CN101560483A - Lipopeptide-producing bacillus pumilus and application thereof - Google Patents
Lipopeptide-producing bacillus pumilus and application thereof Download PDFInfo
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- CN101560483A CN101560483A CNA2009100273597A CN200910027359A CN101560483A CN 101560483 A CN101560483 A CN 101560483A CN A2009100273597 A CNA2009100273597 A CN A2009100273597A CN 200910027359 A CN200910027359 A CN 200910027359A CN 101560483 A CN101560483 A CN 101560483A
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- bacillus pumilus
- lipopeptide
- bacterial strain
- cyhalothrin
- producing bacillus
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- 241000194103 Bacillus pumilus Species 0.000 title claims abstract description 45
- 108010028921 Lipopeptides Proteins 0.000 title claims abstract description 18
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- 238000006731 degradation reaction Methods 0.000 claims abstract description 27
- 241000894006 Bacteria Species 0.000 claims abstract description 24
- ZXQYGBMAQZUVMI-UNOMPAQXSA-N cyhalothrin Chemical compound CC1(C)C(\C=C(/Cl)C(F)(F)F)C1C(=O)OC(C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 ZXQYGBMAQZUVMI-UNOMPAQXSA-N 0.000 claims abstract description 23
- 239000000575 pesticide Substances 0.000 claims abstract description 9
- 239000003208 petroleum Substances 0.000 claims abstract description 8
- 230000002906 microbiologic effect Effects 0.000 claims abstract description 4
- 230000001737 promoting effect Effects 0.000 claims abstract 3
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- 239000002689 soil Substances 0.000 claims description 17
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- 229910052799 carbon Inorganic materials 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 11
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- 244000005700 microbiome Species 0.000 claims description 6
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
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Abstract
The invention discloses a lipopeptide-producing bacillus pumilus, and classification thereof is named to be Bacillus pumilus Y8A which is preserved by China Center for Type Culture Collection (CCTCC) in Wuhan University with the preservation number of CCTCCM 209108. The invention also discloses an application of the lipopeptide-producing bacillus pumilus in promoting the micro-biological degradation process of petroleum and/or cyhalothrin pesticide. Bacterial strain Y8A of the invention can produce lipopeptide biosurfactant with high efficiency and stability, thereby effectively promoting the micro-biological degradation process of petroleum and/or cyhalothrin pesticide degrading bacteria. Bacterial strain Y8A with the function of efficiently producing lipopeptide is expected to work as a new-generation bacteria and cosolvent for antibiotic production and, is widely used in agriculture, medicine, cosmetic and food industries, and can be used for restoration of pollutant environment together with other organic pollution degrading microbes. The bacterial strain is resistant to a plurality of antibiotics with easy fermentation course and convenient operation.
Description
Technical field
The invention belongs to environmental microorganism engineering field, be specifically related to a kind of lipopeptide-producing bacillus pumilus and application thereof.
Background technology
Reasons such as the exploitation of oil, transportation, storage and accidental leakage cause has up to ten million ton petroleum hydrocarbons to enter environment every year approximately, causes the severe contamination of soil, underground water, water system and ocean environment, is subjected to people's attention day by day.After petroleum pollution enters water body, the chemical oxygen demand (COD) of water body is raise, the oil film of surface layer of water has stoped airborne oxygen to spread in water again simultaneously, cause the anaerobic state in local waters, make hydrocoles dead because of anoxic, the photosynthesis of waterplant also can be destroyed, and brings serious harm to the ecosystem.Soil is infiltrated in Oil spills, equivalent surpass soil after net capacity, the oily substance of accumulation with extended residual in wherein, destroy Soil structure, influence soil permeability, the infringement plant root, hinder the breathing and the absorption of root, to the edaphophyte and the soil microorganisms ecosystem, even underground water all produces harm, has a strong impact on soil productivity and crop yield.Administer the research focus that petroleum pollution has become the countries nowadays environmentalist.
