Method for promoting bacillus soil microorganisms to actively absorb oxygen to degrade long-chain alkane

Abstract

The invention discloses a method for promoting bacillus soil microorganisms to actively absorb oxygen to degrade long-chain alkanes, which comprises the following steps: preparing a catalyst: uniformly mixing oil-containing soil, a carbon source, citric acid, ferric salt, hydrogen peroxide and distilled water, filtering, concentrating clear liquid to obtain a concentrated crystal substance, centrifugally cleaning the crystal substance, and drying to obtain the catalyst; the method comprises the following steps: mixing a catalyst with oil-containing soil, hydrogen peroxide and distilled water, and oscillating on a shaking table for Fenton pre-oxidation; after pre-oxidation, intermittent oscillation oxygenation is carried out, bioremediation is carried out on the petroleum-contaminated soil at normal temperature, and the three petroleum bacteria in the soil are activated. Through pre-oxidation treatment, consumption of soluble organic carbon by microorganisms in soil is stimulated, the active oxygen uptake process of the microorganisms is accelerated, and the carbon metabolism process is accelerated. Finally, the bacillus-like microorganisms capable of degrading the petroleum are activated, so that the aim of degrading the petroleum hydrocarbon in the soil for a long time is fulfilled.

Description

Technical Field

[0001] The invention belongs to the field of biological restoration of petroleum-contaminated soil and relates to a method for promoting Bacillus soil microorganisms to actively absorb oxygen and degrade long-chain alkanes. Background Art

[0002] With the rapid development of the global economy, the consumption of oil and its related products has increased significantly. However, accidental leakage into the environment during oil extraction, transportation, storage and production is inevitable. Oil leakage into the soil will seriously degrade the physical structure and chemical composition of the soil, and even pose a serious threat to human health. Therefore, the treatment of oil-contaminated soil is urgent. At present, bioremediation technology is widely used due to its environmental protection and low cost. However, the simple bioremediation cycle is long, and the removal efficiency of petroleum hydrocarbons is low.

[0003] At present, the main methods of bioremediation of petroleum-contaminated soil are: 1) Biostimulation combined with bioremediation. Some scholars have used sodium α-olefin sulfonate combined with Pseudomonas to strengthen the remediation of petroleum-contaminated soil. The remediation period is 100 days, and the long-chain alkane remediation efficiency is as high as 53%. In the later stage of bioremediation, the degradation of long-chain alkanes gradually decreases from 80 days to 100 days. 2) Pre-oxidation combined with bioremediation. Some scholars used hydrogen peroxide of different concentrations for Fenton pre-oxidation, and then carried out 80 days of bioremediation on soil containing a large amount of crude oil. The TPH removal rate can reach up to 64.4%. However, most of the current reports on the use of bioremediation of petroleum-contaminated soil mainly focus on the removal of petroleum pollutants in the early stage of bioremediation. However, there is little research on how to make bioremediation technology efficiently remove petroleum pollutants in the later stage of bioremediation and achieve long-term remediation of petroleum-contaminated soil.

[0004] In addition, studies in recent years have shown that enzymes secreted by microorganisms play a vital role in the process of degrading organic pollutants. The dissolved organic carbon content in the soil is a key factor affecting the secretion of enzymes by microorganisms. Some scholars have found that by adding rapeseed to the soil, the dissolved organic carbon content in the soil is increased, which significantly promotes the process of microbial enzyme secretion. In addition, it was found that enzyme activity is proportional to the consumption of dissolved organic carbon by microorganisms. It can be seen that the large-scale consumption of dissolved organic carbon can promote the production of microbial enzymes, thereby promoting the active oxygen absorption process of microorganisms and accelerating the carbon metabolism rate.

