Salt-tolerant petroleum hydrocarbon degrading bacterium and application thereof
By using bacteria agents prepared by Acinetobacter junii H1, medium and long-chain alkanes in petroleum are efficiently degraded in high-salt environments, the problem of difficult degradation in the prior art is solved, and effective repair of oil pollution is achieved.
Patent Information
- Application Number
- CN202311271396.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-09-28
- Publication Date
- 2025-03-28
AI Technical Summary
The prior art is difficult to efficiently degrade medium and long chain alkanes in petroleum, especially in high salt environments, and there are fewer bacteria agents used for practical applications in existing alkane degradation strains.
A bacteria agent can be prepared by using Acinetobacter junii H1 as the active ingredient, which can efficiently degrade petroleum, alkanes and waxes in a high-salt environment.
Acinetobacter junii H1 can quickly grow and efficiently degrade C10-C32 alkanes, especially in high salt environments, and is suitable for oil pollution repair in water and soil.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of environmental pollutant microbial treatment, and relates to a salt-tolerant petroleum hydrocarbon degrading bacterium and application thereof. Background Art
[0002] After the Industrial Revolution, the continuous development of the oil industry has brought benefits to society, but it has also caused a series of environmental pollution problems. Oil is widely used as an energy source and is a very important basic energy source. However, the oil refining process, oil waste residue treatment and oil leakage have had a serious impact on the environment, especially the petrochemical industry and refineries have produced a large amount of solid sludge waste, which has caused soil pollution in the surrounding area.
[0003] Petroleum contains a lot of components, such as water, wax, asphaltene, fat, colloid, metal, solid, etc. In addition, the highest content is 40-52% of alkanes and 28%-31% of aromatic hydrocarbons. The medium- and long-chain alkanes in alkanes have poor water solubility and are difficult to degrade, so they are more harmful to the environment. Once they enter the soil, they will form waxy substances that adhere to the roots of plants, causing the roots of plants to rot; they can also enter the human body through breathing, contact, enrichment in the food chain, etc., thus endangering human health.
[0004] At present, there are many remediation methods for the medium- and long-chain alkane pollution problem, such as physical, chemical, and biological remediation. The microbial remediation method of bioremediation is also considered to be one of the very important means of controlling petroleum hydrocarbon pollution at this stage. Microorganisms can use medium- and long-chain alkanes for growth and metabolism, and convert them into harmless carbon dioxide and release them into the air, thereby reducing the content of petroleum hydrocarbons in the environment. In addition, most of the existing contaminated sites have high salt concentrations, so there is a great need for a microorganism that can tolerate high salt concentrations and can efficiently degrade medium- and long-chain alkanes. There are many alkane-degrading bacteria that have been reported, but there are few strains that can be prepared into bacterial agents for the degradation of medium- and long-chain petroleum hydrocarbons for practical application. Summary of the invention
[0005] The technical problem to be solved by the present invention is how to efficiently and quickly degrade petroleum, such as alkanes, waxes, etc.
[0006] In order to solve the above technical problems, the present invention provides an Acinetobacter junii H1, whose deposit number in the General Microbiological Center of China National Microbiological Culture Collection is CGMCC No.27582.
[0007] The present invention also provides a bacterial agent, the active ingredient of the bacterial agent is the Acinetobacter junii H1.
[0008] The above-mentioned microbial agent has any of the following uses:
[0009] 1) Degradation of petroleum;
[0010] 2) Degradation of alkanes;
[0011] 3) Degradation of wax;
[0012] 4) Remediation of petroleum, alkane or wax pollution in water or soil.
[0013] In the above-mentioned microbial agent, the inorganic phosphorus may be at least one of calcium hydrogen phosphate, phosphate rock, iron phosphate, aluminum phosphate and calcium phosphate; and the organic phosphorus may be lecithin.
