CN104774798A - Bacillus subtillis capable of effectively degrading diethylstilbestrol - Google Patents
Bacillus subtillis capable of effectively degrading diethylstilbestrol Download PDFInfo
- Publication number
- CN104774798A CN104774798A CN201510220160.1A CN201510220160A CN104774798A CN 104774798 A CN104774798 A CN 104774798A CN 201510220160 A CN201510220160 A CN 201510220160A CN 104774798 A CN104774798 A CN 104774798A
- Authority
- CN
- China
- Prior art keywords
- diethylstilbestrol
- des
- bacillus subtillis
- metabolizing
- bacterial strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
-
- 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
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
- A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
-
- A—HUMAN NECESSITIES
- A62—LIFE-SAVING; FIRE-FIGHTING
- A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
- A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
- A62D2101/20—Organic substances
- A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Toxicology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Business, Economics & Management (AREA)
- Emergency Management (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to the field of environmental management conducted through microorganism, in particular to bacillus subtillis capable of metabolizing diethylstilbestrol. The bacterial strain number of the bacillus subtillis capable of metabolizing diethylstilbestrol is JF. The bacillus subtillis capable of metabolizing diethylstilbestrol is registered and preserved in China General Microbiological Culture Collection Center (CGMCC) on July 22, 2013, and the preservation number is CGMCC NO. 7950. The bacillus subtillis capable of metabolizing diethylstilbestrol can effectively degrade diethylstilbestrol, corresponding environment-friendly biological agents can be developed through the bacterial strain, and the bacillus subtillis capable of metabolizing diethylstilbestrol has good application prospects.
Description
Technical field
The present invention relates to and utilize microorganism to carry out field of environmental improvement, be specifically related to the subtilis of a strain degradable stilboestrol.
Background technology
Stilboestrol (diethylstilbestrol, DES) is a kind of non steroidal estrogen class medicine of synthetic, is once used as the medicine improving breeding performonce fo animals.DES, as a kind of endocrine disrupter, can cause hepar damnification, grows precocious, transgenation and hormonal imbalance, the disease such as women with breast cancer, ovarian cancer.Because it has impact to Hormone system, have chronic toxicity to body, U.S. FDA and The Ministry of Agriculture of the People's Republic of China, MOA all provide against it and are used in animal food production.
The residue problem of current stilboestrol is still very serious, in the waste water of sewage work's discharge in area, Catalonia, the content of DES is 34 ng/L, the high pavilions that relax etc. are randomly drawed in 40 increment product of detection the commercially available chicken in Nanyang and have all been detected DES, and what residual quantity was the highest reaches 10 mg/kg.Animal cultivation family Misuse, chemistry discharge, sewage work and refuse landfill etc. all may cause DES to residue in animal food, water body and edatope.In recent years, the method for degraded or elimination DES is subject to extensive concern.
Microbial degradation method is considered to the effective ways eliminating hazardous compound pollution, and the functional microorganism of screening efficient degradation DES is the effective ways realizing reducing or eliminating DES in environment.
Summary of the invention
The object of this invention is to provide a strain can effectively degrade stilboestrol bacterial strain-subtilis (
bacillus subtilis) JF.
Degradation bacteria strains provided by the present invention be subtilis (
bacillus subtilis) JF, be preserved in China General Microbiological culture presevation administrative center, preservation place is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and preserving number is CGMCC No. 7950, and preservation date is on July 22nd, 2013.
The step of described subtilis degraded DES is: 1) bacterial strain JF activates: picking subtilis JF rules on LB substratum, cultivates 24 h for 30 DEG C.
2) bacterial strain JF degrade DES: with stroke-physiological saline solution by the lawn wash-out on LB substratum and mixing prepare bacteria suspension.Pipette 1 mL bacterial suspension inoculation in the 250 mL triangular flasks that 29 mL LB substratum (containing 0.2% tween 80) are housed, add DES to finite concentration, replace bacteria suspension for blank with 1 mL physiological saline.30 DEG C, 180 r/min shaking culture, timing sampling, extracts DES, adopts HPLC to measure DES degradation rate.
In described microbiological deterioration DES system, the extracting method of DES is: accurately pipette the even nutrient solution of 1 mL in 10 mL scale test tubes, with methanol constant volume to 10 mL, and supersound extraction 20 min in ultrasonic cleaner (AS10200A); Mixing, gets centrifugal 15 min of 1.5 mL sample 12 000 r/min, by supernatant liquor 0.45 μm of organic phase membrane filtration, discard just filtrate about 0.5 mL, collection subsequent filtrate is sample solution, and analyze for HPLC, analytical instrument is LC-10A2010C HT type high performance liquid chromatograph.
In described microbiological deterioration system, DES detection method is: LC-10A2010C HT type liquid chromatographic system, and Sepax GP-C18 post (150 mm × 4.60 mm, 5.0 μm) is chromatographic column, methyl alcohol: water (7:3; V/v) be moving phase, flow velocity 0.7 mL/min, sample size 10 μ L, detects at 242 nm places by UV-detector.