Sterilant more than 98% and 95% above weedicide all do not affact on the target organisms, but have arrived non-target organism, enter soil, among air, water and the food, destroy the ecosystem, cause the acute or chronic poisoning of people and animals and plants.The surface in contact water-soluble relatively poor, with microorganism in biodegradation process of (for example pyrethroid insecticides of synthetic such as cyhalothrin) is long-pending little because most of agricultural chemicals, make that the biological restoration process is slower, this also is a long principal element of agricultural chemicals biodegradation period.
Last decade comes, and what a class was new constantly is found and is studied by bacterium non-ribosomal synthetic lipopeptid class secondary metabolite.The bacterium lipopeptid is not only hydrophilic but also lipophilic amphipathic molecule, it is powerful tensio-active agent, can greatly reduce surface tension and change surface property, thereby aspect environmental pollution improvement, particularly have great application potential aspect the insoluble pollutent biological restoration such as oil and agricultural chemicals.Have been found that genus bacillus can produce three class lipopeptid class secondary metabolites: the plain class of surfactivity (Surfactins) is to be seven peptides of LLDLLDL and beta-hydroxy fatty acid (ring texture of fatty chain length C13～C15) form by lactone bond by a chirality; The plain class of her withered grass (Iturins) is to be seven peptides of LDDLLDL and the beta-amino lipid acid (ring texture that fatty chain length C14～C17) (β AA) forms by a chirality; Decapeptide and a beta-hydroxy fatty acid (fatty chain length C15～C17) form ring texture that Feng Yuansu class (Fengycins) is is LDDDLDLLLL by a chirality by lactone bond.The chirality of this three major types lipopeptid is different with the 3d space structure, and performance is also variant, and this is the discrepant major reason of bio-surfactant intensity that why produces between different strain, and therefore efficient to produce the lipopeptid bacterial strain be a crucial job in screening.The bacterium lipopeptid promotes the biological restoration function except having, and can also suppress the pathogenic micro-organism growth, compares with the microbiotic of synthetic, has that side effect is little, the active high characteristics of target, belongs to a class new antibiotic.In addition, owing to nontoxic, also can be used as the additive of foods and cosmetics to human body.
Summary of the invention
Technical problem to be solved by this invention provides a kind of bacillus pumilus of efficient product lipopeptid, and the lipopeptid that is produced can significantly promote the biodegradation process of oil and/or cyhalothrin pesticide degradation bacteria.
The technical problem that the present invention also will solve provides the application of above-mentioned bacillus pumilus in the biodegradation process that promotes oil and/or cyhalothrin pesticide degradation bacteria.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
The inventor separates from the Shengli Oil Field oil-polluted soils of Shandong, screens the new bacterial strain of bacillus pumilus that obtains also preservation is Y8A, its called after bacillus pumilus (Bacillus pumilus) of classifying.
This bacterial strain is preserved in Wuhan University China typical culture collection center (being called for short CCTCC) at present; Address: Wuhan Wuhan University; Postcode: 430072; The numbering of registering on the books: CCTCC NO:M 209108; Preservation date: on May 18th, 2009.With this bacterium as producing bacterial strain.
CCTCC NO:M 209108 bacterial strains have following character:
1, morphological specificity:
Bacillus pumilus Y8A is shaft-like, and size is 1.5～1.7 μ m * 2.5～3.0 μ m, and Gram-positive can be moved, and the adnation flagellum is arranged, and produces gemma.Cultivate 48h for 30 ℃ on the LB solid plate, faint yellow, circular, opaque, edge unfairness, surperficial flat, moistening coarse that bacterium colony is.
2, physiological and biochemical property:
The physio-biochemical characteristics of bacillus pumilus Y8A see Table 1.
The physiological and biochemical property of table 1 bacterial strain Y8A
Annotate: "+": expression utilizes or is positive; "-": be negative reaction
3,16SrDNA sequential analysis
The 16S rDNA sequence length that records bacillus pumilus Y8A is 1471bp, specifically sees shown in the SEQ ID No.1.Institute's order-checking is listed in RDP database compare of analysis, chooses 16S rDNA sequence relevant in the database, adopt Bioedit, MEGA3.1 software carries out Phylogenetic Analysis to Y8A, the constructing system evolutionary tree.The result shows that bacterial strain Y8A and bacillus pumilus (Bacillus pumilus) homology reaches 100%, in conjunction with above physio-biochemical characteristics, Y8A is accredited as bacillus pumilus again, is specially bacillus pumilus Y8A (CCTCC NO:M 209108).