[0014] In the above-mentioned bacterial agent, the bacterial agent may further include a carrier. The carrier may be a solid carrier or a liquid carrier. The solid carrier may be a mineral material, a plant material or a polymer compound; the mineral material may be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica and diatomaceous earth; the plant material may be at least one of corn flour, soybean flour and starch; the polymer compound may be polyvinyl alcohol and / or polyglycol. The liquid carrier may be an organic solvent, vegetable oil, mineral oil or water; the organic solvent may be decane and / or dodecane. In the bacterial agent, the active ingredient may be present in the form of cultured living cells, fermentation broth of living cells, filtrate of cell culture or a mixture of cells and filtrate. The dosage form of the composition may be a variety of dosage forms, such as liquid, emulsion, suspension, powder, granule, wettable powder or water-dispersible granule.
[0015] As required, a surfactant (such as Tween 20, Tween 80, etc.), a binder, a stabilizer (such as an antioxidant), a pH adjuster, etc. may also be added to the bacterial agent.
[0016] The present invention also provides any of the following uses of the Acinetobacter junii H1 or the bacterial agent:
[0017] 1) Degradation of petroleum;
[0018] 2) Degradation of alkanes;
[0019] 3) Degradation of wax;
[0020] 4) Remediation of petroleum, alkane or wax pollution in water or soil;
[0021] 5) Preparation of degraded petroleum products;
[0022] 6) preparing degradation alkane products;
[0023] 7) preparing degraded wax products;
[0024] 8) Preparation of products for repairing petroleum, alkane or wax pollution in water or soil.
[0025] The petroleum, alkane and wax can be petroleum, alkane and wax in water or soil respectively.
[0026] The degradation can be carried out in an environment containing 0.1-30 g / L NaCl.
[0027] In the above application, the alkane may be a C10-C32 alkane or a C16-C32 alkane; and the wax may be paraffin.
[0028] The present invention also provides a method for degrading petroleum or its components, the method comprising: utilizing the Acinetobacter junii H1 to treat petroleum or its components, or to treat a sample containing petroleum or its components, so as to achieve degradation of the petroleum or its components.
[0029] In the above method, the components of the petroleum may be alkanes or waxes.
[0030] In the above method, the alkane may be a C10-C32 alkane or a C16-C32 alkane; and the wax is paraffin.
[0031] In the above method, the petroleum, alkane and wax can be petroleum, alkane and wax in water or soil, respectively.
[0032] In the above method, the degradation can be carried out in an environment containing 0.1-30 g / L NaCl.
[0033] The present invention also provides a method for culturing the Acinetobacter junii H1, which comprises the step of culturing the Acinetobacter junii H1 in a culture medium for culturing Acinetobacter junii.
[0034] The present invention also provides a method for preparing the bacterial agent, comprising the following steps: using the Acinetobacter junii H1 as an active ingredient to obtain the bacterial agent.
[0035] Acinetobacter junii H1 of the present invention can utilize the alkane (such as hexadecane, octadecane, eicosane, tetracosane, octacosane, dotriacontane) of C10-C32 and the solid paraffin containing more than hexadecane as carbon source growth, and can also be degraded. When hexadecane is the sole carbon source alone, the bacterial strain can grow fast, and the first day just can reach the stable period, and the degradation rate in 1-4 days is the fastest; When hexacosane is the sole carbon source, the stable period can be reached on the 6th day. In addition, Acinetobacter junii H1 can degrade multiple alkanes simultaneously in water bodies, preferentially utilizes low-chain alkanes, and starts to utilize long-chain alkanes after reaching a certain cell density, and the above-mentioned bacterial strain can utilize long-chain alkanes alone, and can also utilize medium-long chain alkanes simultaneously. Acinetobacter junii can degrade C16, C18, C20, C24, C28, and C32 alkanes by 87.4%, 97.8%, 79.92%, 77.65%, and 71.9% respectively within 7 days; and can tolerate medium and high concentrations of monoalkanes and mixed alkanes, such as degrading 3.85g / L of n-hexadecane to 1.74g / L within 12 days, and degrading 1.8g / L of n-octacosane to 0.45g / L within 9 days, and degrading 6.77g / L of mixed hydrocarbons in the soil to 2.92g / L, with a degradation rate of 43.11%. The strain can also degrade solid paraffin (a mixture of solid alkanes above C20) by about 15% at a NaCl concentration of 30g / L, and can degrade by about 40% at medium and low concentrations. In water and soil experiments, Acinetobacter junii also has a high degradation ability for alkanes. The Acinetobacter junii of the present invention has a good application prospect in the pollution remediation of water and soil petroleum and its components.
