabdariffa of the present invention has a high efficiency in the degradation of petroleum hydrocarbons.

[0095] Example 8. Treatment effect of Bacillus saffron on oil-bearing soil

[0096] Bacillus CY1 was inoculated into a 250 mL conical flask containing 100 mL of basal culture medium under sterile conditions, and the strain was activated by shaking culture in a shaker at 30°C and 200 rpm for 24 h to obtain an activated solution of Bacillus CY1. After testing, the number of viable bacteria in the activated solution was 8.8 billion CFU / mL.

[0097] The soil in a petrochemical plant in Yantai was contaminated by oil sludge. The initial oil content before treatment was 2%, the detection value was 20200 mg / L, and the pH was 8.2.

[0098] 100g of the oil-bearing soil was placed in two identical 250mL beakers. The experimental group was inoculated with the strain activation solution according to 10% of the mass of the oil-bearing soil, and the control group was inoculated with the same mass of sterile water. The control condition was treated in a 30°C constant temperature incubator. The pH value was adjusted between 7 and 8 before the bacteria were added, and the pH was natural in the later stage. Water was added at any time during the reaction process, and nitrogen and phosphorus elements were added to ensure that the CNP ratio was 100:10:1. Samples were taken every 7 days, and the results are shown in Table 7.

[0099] Table 7 Changes of oil content in oil-bearing soil over time

[0100]

[0101] From the data in Table 7, it can be seen that the degradation rate of petroleum hydrocarbons is 97.9% five weeks after the addition of the strain activation solution, which indicates that the Bacillus sabdariffa provided by the present invention has a high efficiency in the degradation of petroleum hydrocarbons.

[0102] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A strain of Bacillus safensis, characterized in that: It is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number: CGMCC No.27876.

2. A microbial agent, characterized in that: The active ingredient comprises the Bacillus sabdariffa according to claim 1.

3. The fermentation method of Bacillus sabdariffa according to claim 1, characterized in that: The steps include: (1) Primary seed culture: Under sterile conditions, Bacillus sabdariffa was inoculated into an enrichment medium, and cultured at 25-35° C. and 150-300 rpm for 12-48 h to obtain a primary seed culture solution of Bacillus sabdariffa; (2) Secondary seed culture: Under sterile conditions, the primary seed culture solution of Bacillus sabdariffa is inoculated into an enrichment medium at an inoculum amount of 1-10 vol%, and cultured at 25-35° C. and 150-300 rpm for 12-48 h to obtain a secondary seed culture solution of Bacillus sabdariffa; (3) Fermentation: After the fermentation medium in the fermentation tank is disinfected, the secondary seed culture solution of Bacillus sabdariffa obtained in step (2) is inoculated into the fermentation medium at an inoculum amount of 5-10 vol%, the temperature is controlled at 25-35° C., the rotation speed is 150-300 rpm, and the fermentation is carried out under the condition of a ventilation ratio of 1:(1-2). When the dissolved oxygen begins to rise, the fermentation is stopped to obtain a fermentation solution of Bacillus sabdariffa.

4. The fermentation method according to claim 3, characterized in that The composition of the enrichment medium is: 5-15 g / L peptone, 3-8 g / L yeast extract or yeast powder, 5-15 g / L sodium chloride, the solvent is water, pH=6-8; The fermentation medium is composed of: 10-30 g / L carbon source, 5-20 g / L nitrogen source, PO4 3- 0.8-1.5g / L, K + 0.5-1.0g / L, Mg 2+ 0.05-0.5g / L, Na + 0.1-0.5g / L, Mn 2+ 0.03-0.2g / L, Ca 2+ 0.01-0.05g / L, solvent is water, pH=6-8.

5. The fermentation method according to claim 4, characterized in that The carbon source is selected from one or more of glucose, sucrose, starch, sodium acetate or sodium citrate; The nitrogen source is selected from one or more of yeast extract powder, peptone, corn steep liquor powder, ammonium sulfate, diammonium hydrogen phosphate or potassium nitrate.

6. A method for purifying wastewater or soil, characterized in that: The method comprises the steps of applying the activation solution of Bacillus sabogensis according to claim 1 or the microbial agent according to claim 2 to sewage or soil, wherein the Bacillus sabogensis and the microbial agent are used to degrade petroleum hydrocarbons in sewage or soil.

7. The method according to claim 6, characterized in that The concentration of petroleum hydrocarbons in the wastewater is less than 1%.

8. The method according to claim 7, characterized in that The concentration of petroleum hydrocarbons in the wastewater is below 0.5%.

9. The method according to claim 6, characterized in that The concentration of petroleum hydrocarbons in the soil is below 10%.

10. The method according to claim 9, characterized in that The concentration of petroleum hydrocarbons in the soil is below 5%.

11. The method according to claim 10, characterized in that The concentration of petroleum hydrocarbons in the soil is below 2%.

12. The method according to any one of claims 6 to 8, characterized in that: The salinity of the wastewater is below 5%.

13. The method according to claim 12, characterized in that The salinity of the wastewater is below 3%.

14. The method according to claim 13, characterized in that The salinity of the wastewater is below 2%.

15. Use of the Bacillus sabogensis of claim 1 and the microbial agent of claim 2 in the field of sewage or soil treatment and restoration, wherein the Bacillus sabogensis and the microbial agent are used to degrade petroleum hydrocarbons in sewage or soil.

16. The use according to claim 15, characterized in that The bacillus saffron and the microbial agent are used for degrading diesel in sewage or soil.

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