4, the characteristic of bio-surfactant:
(1) extraction of bio-surfactant and quantitative analysis
The bio-surfactant crude product that extracts the bacillus pumilus Y8A product that obtains is the yellow powder powder.The output of bacillus pumilus Y8A bio-surfactant is 5.05gL
-1
(2) qualitative analysis of tensio-active agent:
The thin-layer chromatography (TLC) of the bio-surfactant crude product of bacillus pumilus Y8A detects: point sample is not opened up layer on silica-gel plate, and product is red-purple behind the triketohydrindene hydrate chromogenic reagent, and what the preliminary judgement bacterial strain produced is the lipopeptid type biological surfactant.
The bio-surfactant infrared spectra that bacillus pumilus Y8A produces detects: on the FT-IR spectrogram, and 3285～3295cm
-1Be to shrink vibrational band, 3070～3075cm by the NH that the molecule interchain hydrogen bond causes
-1Be the flexible bands of a spectrum of NH that the intramolecular hydrogen bond of NH group causes, 1650～1655cm
-1, 1534～1535cm
-1Be respectively acid amides bands of a spectrum I and II.These characteristic absorbance show that this surfactant molecule contains peptide chain.2960～2870cm on the spectrogram
-1With 1455～1380cm
-1Two place's absorption peaks are C-H stretching vibrations from aliphatic carbon chain, 1225～1231cm
-1And 699cm
-1Be the charateristic avsorption band of lactone, show that this surfactant molecule contains fatty acid molecule.Therefore, the bio-surfactant of bacillus pumilus Y8A generation belongs to the lipopeptid class.
(3) the fermented liquid emulsifying property is measured:
Fermented liquid with after paraffin mixes, is detected the variation of emulsification volume every 12h, and continuous monitoring 48h, the fermented liquid of bacterial strain Y8A be good emulsified olefin all.The Y8A fermented liquid is in 12h, and the emulsification volume maintains 100%.Along with the prolongation of time, the emulsification volume descends to some extent, but lowering speed is very slow, and the Y8A fermented liquid still remains on 80% at 48h internal emulsification volume.After placing 7d, the emulsification volume still can reach 70%.Therefore, bacillus pumilus Y8A fermented liquid emulsifying property is stable, and solubilizing effect is good.
(4) mensuration of micelle-forming concentration (CMC):
The tensio-active agent crude product that extracts is diluted to 10mgL respectively
-1, 20mgL
-1, 30mgL
-1, 40mgL
-1, 50mgL
-1, 100mgL
-1, 200mgL
-1After, the surface tension of the surfactant soln of survey different mass concentration.Along with the increase of tensio-active agent mass concentration, the surface tension of solution reduces gradually, drops to 23.4mNm
-1After remain unchanged substantially, calculating the CMC value that bacterial strain produces the thing tensio-active agent is 30mgL
-1, than CMC value (the SDS 2120mgL of general chemical surfactant
-1, polysorbas20 600mgL
-1) low 1～2 order of magnitude, so bio-surfactant is more superior at aspect of performance than general chemical surfactant.
5, fermention medium and Optimizing Conditions of Fermentation:
(1) the capillary relation of thalli growth and fermented liquid:
The increment of bacterial strain and fermented liquid surface tension are inverse ratio, and surface tension descends along with thalli growth, and when cell concentration reached maximum, it is minimum that surface tension reaches, and in whole stationary phase, the surface tension variations of fermented liquid is little.Be 8h the lag period of bacterial strain Y8A growth, enters logarithmic phase behind the 8h, reaches maximum at 22 h cell concentrations, and it is minimum that the surface tension of nutrient solution correspondingly reaches.