[0036] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.
[0037] Description of biological material deposit
[0038] Abbreviation of depository unit: CGMCC
[0039] Name of depository: China National Microbiological Culture Collection Administration General Microbiology Center
[0040] Address of the depository: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Postal Code: 100101
[0041] Date of deposit: June 8, 2023
[0042] CGMCC Registration Number: CGMCC No.27582
[0043] Classification name: Acinetobacter junii
[0044] Strain number: H1 BRIEF DESCRIPTION OF THE DRAWINGS
[0045] Figure 1 This is the phylogenetic tree of Acinetobacterjunii H1.
[0046] Figure 2 Schematic diagram of the degradation rate of different alkanes by Acinetobacter juniiH1.
[0047] Figure 3 Schematic diagram of the degradation rate of solid paraffin by Acinetobacter juniiH1 at different salt concentrations.
[0048] Figure 4 Schematic diagram of the ability of Acinetobacter juniiH1 to degrade n-hexadecane.
[0049] Figure 5 Schematic diagram of the ability of Acinetobacter juniiH1 to degrade n-octacosane.
[0050] Figure 6 Schematic diagram of the degradation of mixed alkanes by Acinetobacter juniiH1. DETAILED DESCRIPTION
[0051] The experimental methods in the following examples are conventional methods unless otherwise specified, and are performed according to the techniques or conditions described in the literature in the art or according to the product instructions. The materials, reagents, instruments, etc. used in the following examples are all commercially available unless otherwise specified. The quantitative tests in the following examples are all repeated at least three times, and the results are averaged.
[0052] The composition of basal mineral salt medium (MSM) B is as follows: the solvent is water, and the solutes and their concentrations in the medium are K2HPO4·3H2O 2.1 g / L, KH2PO4 0.4 g / L, NaCl 5 g / L, (NH4)2SO4 0.5 g / L, 0.5 g / LNaNO3, CaCl2 0.1 g / L, MgCl2 0.1 g / L, 1 mL / L 1000× trace element stock solution, and 50 μL / L 20000× vitamin stock solution.
[0053] The composition of the basic inorganic salt medium (MSM) C is as follows: K2HPO4·3H2O 2.1g / L, KH2PO4 0.4g / L, NaCl5g / L, (NH4)2SO42g / L, CaCl2 0.1g / L, MgCl2 0.1g / L, 1mL / L1000× trace element stock solution, 50μL / L20000× vitamin stock solution.
[0054] 1000× trace element mother solution: the solvent is water. Each liter of 1000× trace element mother solution contains 5g EDTA, 2g FeSO4·7H2O, 0.1g ZnSO4·7H2O, 0.03g MnCl2·4H2O, 0.3g H3BO3, 0.2g CoCl2·6H2O, 0.01g CuCl2·2H2O, 0.02g NiCl2·6H2O, and 0.03g Na2MoO4·H2O.
[0055] 20000× vitamin mother solution: The solvent is water. Each liter of 20000× vitamin mother solution contains 2 mg biotin (Biotin), 2 mg folic acid (Folic), 5 mg thiamine hydrochloride VB1 (ThiamineHCl), and 5 mg riboflavin.