[0005] Therefore, how to design a method to accelerate the carbon metabolism rate by promoting the active oxygen absorption process of microorganisms so as to degrade petroleum hydrocarbons in the soil for a longer period of time has become an urgent problem to be solved. Summary of the invention

[0006] In order to solve the above-mentioned defects existing in the prior art, the purpose of the present invention is to provide a method for promoting the active oxygen absorption of Bacillus soil microorganisms to degrade long-chain alkanes. By performing appropriate pre-oxidation treatment on petroleum-contaminated soil, the consumption of dissolved organic carbon by microorganisms in the soil can be stimulated, the active oxygen absorption process of the microorganisms is accelerated, and thus the carbon metabolism process is accelerated. Finally, the Bacillus microorganisms that can degrade petroleum are activated, thereby achieving the purpose of long-term degradation of petroleum hydrocarbons in the soil.

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

[0008] In one aspect of the present invention, a method for promoting active oxygen uptake by Bacillus soil microorganisms to degrade long-chain alkanes is provided, comprising the following steps:

[0009] The oil-bearing soil, carbon source, citric acid, iron salt, hydrogen peroxide and distilled water are mixed uniformly according to a mass ratio of (5-10): (10-20): (0.2-0.5): (0.1-0.2): (3-6): (5-10), filtered, and the clear liquid is concentrated to obtain concentrated crystals, the crystals are washed by centrifugation, and dried to obtain a catalyst;

[0010] The catalyst is mixed with oil-containing soil, hydrogen peroxide and distilled water in a mass ratio of (1-2): (5-10): (3-6): (30-50), and the mixture is shaken on a shaker for Fenton pre-oxidation; after pre-oxidation, the mixture is shaken intermittently for oxygenation, and the petroleum-contaminated soil is bioremediated at room temperature.

[0011] Preferably, the carbon source is chitosan, glucose or fructose.

[0012] Preferably, the iron salt is ferrous sulfate or ferric sulfate.

[0013] Preferably, the petroleum hydrocarbon content in the oil-bearing soil is 11.5-12.5 g / kg.

[0014] Preferably, the Fenton pre-oxidation is carried out by shaking on a shaker at a rotation speed of 150 to 180 rpm / min for 10 to 12 hours.

[0015] Preferably, the mixture is intermittently oscillated on an oscillator with a rotation speed of 150 to 180 r / min, and oxygenated once every 6 to 8 hours.

[0016] Preferably, the bioremediation of petroleum contaminated soil is carried out at room temperature for 80 to 100 days.

[0017] Preferably, the three oil-degrading bacteria activated in the soil include Acinetobacter, Paenibacillus and Microbacterium.

[0018] Another aspect of the present invention provides a catalyst used in the method, comprising the following raw materials in a mass ratio:

[0019] Oil-containing soil 5-10; carbon source 10-20; citric acid 0.2-0.5; iron salt 0.1-0.2; hydrogen peroxide 3-6; distilled water 5-10.

[0020] The present invention adopts the above technical solution, which has the following beneficial effects:

[0021] The present invention adds an appropriate amount of catalyst to perform Fenton pre-oxidation to stimulate the consumption of dissolved organic carbon by microorganisms in the soil, accelerate the active oxygen absorption process of the microorganisms, and thus accelerate the carbon metabolism process. Finally, the Bacteroidetes microorganisms that can degrade petroleum are activated, thereby achieving the purpose of long-term degradation of petroleum hydrocarbons in the soil. Furthermore, the present invention measures the concentration of total petroleum hydrocarbons and each alkane, the concentration of dissolved organic carbon, the concentration of three nitrogens (ammonia nitrogen, nitrate nitrogen, nitrite nitrogen), the types of microorganisms, and the number of total bacteria and petroleum bacteria in the soil.

