CN112795502A - Acinetobacter johnsonii WP01 and application thereof in alkane degradation - Google Patents
Acinetobacter johnsonii WP01 and application thereof in alkane degradation Download PDFInfo
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- CN112795502A CN112795502A CN202011603783.4A CN202011603783A CN112795502A CN 112795502 A CN112795502 A CN 112795502A CN 202011603783 A CN202011603783 A CN 202011603783A CN 112795502 A CN112795502 A CN 112795502A
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- acinetobacter junii
- alkane
- degradation
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/32—Hydrocarbons, e.g. oil
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Abstract
The invention relates to acinetobacter junii WP01, which is preserved in the China Center for Type Culture Collection (CCTCC) with the preservation date of 2020, 11 and 6 days, and the preservation numbers are as follows: CCTCC NO: m2020697, the preservation address is Wuhan, Wuhan university, postcode 430072. Use of acinetobacter junii WP01 for degrading alkanes. A method for degrading alkanes, comprising the step of degrading alkanes with acinetobacter junii WP 01. The acinetobacter junii WP01 has a good degradation effect on alkanes with different carbon chain lengths, wherein the degradation rate of dodecane, octadecane, docosane and dotriacontane for 120 hours exceeds 60%, and the degradation rate of dodecane is close to 80%; in the aspect of crude oil degradation, the 120-hour degradation effect of acinetobacter junii WP01 on 1g/L standard crude oil is 76%, and the data show that the strain has excellent potential for degrading petroleum pollutants in the environment.
Description
Technical Field
The invention relates to the technical field of environmental biology, in particular to Acinetobacter junii WP01 and application thereof in alkane degradation.
Background
The process of crude oil extraction and transportation, and the process of processing and using some petroleum industrial products, can cause the leakage of petroleum hydrocarbon, and further cause serious soil and water pollution problems. When petroleum pollution occurs in oceans, lakes or rivers, it causes serious water pollution, and when these water sources flow into the ground, it causes pollution of groundwater. The petroleum pollutants can form an oil film with uneven thickness on the water surface due to low density, and the oil film isolates the atmosphere and the water surface, so that the self-repairing capability of the water body is limited, and the water quality becomes black and smelly.
Physical and chemical methods are the commonly used restoration methods in petroleum pollution treatment, and mainly comprise an adsorption method, a heat treatment method, an extraction method, a washing method and a chemical oxidation method. In addition, many microorganisms can grow by using petroleum hydrocarbon as a sole carbon source and energy source, and the microorganisms are used for degrading petroleum hydrocarbon pollutants in the environment, so that the microorganisms are an ideal method.
Disclosure of Invention
The invention aims to solve the technical problem of providing Acinetobacter junii (Acinetobacter junii) WP01 and application thereof in alkane degradation so as to overcome the defects in the prior art.
The technical scheme for solving the technical problems is as follows: acinetobacter junii (Acinetobacter junii) WP01, which is preserved in the China center for type culture Collection with the preservation date of 2020, 11 and 6 days, and the preservation numbers are as follows: CCTCC NO: m2020697, the preservation address is Wuhan, Wuhan university, postcode 430072.
On the basis of the technical scheme, the invention can be further improved as follows.
Application of acinetobacter junii WP01 in alkane degradation.
A method for degrading an alkane, comprising the step of degrading an alkane with said acinetobacter junii WP 01.
Further: the method for degrading alkanes comprises the following steps:
a hydrocarbon-containing environment was inoculated with Acinetobacter johnsonii WP 01.
Further: the inoculated acinetobacter junii WP01 is a strain subjected to activation treatment.
Further: the specific steps of activating the strain are as follows:
sucking acinetobacter junii WP01, adding the acinetobacter junii WP01 into an LB culture medium, and placing the culture medium in a shaking table at the temperature of 28-30 ℃ and at the speed of 150-200 rpm for culturing for 24-30 h;
transferring the cultured acinetobacter junii WP01 into a fresh LB culture medium, and culturing for 24-30 h in a shaking table at 28-30 ℃ at 150-200 rpm.
Further: the alkane-containing environment is:
preparing alkane degradation culture media containing alkanes with different lengths, inoculating acinetobacter junii WP01, and placing the cells in a shaking table at the temperature of 28-30 ℃ and sealing and culturing at the speed of 150-200 rpm.