[0056] 5g / L n-octadecane mother liquor: a solution with a n-octadecane concentration of 5g / L obtained by dissolving n-octadecane in n-hexane; 5g / L n-eicosane mother liquor: a solution with a n-eicosane concentration of 5g / L obtained by dissolving n-eicosane in n-hexane; 5g / L n-tetracosane mother liquor: a solution with a n-eicosane concentration of 5g / L obtained by dissolving n-tetracosane in n-hexane; 5g / L n-octacosane mother liquor: a solution with a n-eicosane concentration of 5g / L obtained by dissolving n-octacosane in n-hexane; 10g / L n-octacosane mother liquor: a solution with a n-eicosane concentration of 10g / L obtained by dissolving n-octacosane in n-hexane; 5g / L n-dotriacontane mother liquor: a solution with a n-eicosane concentration of 5g / L obtained by dissolving n-dotriacontane in n-hexane.
[0057] Example 1, Isolation and Identification of Acinetobacter juniiH1
[0058] Separation from crude oil contaminated soil, using basic inorganic salt medium B with 2g / L crude oil as separation medium, taking 2mL of contaminated soil leachate and adding it to the separation medium, after culturing for about 15 days, transfer it to a new separation medium to enrich functional strains, and after continuing to culture for 15 days, take the enrichment solution and dilute it with sterile water for 10 -3 , 10 -4 , 10 -5 , 10 -6 The strains were plated on R2A medium at equal multiples and cultured at 30°C for 3-5 days. Single colonies of different shapes, colors and sizes were picked out for pure culture. The pure cultured strains were added in sequence to the basic inorganic salt medium B with n-hexadecane as the only carbon source. It was found that strain H1 grew in n-hexadecane. Subsequently, it was added to the basic inorganic salt medium B with petroleum as the only carbon source, and it was found that it could also grow in petroleum and produce obvious turbidity.
[0059] The 16S rRNA of strain H1 was amplified and sequenced, and the results showed that its sequence was SEQ ID No. 1 in the sequence list. Based on the sequence, the phylogenetic tree of the strain was constructed using MEGA11.0 ( Figure 1 ). The results showed that it clustered with Acinetobacter junii ATCC 17908, and strain H1 was identified as Acinetobacter junii and named Acinetobacter junii H1.
[0060] Acinetobacter junii H1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration (CGMCC, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, 100101) on June 8, 2023, with the deposit number CGMCC No. 27582. It is hereinafter referred to as A.junii H1.
[0061] Example 2: Single-bacteria fermentation and determination of alkane degradation range
[0062] (1) Liquid culture medium A was prepared, sterilized and cooled, and then inoculated with A. junii H1 prepared in Example 1. The seed solution was cultured at a constant temperature of 30° C. and a rotation speed of 160 rpm.
[0063] The formula of liquid culture medium A is as follows: yeast extract 0.5 g / L, peptone 0.5 g / L, casamino acid 0.5 g / L, glucose 0.5 g / L, K2HPO4·3H2O 0.39 g / L, MgSO4·7H2O 0.05 g / L, sodium pyruvate 0.3 g / L, soluble starch 0.5 g / L, NaCl 5 g / L, distilled water to 1 L, pH about 7.0.
[0064] (2) The seed solution obtained in step (1) was centrifuged at 4500 rpm, and the bacterial cells were collected; the collected bacterial cells were washed twice with basic inorganic salt medium B, and then resuspended with basic inorganic salt medium B and inoculated into basic inorganic salt medium B with different length alkanes (n-hexadecane, n-eicosane, n-tetracosane, n-octacosane, n-dotriacontane) as the sole carbon source. The OD of A. junii H1 in the inoculated system was 600 The concentration of 0.08 was used and the culture was carried out at 30°C and 160 rpm for 7 days.