(2) different carbon sources are to the thalli growth and the influence of producing tensio-active agent:
Add different carbon sources (sucrose, glucose, lactose, wood sugar, maltose) in the LB substratum, final concentration is 10gL
-1Bacterial strain Y8A is equal well-grown in above-mentioned carbon source, can obviously reduce bacterial strain Y8A fermented liquid surface tension, the minimum 28.6mNm that reaches but add lactose
-1
(3) temperature influence that thalli growth and tensio-active agent are produced:
Culture temperature is chosen 20 ℃ respectively, 25 ℃, 30 ℃, 37 ℃, does not change other fermentation condition.30 ℃ is the optimum growth temperature of bacterial strain Y8A, the amount of surfactant maximum of generation, the minimum 28.3mNm that reaches of fermented liquid surface tension value
-1But the generation of thalli growth and tensio-active agent is also all better when 20 ℃ and 25 ℃, shows that it has the thermal adaptation scope of broad.And 37 ℃ of growth effects to thalline are little, but very big to exerting an influence of tensio-active agent.Only illustrate that in certain temperature range, the growth of bacterial strain Y8A and the temperature of synthetic surfactant are corresponding to.
(4) pH value influence that thalli growth and tensio-active agent are produced:
The initial pH of fermention medium is transferred to 4,5,5.5,6,6.5,7,7.5,8,8.5,9,10,11,12 respectively, and other fermentation condition does not change.Bacterial strain Y8A grows all good in the pH value is 5～12 scopes, and the tensio-active agent of generation makes the surface tension of nutrient solution that remarkable reduction be arranged, and difference is little, but when pH4.0, growth seriously is suppressed.Illustrate that Y8A has very wide pH subject range.
(5) air flow influence that thalli growth and tensio-active agent are produced:
In the 250mL triangular flask, add 25,50,75,100,125 respectively, the 150mL fermention medium, other fermentation condition does not change.Bacterial strain Y8A liquid amount for 50mL time growth best, the tension value of nutrient solution is minimum, and when other liquid amounts, the surface tension of growth and nutrient solution is slightly decline all.Illustrate that Y8A is less demanding to air flow, can be under the less condition of oxygen content fine growth and produce tensio-active agent.
(6) metal ion influence that thalli growth and tensio-active agent are produced:
In substratum, add each metal ion species (Fe respectively
2+, Cu
2+, Zn
2+, Mn
2+, Ca
2+), make its final concentration be 20mgL
-1Other fermentation condition does not change.20mgL
-1Ca
2+And Fe
2+Can significantly promote the growth of Y8A and the generation of tensio-active agent, the fermented liquid surface tension is reduced, and 20mgL
-1Zn
2+And Cu
2+Suppress the growth of Y8A and the generation of tensio-active agent.20mgL-1 Mn
2+The growth of Y8A and the generation of tensio-active agent there is slight promoter action.
(7) resistance of strains
With 1% inoculum size is 10mgL with the seed liquor access antibiotic concentration of bacterial strain Y8A respectively
-1The LB liquid nutrient medium in, 30 ℃, 170rpm are cultivated 24h.In nine kinds of microbiotic for examination, Y8A all has resistance except gentamicin is not had the resistance to other eight kinds of microbiotic, sees Table 2.
Table 2 bacterial strain Y8A is to antibiotic resistance
Annotate: "+" expression growth; "-" expression is not grown
To sum up, the culture condition that lipopeptide-producing bacillus pumilus of the present invention produces lipopeptid is: in the LB substratum, be carbon source with sucrose, glucose, lactose, wood sugar or maltose, add metal cations Fe
2+, Ca
2+Or Mn
2+, pH5～12 are cultured to cell concentration at 20～30 ℃ and reach maximum.Preferred culture condition is: in the LB substratum, be carbon source with the lactose, add metal cations Fe
2+Or Ca
2+, pH5～12 are cultured to cell concentration at 20～30 ℃ and reach maximum.Preferred culture condition is: in the LB substratum, be carbon source with the lactose, add metal cations Fe
2+Or Ca
2+, pH7.5 is cultured to cell concentration at 30 ℃ and reaches maximum, with this understanding, produces the lipopeptid ability and reaches the strongest.Wherein, described carbon source concentration is 10gL
-1, described concentration of metal ions is 20mgL
-1
Bacillus pumilus of the present invention (Bacillus pumilus) Y8A (CCTCC NO:M 209108), can efficiently produce the lipopeptid type biological surfactant, the site that contacts that tight adhesion is disperseed, increased organic pollutant degradation bacterium and pollutent at the strong-hydrophobicity organic pollutant on soil particle surface, therefore can unite use with other organic pollutant degradation bacterium such as oil and/or cyhalothrin pesticide degradation bacteria, accelerate the degradation process of oil and/or cyhalothrin agricultural chemicals.