Further: the formula of the alkane degradation culture medium is as follows: NaH2PO42H2O 1.0g/L，(NH4)2SO4 0.5 g/L，NaNO3 0.5g/L，CaCl2 0.02g/L，MgSO4.H2O 0.2g/L，KH2PO41.0g/L, NaCl 10g/L, alkane of different lengths 0.5 g/L-1.5 g/L, and pH 7.5.
Further: the alkane-containing environment is:
dissolving original crude oil in petroleum ether with a boiling range of 30-60 ℃, removing water through anhydrous sodium sulfate treatment, placing the solution in a water bath at 60 ℃ to evaporate the petroleum ether, and then placing in a constant temperature box at 60 ℃ to remove residual petroleum ether to obtain standard oil;
preparing standard oil degradation culture media containing standard oil with different concentrations, inoculating acinetobacter junii WP01, and placing in a shaking table at 28-30 ℃ and sealing and culturing at 180-200 rpm.
Further: the formula of the standard oil degradation culture medium is as follows: NaH2PO42H2O 1.0g/L，(NH4)2SO4 0.5g/L，NaNO3 0.5g/L，CaCl2 0.02g/L，MgSO4.H2O 0.2g/L，KH2PO41.0g/L, NaCl 10g/L, standard crude oil 1 g/L-10 g/L, pH 7.5.
The invention has the beneficial effects that: the acinetobacter junii WP01 has a good degradation effect on alkanes with different carbon chain lengths, wherein the degradation rate of dodecane, octadecane, docosane and dotriacontane for 120 hours exceeds 60%, and the degradation rate of dodecane is close to 80%; in the aspect of crude oil degradation, the 120-hour degradation effect of acinetobacter junii WP01 on 1g/L standard crude oil is 76%, and the data show that the strain has excellent potential for degrading petroleum pollutants in the environment.
Drawings
FIG. 1 is a phylogenetic tree of Acinetobacter johnsonii WP 01.
Detailed Description
The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention.
As shown in FIG. 1, Acinetobacter junii (Acinetobacter junii) WP01 is preserved in China center for type culture Collection with a preservation date of 2020, 11 and 6 days and a preservation number of: CCTCC NO: m2020697, the preservation address is Wuhan, Wuhan university, postcode 430072.
Application of acinetobacter junii WP01 in alkane degradation.
A method for degrading an alkane, comprising the step of degrading an alkane with said acinetobacter junii WP 01.
The method for degrading alkane comprises the following steps:
1) activating strains:
sucking 50 mu L of acinetobacter junii WP01 bacterial liquid from a glycerol tube, adding the liquid into an LB culture medium, and placing the liquid in a shaking table at 28-30 ℃ and at 150-200 rpm for culturing for 24-30 h;
transferring the cultured acinetobacter junii WP01 into a fresh LB culture medium, and culturing for 24-30 h in a shaking table at 28-30 ℃ at 150-200 rpm;
the LB culture medium has the formula: 10g/L of peptone, 5g/L of yeast extract powder and 10g/L of sodium chloride;
2) degradation of alkanes of different lengths:
preparing alkane degradation culture media containing alkanes with different lengths, wherein the formula of the alkane degradation culture media is as follows: NaH2PO42H2O 1.0g/L，(NH4)2SO4 0.5g/L，NaNO3 0.5g/L，CaCl2 0.02g/L， MgSO4.H2O 0.2g/L，KH2PO41.0g/L, 10g/L NaCl, 1g/L of alkane with different lengths and pH of 7.5, 300mL of alkane degradation culture medium is added into a 1000mL triangular flask, acinetobacter junii WP01 is inoculated, the flask is placed in a shaking table at 30 ℃ for closed culture at 200rpm, samples are taken every 24 hours, and the alkane content in the culture medium is determined by gas chromatography, and the results are shown in the following table.