[0065] The basic inorganic salt medium B with alkanes of different lengths as the sole carbon source are n-hexadecane inorganic salt medium B, n-eicosane inorganic salt medium B, n-tetracosane inorganic salt medium B, n-octacosane inorganic salt medium B and n-dotriacontane inorganic salt medium B respectively. Among them, the n-hexadecane inorganic salt medium B is a medium in which n-hexadecane is added to the basic inorganic salt medium B, and the obtained n-hexadecane volume percentage content is 1% (about 7.7 g / L), the n-eicosane inorganic salt medium B is a medium in which 200 μL of 5g / L n-eicosane mother solution is added to the basic inorganic salt medium B, and the obtained n-eicosane content is 200 mg / L, the n-tetracosane inorganic salt medium B is a medium in which 100 μL of 5g / L n-tetracosane mother solution is added to the basic inorganic salt medium B, and the obtained n-tetracosane content is 100 mg / L, the n-octacosane inorganic salt medium B is a medium in which 100 μL of 5g / L n-octacosane mother solution is added to the basic inorganic salt medium B, and the obtained n-octacosane content is 100 mg / L, and the n-dotriacontane inorganic salt medium B is a medium in which 100 μL of 5g / L n-dotriacontane mother solution is added to the basic inorganic salt medium B, and the obtained n-dotriacontane content is 100 mg / L. When used, each culture medium is sterilized first, and then the n-hexane is evaporated.
[0066] After the cultivation is completed, n-hexadecane culture solution, n-eicosane culture solution, n-tetracosane culture solution, n-octacosane culture solution, and n-dotriacontane culture solution are obtained respectively. Then, the content of alkanes in each culture solution is detected by gas chromatography.
[0067] The n-hexadecane culture medium was extracted with n-hexane as the extractant, and anhydrous sodium sulfate was added for dehydration and then filtered to prepare the gas phase sample. The gas chromatograph used for sample detection was Agilent 7890A, the column was Agilent HP-5, the column specifications were 60m×250μm×0.25μm, the starting temperature was 70℃, and then increased to 310℃ at 10℃ / min and retained for 10min. According to this step, the n-hexadecane culture medium was replaced with n-eicosane culture medium, n-tetracosane culture medium, n-octacosane culture medium, and n-dotriacontane culture medium, and the content of alkanes in each culture medium was detected respectively.
[0068] The results showed that the strain Acinetobacter juniiH1 of Example 1 could use n-hexadecane, n-eicosane, n-tetracosane, n-octacosane and n-dotriacontane as the sole carbon source, and could degrade the above alkane components by 87.4%, 97.8%, 79.92%, 77.65%, 71.9% ( Figure 2 ).
[0069] This example shows that Acinetobacter junii H1 can be used as an alkane-degrading bacterium to utilize alkanes from C16 to C32 with a high degradation efficiency.
[0070] Example 3: Effects of different salt concentrations on the growth and alkane degradation of A. junii H1
[0071] In the experiment of testing the strain A.junii H1 of Example 1 under different salt concentrations and with solid paraffin as the sole carbon source, the OD was measured after 14 days of growth. 600 value.
[0072] Solid paraffin inorganic salt medium C: Weigh 0.2g solid paraffin and add it to the basic inorganic salt medium C, sterilize it with high pressure steam at 121℃ for 15min, and obtain a medium with a solid paraffin content of 2g / L. Adjust the concentration of NaCl in the solid paraffin inorganic salt medium C to obtain solid paraffin inorganic salt medium C with NaCl concentrations of 0.1, 5, 10, 15, 20, 25, 30 and 50g / L respectively.
[0073] The strain A.juniiH1 of Example 1 was inoculated into solid paraffin inorganic salt medium C with different NaCl concentrations. The OD of A.junii H1 in the system after inoculation was 600 is about 0.08; the obtained system was cultured at 30°C, 160rpm for 14 days.
[0074] After the culture was completed, the content of solid paraffin in each solid paraffin culture solution was detected by gas chromatography.