Preferably, lipopeptide-producing bacillus pumilus Y8A of the present invention and the golden yellow Bacillaceae bacterial strain of alkane degradation bacterial and/or aromatic hydrocarbon degradation bacteria chlorophenol Sphingol single-cell bacterial strain co-inoculation in the soil or waters of petroleum pollution, are promoted the microbiological deterioration process of oil.
Preferably, lipopeptide-producing bacillus pumilus Y8A of the present invention and cyhalothrin degradation bacteria bacillus cereus are belonged to the bacterial strain co-inoculation in the soil or waters of cyhalothrin pesticidal contamination, promote cyhalothrin microorganism Decompounding of Pesticide process.
Beneficial effect: the strain of i (bacillus) pumilus Y8A that the present invention screens (CCTCC NO:M 209108), generation lipopeptid type biological surfactant that can efficient stable can effectively promote the biodegradation process of oil and/or cyhalothrin pesticide degradation bacteria.The efficient product lipopeptid function of bacterial strain Y8A itself is expected to be widely used in agricultural, medicine, makeup and food and chemical industry as production of antibiotics bacterium of new generation and chaotropic agent, among other organic contamination degraded microorganism can be used for the contaminate environment reparation.The multiple microbiotic of this bacterial strain ability, fermenting process is simple, processing ease.
Description of drawings
The negative staining electron microscope photo of Fig. 1 bacillus pumilus Y8A of the present invention.
The haemolysis situation (left side) of Fig. 2 bacillus pumilus Y8A of the present invention on blood agar, lipopeptid Acinetobacter bacterial strain [Acinetobacter sp.BHSN (EU293155)] is produced in another strain of arriving of this laboratory separation screening in contrast.
The emulsifying effectiveness (left side) of Fig. 3 bacillus pumilus Y8A of the present invention fermented liquid, lipopeptid Acinetobacter bacterial strain [Acinetobacter sp.BHSN (EU293155)] is produced in another strain of arriving of this laboratory separation screening in contrast.
The infrared absorpting light spectra of the bio-surfactant that Fig. 4 bacillus pumilus Y8A of the present invention produces.
Embodiment:
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand that the described concrete material proportion of embodiment, processing condition and result thereof only are used to illustrate the present invention, and should also can not limit the present invention described in detail in claims.
Embodiment 1:
In the LB substratum, add 20mgL
-1Fe
2+And Ca
2+Can significantly promote the growth of Y8A and the generation of tensio-active agent; 10gL
-1Lactose can significantly promote the generation of tensio-active agent; Bacterial strain Y8A is at 20～30 ℃, and the ability of growth and generation tensio-active agent is all stronger in pH5～12 scopes; At capacity is in the 250mL triangular flask, and liquid seeds is connected in the 50mL LB substratum by 1% inoculum size and (adds 10gL
-1Lactose, 20mgL
-1Fe
2+And Ca
2+, pH7.5), 30 ℃ of following 170rpm shaking tables are cultivated 24h.Measure the surface tension value of thalline fermented liquid after the fermentation ends, this is worth I and reaches 23.5mNm
-1
Embodiment 2:
Get the oil-polluted soils mixing that sieves after air-dry, 121 ℃ of sterilization 30min, oven dry.Be divided into six groups and compare test, every group of three of design is parallel: (A) add and produce lipopeptid bacterial strain Y8A bacterium liquid, (B) add the golden yellow Bacillaceae bacterial strain of alkane degradation bacterial (Chryseobacterium sp.ATCC 11642), (C) add aromatic hydrocarbon degradation bacteria chlorophenol Sphingol single-cell bacterial strain (Sphingomonas chlorophenolica ATCC 39723) bacterium liquid, (D) add Y8A and golden yellow Bacillaceae bacterial strain bacterium liquid, (E) add Y8A and chlorophenol Sphingol single-cell bacterium liquid, (F) add Y8A, golden yellow Bacillaceae bacterial strain and chlorophenol Sphingol single-cell bacterium liquid.Connect the bacterium amount and be 1.1 * 10
8Individual g
-1, mixing is contrast (CK) not add bacterium liquid soil.The vessel surface covered with plastic film is pricked the hole ventilation, cultivates 30d for 30 ℃, adopts crude oil residual quantity in the gravimetric determination soil.Test-results such as table 3.