A method of degrading alkanes, comprising the steps of:
1) activating strains:
sucking 50 mu L of acinetobacter junii WP01 bacterial liquid from a glycerol tube, adding the liquid into an LB culture medium, and placing the liquid in a shaking table at 28-30 ℃ and at 150-200 rpm for culturing for 24-30 h;
transferring the cultured acinetobacter junii WP01 into a fresh LB culture medium, and culturing for 24-30 h in a shaking table at 28-30 ℃ at 150-200 rpm;
the LB culture medium has the formula: 10g/L of peptone, 5g/L of yeast extract powder and 10g/L of sodium chloride;
2) degradation of standard oil:
dissolving original crude oil in petroleum ether with a boiling range of 30-60 ℃, removing water through anhydrous sodium sulfate treatment, filtering by a sand core funnel to remove impurities, water-absorbing sodium sulfate and the like, putting the filtrate in a water bath at 60 ℃ to evaporate the petroleum ether, and then putting the filtrate in a constant temperature box at 60 ℃ to remove residual petroleum ether to obtain standard oil;
preparing standard oil degradation culture media containing standard oil with different concentrations, wherein the standard oil degradation culture media comprise the following components in parts by weight: NaH2PO42H2O 1.0g/L，(NH4)2SO4 0.5g/L，NaNO3 0.5g/L，CaCl2 0.02 g/L，MgSO4.H2O 0.2g/L，KH2PO41.0g/L, 10g/L NaCl, 1 g/L-10 g/L of standard crude oil and pH 7.5, 300mL of standard oil degradation culture medium is added into a 1000mL triangular flask, acinetobacter junii WP01 is inoculated, the flask is placed in a shaking table at 30 ℃ for closed culture at 200rpm, samples are taken every 24 hours, and the alkane content in the culture medium is measured by gas chromatography, and the results are shown in the following table.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Sequence listing
<110> Mitsu Wu design research Co., Ltd
<120> acinetobacter junii WP01 and application thereof in alkane degradation
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1385
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
gtagcgtcct ccttgcggtt agactaccta cttctggtgc acaaactccc atggtgtgac 60
gggcggtgtg tacaaggccc gggaacgtat tcaccgcggc attctgatcc gcgattacta 120
gcgattccga cttcatggag tcgagttgca gactccaatc cggactacga tcggcttttt 180
gagattagca tcacatcgct gtgtagcaac cctttgtacc gaccattgta gcacgtgtgt 240
agccctggcc gtaagggcca tgatgacttg acgtcgtccc cgccttcctc cagtttgtca 300
ctggcagtat ccttaaagtt cccatccgaa atgctggcaa gtaaggaaaa gggttgcgct 360
cgttgcggga cttaacccaa catctcacga cacgagctga cgacagccat gcagcacctg 420
tatctagatt cccgaaggca ccaatccatc tctggaaagt ttctagtatg tcaaggccag 480
gtaaggttct tcgcgttgca tcgaattaaa ccacatgctc caccgcttgt gcgggccccc 540
gtcaattcat ttgagtttta gtcttgcgac cgtactcccc aggcggtcta cttatcgcgt 600
tagctgcgcc actaaagcct caaaggcccc aacggctagt agacatcgtt tacggcatgg 660
actaccaggg tatctaatcc tgtttgctcc ccatgctttc gtacctcagc gtcagtatta 720
ggccagatgg ctgccttcgc catcggtatt cctccagatc tctacgcatt tcaccgctac 780
acctggaatt ctaccatcct ctcccatact ctagcttccc agtatcgaat gcaattccca 840
agttaagctc ggggatttca catccgactt aaaaagccgc ctacgcacgc tttacgccca 900
gtaaatccga ttaacgctcg caccctctgt attaccgcgg ctgctggcac agagttagcc 960
ggtgcttatt ctgcgagtaa cgtccactat ccaagagtat tagtctcagt agcctcctcc 1020
tcgcttaaag tgctttacaa ccataaggcc ttcttcacac acgcggcatg gctggatcag 1080
ggttcccccc attgtccaat attccccact gctgcctccc gtaggagtct gggccgtgtc 1140
tcagtcccag tgtggcggat catcctctca gacccgctac agatcgtcgc cttggtaggc 1200
ctttacccca ccaactagct aatccgactt aggctcatct attagcgcaa ggtccgaaga 1260
tcccctgctt tctcccgtag gacgtatgcg gtattagcat tcctttcgga atgttgtccc 1320
ccactaatag gcagattcct aagcattact cacccgtccg ccgctaagat aaggtgcaag 1380
cacct 1385
Claims (10)
1. The acinetobacter junii WP01 is preserved in the China center for type culture Collection with the preservation date of 2020, 11 and 6 days, and the preservation numbers are as follows: CCTCC NO: m2020697, the preservation address is Wuhan, Wuhan university, postcode 430072.