[0075] Solid paraffin was extracted from each solid paraffin culture solution using n-hexane as the extractant, and anhydrous sodium sulfate was added for dehydration and then filtered to prepare gas phase samples. The gas chromatograph used for sample detection was Agilent 7890A, the column was Agilent HP-5, the column size was 60m×250μm×0.25μm, the starting temperature was 70℃, then increased to 310℃ at 10℃ / min, and retained for 10min.
[0076] The results showed that the NaCl concentration of 0.1g / L-20g / L did not affect the growth of the strain using alkanes. 600 It can reach 0.7-0.8, and the NaCl concentration is slightly affected from 25g / L. 600 The growth state of the strain under 30g / L NaCl treatment was poor, and OD 600 The growth rate of the strain is linearly related to the degradation rate. The experimental results are shown in Figure 3 The degradation rate at a salt concentration of 20g / L was 36%. The strain had the highest degradation rate when treated with a NaCl concentration of 5g / L. The degradation rate gradually decreased with the increase of salt concentration. The degradation rate was 22% at 25g / L and 15% at 30g / L. The strain did not grow at 50g / L and the alkanes were not degraded at all.
[0077] This example shows that A. junii H1 can be used as a petroleum hydrocarbon degrading bacterium, can grow at a NaCl concentration of 0.1-30 g / L and can degrade a certain amount of alkanes.
[0078] Example 4: Degradation ability of monoalkanes in water
[0079] The growth of the strain A. junii H1 of Example 1 was measured with 1% of the medium-short chain alkane n-hexadecane and 2 g / L of the medium-long chain alkane n-octacosane as the sole carbon source.
[0080] The n-hexadecane inorganic salt culture medium C is a sterile culture medium with a n-hexadecane volume percentage content of 1% (about 15.4 g / L) obtained by adding n-hexadecane to the basic inorganic salt culture medium C; the n-octacosane inorganic salt culture medium C is a sterile culture medium with a n-octacosane content of 2 g / L obtained by adding 10 g / L n-octacosane mother liquor to the basic inorganic salt culture medium C.
[0081] The strain A.junii H1 was inoculated into the hexadecane inorganic salt medium C. After inoculation, the OD of A.junii H1 in the system was 600The growth rate of the strain and the amount of alkane residue were determined at 1d, 2d, 3d, 4d, 6d, 7d and 12d of culture. The method for determining the amount of alkane residue was the same as that in Example 2.
[0082] The strain A.junii H1 was inoculated into the n-octacosane inorganic salt medium C. After inoculation, the OD of A.junii H1 in the system was 600 The growth rate of the strain and the amount of alkane residue were determined at 1d, 2d, 3d, 4d, 5d, 6d, 7d, 9d and 12d of culture, and the method for determining the amount of alkane residue was the same as that in Example 2.
[0083] The experimental results are shown in Figure 4 and Figure 5 The degradation rate of n-hexadecane by A.junii H1 reached 58% at 12 days, and the degradation efficiency was fastest at 1-4 days. The number of viable bacteria in the culture medium reached 1.5×10 9 CFU / mL, the degradation rate was 32%, and then the bacterial density gradually decreased, indicating that the strain can grow well using n-hexadecane and reach a stable period within 24 hours, and then enter the decay period. On the 4th day, the number of viable bacteria dropped to 1×10 9 CFU / mL, the degradation rate reached 51%, and the degradation rate was 0.69g / d. The degradation rate of n-octacosane reached 47.12% at 12 days, and it slowly degraded from the first day and reached a plateau on the sixth day. On the sixth day, the number of viable bacteria in the culture medium could reach a maximum of 1.8×10 9 CFU / mL, and then gradually decreased, and the degradation rate could reach 36.41% on the 6th day.
[0084] This example shows that medium- and long-chain alkanes can be a good carbon source for the growth of A. junii H1, especially in n-hexadecane, where the growth rate is almost the same as that of carbon sources such as sodium acetate. However, as the carbon chain of the alkane becomes longer, the strain reaches the logarithmic growth phase more slowly.