Table 3 Y8A is to the promoter action of petroleum pollution biological restoration
Embodiment 3:
(final concentration is 100mgL to contain cyhalothrin at 1/10LB
-1) inserting cyhalothrin degradation bacteria bacillus cereus strain (Bacillus cereus ATCC7039) and product lipopeptid bacterial strain Y8A of the present invention simultaneously in the substratum, inoculative proportion is 1: 1, inoculum size is 1%, measures fermented liquid OD respectively at different time
600With the cyhalothrin residual amount.During with the bacillus cereus strain single culture, the growth of thalline with the cyhalothrin degradation rate is compared, during mixed culture, the cyhalothrin degradation speed is obviously accelerated, reached the most degradation amount in 34 hours, do not shifted to an earlier date 7 hours than adding Y8A, degradation process has shortened nearly 1/6.The measurement result of cultivating after 2 days shows, when cyhalothrin degradation bacteria bacillus cereus single culture, the degradation rate of cyhalothrin is 38.7%, when bacillus cereus and Y8A co-cultivation, the degradation rate of cyhalothrin improves significantly to 87.7%, so Y8A can promote the biological degradation of agricultural chemicals significantly.
SEQUENCE?LISTING
＜110〉Agricultural University Of Nanjing
＜120〉a kind of lipopeptide-producing bacillus pumilus and application thereof
<130>njau090521
<160>1
<170>PatentIn?version 3.3
<210>1
<211>1471
<212>DNA
＜213〉bacillus pumilus (Bacillus pumilus)
<400>1
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ccggatgtta?gcggcggacg?ggtgagtaac?acgtgggtaa?cctgcctgta?agactgggat 120
aactccggga?aaccggagct?aataccggat?agttccttga?accgcatggt?tcaaggatga 180
aagacggttt?cggctgtcac?ttacagatgg?acccgcggcg?cattagctag?ttggtggggt 240
aatggctcac?caaggcgacg?atgcgtagcc?gacctgagag?ggtgatcggc?cacactggga 300
ctgagacacg?gcccagactc?ctacgggagg?cagcagtagg?gaatcttccg?caatggacga 360
aagtctgacg?gagcaacgcc?gcgtgagtga?tgaaggtttt?cggatcgtaa?agctctgttg 420
ttagggaaga?acaagtgcga?gagtaactgc?tcgcaccttg?acggtaccta?accagaaagc 480
cacggctaac?tacgtgccag?cagccgcggt?aatacgtagg?tggcaagcgt?tgtccggaat 540
tattgggcgt?aaagggctcg?caggcggttt?cttaagtctg?atgtgaaagc?ccccggctca 600
accggggagg?gtcattggaa?actgggaaac?ttgagtgcag?aagaggagag?tggaattcca 660
cgtgtagcgg?tgaaatgcgt?agagatgtgg?aggaacacca?gtggcgaagg?cgactctctg 720
gtctgtaact?gacgctgagg?agcgaaagcg?tggggagcga?acaggattag?ataccctggt 780
agtccacgcc?gtaaacgatg?agtgctaagt?gttagggggt?ttccgcccct?tagtgctgca 840
gctaacgcat?taagcactcc?gcctggggag?tacggtcgca?agactgaaac?tcaaaggaat 900
tgacgggggc?ccgcacaagc?ggtggagcat?gtggtttaat?tcgaagcaac?gcgaagaacc 960
ttaccaggtc?ttgacatcct?ctgacaaccc?tagagatagg?gctttccctt?cggggacaga 1020
gtgacaggtg?gtgcatggtt?gtcgtcagct?cgtgtcgtga?gatgttgggt?taagtcccgc 1080
aacgagcgca?acccttgatc?ttagttgcca?gcattcagtt?gggcactcta?aggtgactgc 1140
cggtgacaaa?ccggaggaag?gtggggatga?cgtcaaatca?tcatgcccct?tatgacctgg 1200
gctacacacg?tgctacaatg?gacagaacaa?agggctgcaa?gaccgcaagg?tttagccaat 1260
cccataaatc?tgttctcagt?tcggatcgca?gtctgcaact?cgactgcgtg?aagctggaat 1320
cgctagtaat?cgcggatcag?catgccgcgg?tgaatacgtt?cccgggcctt?gtacacaccg 1380
cccgtcacac?cacgagagtt?tgcaacaccc?gaagtcggtg?aggtaacctt?tatggagcca 1440