[0022] After pre-oxidation combined with bioremediation, it was found that the removal of long-chain alkanes reached 8846 mg / kg; in addition, three types of petroleum-degrading bacteria were activated, including Acinetobacter, Paenibacillus, and Microbacterium. The proportion of petroleum bacteria can reach up to 94.24%. During the bioremediation process, dissolved organic carbon was consumed in large quantities, and the microbial activity was not less than 0.15 mgCO2 / kg soil. Thus, the long-term degradation of soil long-chain alkanes was achieved. This method can significantly improve the removal efficiency of petroleum hydrocarbons, shorten the time of bioremediation, and save energy, and has great engineering application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The drawings described herein are used to provide a further understanding of the present invention, constitute a part of the present application, and do not constitute an improper limitation of the present invention. In the drawings:

[0024] Figure 1a This is a graph showing the degradation of total petroleum hydrocarbons and long-chain alkanes in soil by oxidation-biodegradation combined with biodegradation in Example 1 and Comparative Example 1;

[0025] Figure 1b This is a graph showing the degradation of total petroleum hydrocarbons and long-chain alkanes in soil by oxidation-combined biodegradation in Example 2 and Comparative Example 2;

[0026] Figure 1c This is a graph showing the degradation of total petroleum hydrocarbons and long-chain alkanes in soil by oxidation-combined biodegradation in Example 3 and Comparative Example 3;

[0027] Figure 2aThis is a graph showing the consumption of dissolved organic carbon during the biodegradation process of Example 1 and Comparative Example 1;

[0028] Figure 2b This is a graph showing the consumption of dissolved organic carbon during the biodegradation process of Example 2 and Comparative Example 2;

[0029] Figure 2c It is a graph of the consumption of dissolved organic carbon during the biodegradation process of Example 3 and Comparative Example 3;

[0030] Figure 3a This is a graph showing the change in the proportion of petroleum bacteria during the biodegradation process of Example 1 and Comparative Example 1;

[0031] Figure 3b This is a graph showing the change in the proportion of petroleum bacteria during the biodegradation process of Example 2 and Comparative Example 2;

[0032] Figure 3c This is a graph showing the change in the proportion of petroleum bacteria during the biodegradation process of Example 3 and Comparative Example 3;

[0033] Figure 4 This is an analysis chart of the changes in microbial CO2 production during biodegradation;

[0034] Figure 5 This is a graph showing the changes in abundance of microbial species in soil after bioremediation. DETAILED DESCRIPTION

[0035] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments. The exemplary embodiments and descriptions of the present invention are used to explain the present invention but are not intended to limit the present invention.

[0036] A method for promoting Bacillus soil microorganisms to actively absorb oxygen and degrade long-chain alkanes provided by an embodiment of the present invention comprises the following steps:

[0037] 1) Preparation of catalyst:

[0038] Add oil-bearing soil, carbon source (chitosan, glucose or fructose), citric acid, iron salt (ferrous sulfate or ferric sulfate), 30% hydrogen peroxide and distilled water in a mass ratio of (5-10): (10-20): (0.2-0.5): (0.1-0.2): (3-6): (5-10) into a 250 mL conical flask, mix well, filter the solid in the conical flask, concentrate the clear liquid to obtain concentrated crystals, wash the crystals with distilled water by centrifugation for 3-5 times, and finally dry them in a muffle furnace to obtain a catalyst;

[0039] 2) The catalyst prepared in step 1) is added to a 250 mL conical flask with oil-containing soil, 30% hydrogen peroxide and distilled water in a mass ratio of (1-2): (5-10): (3-6): (30-50) and mixed evenly. The conical flask is then placed on a shaking table with a rotation speed of 150-180 rpm / min and shaken for 10-12 hours to perform a Fenton pre-oxidation experiment; after pre-oxidation, the conical flask is placed on a shaking table and shaken and oxygenated once every 6-8 hours on an oscillator with a rotation speed of 150-180 r / min, and the bioremediation of petroleum-contaminated soil is performed at room temperature for 80-100 days.

[0040] Among them, the content of petroleum hydrocarbons in petroleum-contaminated soil is 11.5-12.5g / kg.

[0041] The following determination method is used to verify the removal performance of this method for petroleum hydrocarbons.