[0085] Example 5: Degradation ability of mixed hydrocarbons in water
[0086] Mixed alkane inorganic salt medium C: A sterile culture medium obtained by adding 5g / L n-octadecane mother solution, 5g / L n-eicosane mother solution, 5g / L n-tetracosane mother solution, 5g / L n-octacosane mother solution, and 5g / L n-dotriacontane mother solution to the basic inorganic salt medium C. The total alkane content in the culture medium is 2g / L, and the contents of n-octadecane, n-eicosane, n-tetracosane, n-octacosane, and n-dotriacontane are all 0.4g / L.
[0087] The strain A.junii H1 of Example 1 was inoculated into the mixed alkane inorganic salt medium C. After inoculation, the OD of A.junii H1 in the system was 600 The growth rate of the strain and the amount of residual alkanes were determined at 1d, 2d, 3d, 4d, 5d, 7d and 13d of culture. The method for determining the amount of residual alkanes was the same as that in Example 2.
[0088] The experimental results are shown in Figure 6 In the degradation of mixed hydrocarbons, it was found that the A.junii H1 strain preferentially degraded low-chain alkanes, mainly n-octadecane and n-eicosane, which could be degraded by 79.4% and 60.7% respectively on the 13th day; n-octacosane and n-tetracosane began to degrade on the 2nd day, and could be degraded by 39.7% and 36.7% respectively on the 13th day; n-dotriacontane began to degrade on the 3rd day, and could be degraded by 42.7% on the 13th day. Therefore, this strain can efficiently degrade medium and long-chain alkanes.
[0089] This example shows that the petroleum hydrocarbon-degrading bacteria A. junii H1 can not only grow and degrade well in a single alkane, but can also grow and degrade mixed alkanes well.
[0090] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, and without the need to carry out unnecessary experimental conditions, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that the present invention can be further improved. In a word, according to the principles of the present invention, the application is intended to include any changes, uses or improvements to the present invention, including departure from the disclosed scope in the application, and changes made with conventional techniques known in the art.
Claims
1. Acinetobacter junii H1, whose deposit number in the General Microbiological Center of China Culture Collection Administration is CGMCC No.27582.
2. A bacterial agent, the active ingredient of which is the Acinetobacter junii H1 as claimed in claim 1.
3. The bacterial agent according to claim 2, characterized in that: The bacterial agent has any of the following uses: 1) Degradation of petroleum; 2) Degradation of alkanes; 3) Degradation of wax; 4) Remediation of petroleum, alkane or wax pollution in water or soil.
4. Any of the following uses of the Acinetobacter junii H1 of claim 1 or the bacterial agent of claim 2 or 3: 1) Degradation of petroleum; 2) Degradation of alkanes; 3) Degradation of wax; 4) Remediation of petroleum, alkane or wax pollution in water or soil; 5) Preparation of degraded petroleum products; 6) preparing degradation alkane products; 7) preparing degraded wax products; 8) Preparation of products for repairing petroleum, alkane or wax pollution in water or soil.
5. The use according to claim 4, characterized in that: The alkane is a C10-C32 alkane or a C16-C32 alkane; and the wax is paraffin.
6. A method for degrading petroleum or its components, comprising: The Acinetobacter junii H1 described in claim 1 is used to treat petroleum or its components, or to treat a sample containing petroleum or its components, so as to achieve degradation of petroleum or its components.
7. The method according to claim 6, characterized in that: The components of the petroleum are alkanes or waxes.
8. The method according to claim 7, characterized in that: The alkane is a C10-C32 alkane or a C16-C32 alkane; and the wax is paraffin.
9. A method for culturing the Acinetobacter junii H1 of claim 1, comprising the step of culturing the Acinetobacter junii H1 in a culture medium for culturing Acinetobacter junii.
10. A method for preparing the bacterial agent according to claim 2, comprising the following steps: using the Acinetobacter junii H1 according to claim 1 as an active ingredient to obtain the bacterial agent.