gccgccgaag?gtggggcaga?tgattggggt?g 1471
Claims (9)
1, a kind of lipopeptide-producing bacillus pumilus, its classification called after bacillus pumilus (Bacillus pumilus) Y8A has been preserved in Wuhan University China typical culture collection center, and its deposit number is CCTCC NO:M 209108.
2, the application of the described lipopeptide-producing bacillus pumilus of claim 1 in promoting oil and/or cyhalothrin microbe degradation of pesticides process.
3, application according to claim 2 is characterized in that the culture condition of lipopeptide-producing bacillus pumilus is: in the LB substratum, be carbon source with sucrose, glucose, lactose, wood sugar or maltose, add metal cations Fe
2+, Ca
2+Or Mn
2+, pH5～12 are cultured to cell concentration at 20～30 ℃ and reach maximum.
4, application according to claim 3 is characterized in that the culture condition of lipopeptide-producing bacillus pumilus is: in the LB substratum, be carbon source with the lactose, add metal cations Fe
2+Or Ca
2+, pH5～12 are cultured to cell concentration at 20～30 ℃ and reach maximum.
5, application according to claim 4 is characterized in that the culture condition of lipopeptide-producing bacillus pumilus is: in the LB substratum, be carbon source with the lactose, add metal cations Fe
2+Or Ca
2+, pH7.5 is cultured to cell concentration at 30 ℃ and reaches maximum.
6,, it is characterized in that described carbon source concentration is 10gL according to any described application among the claim 3-5
-1
7,, it is characterized in that described concentration of metal ions is 20mgL according to any described application among the claim 3-5
-1
8, application according to claim 2, it is characterized in that, lipopeptide-producing bacillus pumilus Y8A and the golden yellow Bacillaceae bacterial strain of alkane degradation bacterial and/or aromatic hydrocarbon degradation bacteria chlorophenol Sphingol single-cell bacterial strain co-inoculation in the soil or waters of petroleum pollution, are promoted the microbiological deterioration process of oil.
9, application according to claim 2, it is characterized in that, lipopeptide-producing bacillus pumilus Y8A and cyhalothrin degradation bacteria bacillus cereus are belonged to the bacterial strain co-inoculation in the soil or waters of cyhalothrin pesticidal contamination, promote cyhalothrin microorganism Decompounding of Pesticide process.
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| CN108184836A (en) * | 2017-11-23 | 2018-06-22 | 北京联合大学 | A kind of combined fermentation object for accelerating crops degrading pesticide residue |
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Application publication date: 20091021 Assignee: HUAI'AN CHAIMIHE AGRICULTURAL SCIENCE AND TECHNOLOGY DEVELOPMENT CO.,LTD. Assignor: Nanjing Agricultural University Contract record no.: 2014320000173 Denomination of invention: Lipopeptide-producing bacillus pumilus and application thereof Granted publication date: 20101117 License type: Exclusive License Record date: 20140305 |
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