[0042] 20 mL of dichloromethane was added to each soil sample, and the sample was shaken for 12 h, then ultrasonicated for 15 minutes and mechanically vibrated for 30 minutes (250 rpm). The oil solution was extracted three times in a row using a separatory funnel and then dehydrated by anhydrous sodium sulfate (prebaked in an oven at 105 ° C for 2 hours). The final volume of the extracted oil solution was adjusted to 50 mL for TPH extraction and analysis. Total organic carbon and total inorganic carbon were determined using Shimadzu TOC (TOC-L, Shimadzu, Kyoto, Japan), where soluble organic carbon was equal to total organic carbon minus total inorganic carbon. The supernatant in the reaction bottle was filtered through a 0.45 μm filter membrane. Ammonia nitrogen, nitrate nitrogen and nitrite nitrogen were analyzed by ultraviolet spectrophotometry. Soil microbial counts were determined using the plate dilution method, and the growth rate of the microorganisms was obtained by taking the logarithm to characterize the bacterial respiratory activity by CO2 production.

[0043] The effect of the method of the present invention in promoting the active oxygen uptake by microorganisms to degrade petroleum hydrocarbons in a long term can be seen from the following aspects.

[0044] (1) Promoting the active oxygen uptake of microorganisms to degrade petroleum hydrocarbons

[0045] After pre-oxidation combined with bioremediation, the total petroleum hydrocarbon removal in the soil was as high as 8839 mg / kg, which was 1.3 times that of the group without activated microorganisms and active oxygen absorption. In addition, the petroleum bacteria in the active oxygen absorption group had a better degradation effect on long-chain alkanes, with a removal amount of 4512 mg / kg.

[0046] (2) Analysis of dissolved organic carbon consumption by microorganisms during biodegradation

[0047] The consumption of dissolved organic carbon and ammonia nitrogen in the active oxygen uptake group at each stage of biodegradation was significantly higher than that in the non-active oxygen uptake group; the consumption of dissolved organic carbon in each cycle of the active oxygen uptake group was 4.04, 4.98, 1.62, 2.67, and 2.34 times that of the non-active oxygen uptake group, respectively. In the active oxygen uptake group and the active oxygen uptake group, the microbial activity coefficient was linearly related to the consumption of dissolved organic carbon. As the consumption of dissolved organic carbon increased, the microbial activity coefficient also increased. This may be due to the higher consumption of dissolved organic carbon in the active oxygen uptake group, which promoted the active oxygen uptake process of microorganisms and increased microbial activity. It can be seen that the high consumption of dissolved organic carbon by microorganisms in the active oxygen uptake group promoted the synthesis of enzymes, promoted the active oxygen uptake process of microorganisms, accelerated the carbon metabolism process of microorganisms, and thus increased the degradation rate of long-chain alkanes.

[0048] (3) Analysis of the degradation ability of petroleum bacteria

[0049] The average degradation of long-chain alkanes per unit of petroleum bacteria in the active oxygen inhalation group was 43.58 (mg / (kg·log CFU / g)), while the average degradation of long-chain alkanes per unit of petroleum bacteria in the non-active oxygen inhalation group was 28.40 (mg / (kg·log CFU / g)). In addition, the highest percentage of logarithmic petroleum bacteria in the active oxygen inhalation group was 94.24%, while the highest percentage in the active oxygen inhalation group was only 83.34%.

[0050] (4) Analysis of changes in microbial communities during bioremediation

[0051] Biodiversity analysis showed that the dominant bacterial genera in the soil of the active oxygen uptake group were mainly Acinetobacter, Paenibacillus, and Microbacterium, with a relative abundance of 8.02%, 10.26%, and 16.64%, respectively. The dominant bacterial genera in the soil of the non-active oxygen uptake group were mainly Rothia, Streptococcus, and Neisseria; this is the main reason why the active oxygen uptake group can degrade long-chain alkanes in the soil for a long time.

[0052] The present invention is further illustrated by means of specific examples below.