2. The use of acinetobacter junii WP01 as claimed in claim 1 for degrading alkanes.
3. A method for degrading an alkane, comprising the step of degrading an alkane with acinetobacter junii WP01 of claim 1.
4. A method according to claim 3, characterized by the steps of:
acinetobacter junii WP01 was inoculated into an environment containing alkanes.
5. The method of claim 4, wherein the inoculated acinetobacter junii WP01 is activated.
6. The method as claimed in claim 5, wherein the specific steps of activating the bacterial species are as follows:
sucking acinetobacter junii WP01, adding the acinetobacter junii WP01 into an LB culture medium, and placing the culture medium in a shaking table at the temperature of 28-30 ℃ and at the speed of 150-200 rpm for culturing for 24-30 h;
transferring the cultured acinetobacter junii WP01 into a fresh LB culture medium, and culturing for 24-30 h in a shaking table at 28-30 ℃ at 150-200 rpm.
7. A method according to any one of claims 4 to 6, wherein the alkane-containing environment is:
preparing alkane degradation culture media containing alkanes with different lengths, inoculating acinetobacter junii WP01, and placing the cells in a shaking table at the temperature of 28-30 ℃ and sealing and culturing at the speed of 150-200 rpm.
8. The method according to any one of claims 4 to 7, wherein the alkane degradation medium has a formula of: NaH2PO4 2H2O 1.0g/L，(NH4)2SO4 0.5g/L，NaNO3 0.5g/L，CaCl20.02g/L，MgSO4.H2O 0.2g/L，KH2PO41.0g/L, NaCl 10g/L, alkane of different lengths 0.5 g/L-1.5 g/L, and pH 7.5.
9. A method according to any one of claims 4 to 6, wherein the alkane-containing environment is:
dissolving original crude oil in petroleum ether with a boiling range of 30-60 ℃, removing water through anhydrous sodium sulfate treatment, placing the solution in a water bath at 60 ℃ to evaporate the petroleum ether, and then placing in a constant temperature box at 60 ℃ to remove residual petroleum ether to obtain standard oil;
preparing standard oil degradation culture media containing standard oil with different concentrations, inoculating acinetobacter junii WP01, and placing in a shaking table at 28-30 ℃ and sealing and culturing at 180-200 rpm.
10. The method of claim 9, wherein the standard oil degradation medium is formulated as: NaH2PO42H2O 1.0g/L，(NH4)2SO4 0.5g/L，NaNO3 0.5g/L，CaCl2 0.02g/L，MgSO4.H2O 0.2g/L，KH2PO41.0g/L, NaCl 10g/L, standard crude oil 1 g/L-10 g/L, pH 7.5.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114134066A (en) * | 2021-10-13 | 2022-03-04 | 西南科技大学 | High-yield lipase strain and application thereof in oil degradation |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520616A (en) * | 2016-11-07 | 2017-03-22 | 中国石油大学(北京) | Acinetobacter junii for producing bio-surfactant and application of acinetobacter junii |
| CN110846257A (en) * | 2019-12-11 | 2020-02-28 | 中冶华天工程技术有限公司 | Microbial bacterium for degrading long-chain alkane and application thereof |
| US20200239343A1 (en) * | 2019-01-30 | 2020-07-30 | Institut National De Recherche Scientifique | In situ enzymatic degradation of hydrocarbon-polluted soils |
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2020
- 2020-12-30 CN CN202011603783.4A patent/CN112795502B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520616A (en) * | 2016-11-07 | 2017-03-22 | 中国石油大学(北京) | Acinetobacter junii for producing bio-surfactant and application of acinetobacter junii |
| US20200239343A1 (en) * | 2019-01-30 | 2020-07-30 | Institut National De Recherche Scientifique | In situ enzymatic degradation of hydrocarbon-polluted soils |
| CN110846257A (en) * | 2019-12-11 | 2020-02-28 | 中冶华天工程技术有限公司 | Microbial bacterium for degrading long-chain alkane and application thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114134066A (en) * | 2021-10-13 | 2022-03-04 | 西南科技大学 | High-yield lipase strain and application thereof in oil degradation |
| CN114134066B (en) * | 2021-10-13 | 2023-04-21 | 西南科技大学 | High-yield lipase strain and application thereof in grease degradation |
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| CN112795502B (en) | 2022-08-30 |
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