[0053] Example 1

[0054] 1) Preparation of catalyst:

[0055] Add oil-bearing soil with a petroleum hydrocarbon content of 11.5 g / kg, chitosan, citric acid, ferrous sulfate, 30% hydrogen peroxide and distilled water in a mass ratio of 5:15:0.4:0.1:5:10 into a conical flask. After mixing evenly, filter the solid in the conical flask, concentrate the clear liquid to obtain concentrated crystals, wash the crystals with distilled water by centrifugation for 3 times, and finally dry them in a muffle furnace to obtain the catalyst;

[0056] 2) Add the catalyst prepared in step 1) into a conical flask with oil-containing soil, 30% hydrogen peroxide and distilled water in a mass ratio of 1:7:6:40 and mix well. Then place the conical flask on a shaker at a speed of 150rpm / min and shake for 12 hours to perform a Fenton pre-oxidation experiment. After pre-oxidation, place the conical flask on a shaking shaker and shake and oxygenate it once every 8 hours on an oscillator at a speed of 150r / min. The bioremediation process of petroleum-contaminated soil is carried out at room temperature for 100 days. The changes in the abundance of microbial species in the soil after bioremediation are shown in Figure 5 shown.

[0057] After the reaction, the consumption of dissolved organic carbon in the biodegradation process reached 16723 mg / kg; the petroleum bacteria accounted for the highest proportion of 94.23%; the microbial activity reached a maximum of 0.27 mgCO2 / kg soil; and the long-chain alkanes (C 25 ~C 30 ) removal amount is 3886mg / kg; that is, this method promotes the active oxygen absorption and rapid reproduction of petroleum bacteria, and can efficiently degrade petroleum hydrocarbons in the soil. Figure 1a , 2a , 3a, Figure 4 shown.

[0058] Example 2

[0059] 1) Preparation of catalyst:

[0060] Add oil-bearing soil with a petroleum hydrocarbon content of 12 g / kg, glucose, citric acid, ferric sulfate, 30% hydrogen peroxide and distilled water in a mass ratio of 7:20:0.2:0.2:6:8 into a conical flask. After mixing evenly, filter the solid in the conical flask, concentrate the clear liquid to obtain concentrated crystals, wash the crystals with distilled water by centrifugation for 4 times, and finally dry them in a muffle furnace to obtain the catalyst;

[0061] 2) The catalyst prepared in step 1) was added to a conical flask with oil-containing soil, 30% hydrogen peroxide and distilled water in a mass ratio of 2:5:4:50 and mixed evenly. The conical flask was then placed on a shaker at a speed of 160 rpm / min and shaken for 11 hours to perform a Fenton pre-oxidation experiment. After pre-oxidation, the conical flask was placed on a shaking shaker for intermittent shaking and oxygenation, and the bioremediation process of petroleum-contaminated soil was carried out at room temperature for 80 days. The changes in the abundance of microbial species in the soil after bioremediation are shown in Figure 5 shown.

[0062] After the reaction, the consumption of dissolved organic carbon in the biodegradation process reached 19419 mg / kg; petroleum bacteria accounted for the highest proportion of 93.88%; the long-chain alkanes (C 25 ~C 30 ) removal amount is 3939mg / kg; that is, this method promotes the active oxygen absorption and rapid reproduction of petroleum bacteria, and can efficiently degrade petroleum hydrocarbons in the soil. Figure 1b , 2b , as shown in 3b.

[0063] Example 3

[0064] 1) Preparation of catalyst:

[0065] Add oil-bearing soil with a petroleum hydrocarbon content of 12.5 g / kg, fructose, citric acid, ferrous sulfate, 30% hydrogen peroxide and distilled water in a mass ratio of 10:10:0.5:0.15:3:5 into a conical flask. After mixing evenly, filter the solid in the conical flask, concentrate the clear liquid to obtain concentrated crystals, wash the crystals with distilled water by centrifugation for 5 times, and finally dry them in a muffle furnace to obtain the catalyst;

[0066] 2) The catalyst prepared in step 1) was added to a 250 mL conical flask with oil-containing soil, 30% hydrogen peroxide and distilled water in a mass ratio of 2:10:3:30 and mixed evenly. The conical flask was then placed on a shaker at a speed of 180 rpm / min and shaken for 6 hours to perform a Fenton pre-oxidation experiment. After pre-oxidation, the conical flask was placed on a shaking shaker for intermittent shaking and oxygenation, and the bioremediation process of petroleum-contaminated soil was carried out at room temperature for 90 days. The changes in the abundance of microbial species in the soil after bioremediation are shown in Figure 5 shown.

[0067] After the reaction, the consumption of dissolved organic carbon in the biodegradation process reached 16274 mg / kg; the petroleum bacteria accounted for the highest proportion of 93.65%; the long-chain alkanes (C 25 ~C 30 ) removal amount is 4512mg / kg; that is, this method promotes the active oxygen absorption and rapid reproduction of petroleum bacteria, and can efficiently degrade petroleum hydrocarbons in the soil. Figure 1c ,2c , as shown in 3c.

[0068] Comparative Example 1

[0069] Oil-containing soil, 30% hydrogen peroxide and distilled water were added to a 250 mL conical flask at a mass ratio of 5:3:30 and mixed evenly. The conical flask was then placed on a shaker at a speed of 180 rpm / min for 12 hours for Fenton pre-oxidation experiment. After pre-oxidation, the conical flask was placed on a shaking shaker for intermittent shaking and oxygenation, and the bioremediation process of petroleum-contaminated soil was carried out at room temperature for 100 days.

[0070] After the reaction, the consumption of dissolved organic carbon in the biodegradation process was only 5641 mg / kg; the proportion of petroleum bacteria was very low, only 83.32% at most; the highest microbial activity was only 0.18 mgCO2 / kg soil; and the long-chain alkanes were only degraded by 2048 mg / kg in the end, without promoting the active oxygen absorption process of microorganisms, resulting in low efficiency of soil long-chain alkane degradation. Figure 1a , 2a , 3a, Figure 4 shown.

[0071] Comparative Example 2

[0072] Oil-containing soil, 30% hydrogen peroxide and distilled water were added to a 250 mL conical flask in a mass ratio of 6:5:45 and mixed evenly. The conical flask was then placed on a shaker at a speed of 150 rpm / min for 10 hours for Fenton pre-oxidation experiment. After pre-oxidation, the conical flask was placed on a shaking shaker for intermittent shaking and oxygenation, and the bioremediation process of petroleum contaminated soil was carried out at room temperature for 100 days.

[0073] After the reaction, the consumption of dissolved organic carbon in the biodegradation process was only 4656 mg / kg; the proportion of petroleum bacteria was very low, with the highest being only 80.96%; and finally, only 1886 mg / kg of long-chain alkanes were degraded, without promoting the active oxygen absorption process of microorganisms, resulting in low efficiency of soil long-chain alkane degradation. Figure 1b , 2b , as shown in 3b.

[0074] Comparative Example 3

[0075] Oil-containing soil, 30% hydrogen peroxide and distilled water were added to a 250 mL conical flask in a mass ratio of 10:6:45 and mixed evenly. The conical flask was then placed on a shaker at a speed of 150 rpm / min for 10 hours for Fenton pre-oxidation experiment. After pre-oxidation, the conical flask was placed on a shaking shaker for intermittent shaking and oxygenation, and the bioremediation process of petroleum contaminated soil was carried out at room temperature for 100 days.

[0076] After the reaction, the consumption of dissolved organic carbon in the biodegradation process was only 5535 mg / kg; the proportion of petroleum bacteria was very low, with the highest being only 82.32%; and finally, only 2040 mg / kg of long-chain alkanes were degraded, without promoting the active oxygen absorption process of microorganisms, resulting in low efficiency of soil long-chain alkane degradation. Figure 1c , 2c , as shown in 3c.

[0077] By comparing Examples 1 to 3 with Comparative Examples 1 to 3, it can be seen that the consumption of dissolved organic carbon in the biodegradation process of the active oxygen absorption group is not less than 16274 mg / kg; the highest proportion of petroleum bacteria is not less than 93.65%; and the degradation of long-chain alkanes is not less than 3886 mg / kg. It can be seen that after promoting active oxygen absorption of microorganisms by this method, the consumption of dissolved organic carbon, the proportion of petroleum bacteria, the activity of microorganisms and the degradation of long-chain alkanes in the soil have been greatly improved. Therefore, the method of the present invention has good application potential and is worthy of promotion and application.

[0078] It should be noted that the user information (including but not limited to user device information, user personal information, etc.) and data (including but not limited to data used for analysis, stored data, displayed data, etc.) involved in this application are all information and data authorized by the user or fully authorized by all parties, and the collection, use and processing of relevant data must comply with relevant laws, regulations and standards of relevant countries and regions.

[0079] The present invention is not limited to the above-mentioned embodiments. On the basis of the technical solution disclosed in the present invention, technicians in this field can make some substitutions and deformations to some technical features therein according to the disclosed technical content without creative labor, and these substitutions and deformations are all within the protection scope of the present invention.

Claims

1. A method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms, characterized in that: The following steps are involved: The oil-bearing soil, carbon source, citric acid, iron salt, hydrogen peroxide and distilled water are mixed uniformly according to a mass ratio of (5-10): (10-20): (0.2-0.5): (0.1-0.2): (3-6): (5-10), filtered, and the clear liquid is concentrated to obtain concentrated crystals, the crystals are washed by centrifugation, and dried to obtain a catalyst; The catalyst is mixed with oil-containing soil, hydrogen peroxide and distilled water in a mass ratio of (1-2): (5-10): (3-6): (30-50), and the mixture is shaken on a shaker for Fenton pre-oxidation; after pre-oxidation, the mixture is shaken intermittently for oxygenation, and the petroleum-contaminated soil is bioremediated at room temperature.

2. The method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms according to claim 1, characterized in that: The organic carbon source is chitosan, glucose or fructose.

3. The method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms according to claim 1, characterized in that: The iron salt is ferrous sulfate or ferric sulfate.

4. The method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms according to claim 1, characterized in that: The petroleum hydrocarbon content in the oil-bearing soil is 11.5-12.5 g / kg.

5. The method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms according to claim 1, characterized in that: Fenton pre-oxidation was performed by shaking on a shaker at a speed of 150-180 rpm / min for 10-12 hours.

6. The method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms according to claim 1, characterized in that: Oscillate intermittently on an oscillator with a rotation speed of 150-180 r / min and oxygenate once every 6-8 hours.

7. The method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms according to claim 1, characterized in that: Bioremediation of petroleum contaminated soil takes 80 to 100 days at room temperature.

8. The method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms according to claim 1, characterized in that: The three oil-degrading bacteria activated in the soil include Acinetobacter, Paenibacillus and Microbacterium.

9. The method for promoting active oxygen absorption and degradation of long-chain alkanes by Bacillus soil microorganisms according to claim 1, characterized in that: The highest consumption of dissolved organic carbon in the process of biodegradation in the soil was 19419 mg / kg; the highest proportion of petroleum bacteria reached 94.24%; the bacterial activity was not less than 0.15 mgCO2 / kg soil; the degradation amount of long-chain alkanes was 4512 mg / kg.

10. A catalyst used in the method according to any one of claims 1 to 9, characterized in that: The raw materials include the following mass ratios: Oil-containing soil 5-10; carbon source 10-20; citric acid 0.2-0.5; iron salt 0.1-0.2; hydrogen peroxide 3-6; distilled water 5-10.

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