Used medium of the present invention is LB substratum: yeast extract paste 5 g, peptone 10 g, sodium-chlor 10 g, water 1000 mL, regulates pH to 7.0,121 DEG C of sterilizing 15 min.
Compared with prior art, positively effect of the present invention is: the bacterial classification that subtilis belongs to " generally recognized as safe ", this subtilis degraded DES, there is the advantages such as degradation speed is fast, expense is low, operation steps is simple, provide good bacterium source and thinking for the DES thoroughly removed in environment remains.
Accompanying drawing explanation
Fig. 1 is the HPLC spectrogram of DES before and after degradation bacteria strains JF process, in figure, 0 h samples the color atlas processing the HPLC detection obtained at once in the LB substratum of the 25 mg/L DES containing inoculation JF, and 5 d sample the color atlas processing the HPLC detection obtained after cultivation 5 d.
Fig. 2 is the degradation curve that degradation bacteria strains JF acts on 25 mg/L DES.
Fig. 3 is the degradation capability of degradation bacteria strains JF to different concns DES.
Embodiment
Example 1: different incubation time is to the situation of bacterial strain JF degraded DES.
Prepared by seed liquor: picking JF bacterial classification is in the flat lining out of LB, and cultivate 24 h in 30 DEG C of constant incubators after, picking list bacterium colony is rule on LB slant medium, cultivates 24 h in 30 DEG C of constant incubators; With stroke-physiological saline solution by the JF lawn wash-out on slant medium and mixing make seed liquor (bacteria concentration be about 10
8cFU/mL).
Get 1 mL seed liquor be inoculated in 250 mL triangular flasks be equipped with 29 mL DES concentration be 25 mg/L containing 0.2% tween 80 LB substratum in, replace seed liquor for blank with 1 mL physiological saline, 30 DEG C, 180 r/min shaking culture 9 d, every 24 h samplings, Extraction solvent is made with methyl alcohol, supersound extraction, HPLC detects DES concentration.Before and after degradation bacteria strains JF process, the HPLC spectrogram of DES as shown in Figure 1; Degradation bacteria strains JF acts on the degradation curve of 25 mg/L DES as shown in Figure 2.Bacterial strain JF has obvious Degradation to DES, can degradable 25 mg/L DES in 9 d.
Described DES extracting method is: accurately pipette the even nutrient solution of 1 mL in 10 mL scale test tubes, with methanol constant volume to 10 mL, and supersound extraction 20 min in ultrasonic cleaner (AS10200A); Mixing, gets centrifugal 15 min of 1.5 mL sample 12 000 r/min, by supernatant liquor 0.45 μm of organic phase membrane filtration, discard just filtrate about 0.5 mL, collection subsequent filtrate is sample solution, and analyze for HPLC, detecting instrument is LC-10A2010C HT type high performance liquid chromatograph.
Described HPLC detection method is: LC-10A2010C HT type liquid chromatographic system, Sepax GP-C18 post (150 mm × 4.60 mm, 5.0 μm) be chromatographic column, methyl alcohol: water (7:3 v/v) is moving phase, flow velocity 0.7 mL/min, sample size 10 μ L, detects at 242 nm places by UV-detector.
The calculation formula of DES degradation rate is:
In formula: C
0for DES concentration (mg/L) in blank; C is DES residual concentration (mg/L) in nutrient solution.
Described substratum is: LB substratum: yeast extract paste 5 g, peptone 10 g, sodium-chlor 10 g, water 1000 mL, regulates pH to 7.0,121 DEG C of sterilizing 15 min.
Example 2: degradation bacteria strains JF is to the situation of degrading of different concns DES.
Get 1 mL seed liquor be inoculated in 250 mL triangular flasks be equipped with 29 mL DES concentration be respectively 10,25,50,80 mg/L containing in the LB substratum of 0.2% tween 80, replace seed liquor for blank with 1 mL physiological saline, 30 DEG C, 180 r/min shaking culture 5 d, the 5th d respectively at 0 h and cultivation samples, methyl alcohol supersound extraction, HPLC detects DES concentration.Described DES extracting method and HPLC detection method are with implementing example 1.Degradation bacteria strains JF to the degraded situation of different concns DES as shown in Figure 3, bacterial strain JF all has Degradation to a certain degree to the DES of 10-80 mg/L, the DES of 10 mg/L can be degradable by bacterial strain JF in 5 d, and along with the increase of DES concentration, bacterial strain JF reduces the degradation rate of DES.
Above example is only be described embodiments of the present invention; and not scope of the present invention is limited; for a person skilled in the art; can improve above-mentioned explanation or be out of shape, but all these improve or are out of shape the protection domain that all should fall into claim of the present invention and determine.
Claims (2)
1. a bacillus subtilis (
bacillus subtilisstrain) JF, it is characterized in that being preserved in China General Microbiological culture presevation administrative center, preservation place is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and preserving number is CGMCC No. 7950, and preservation date is on July 22nd, 2013.
2. subtilis described in claim 1 (
bacillus subtilisstrain) JF, is characterized in that: stilboestrol of can effectively degrading, for the residue degrading of stilboestrol in environment or elimination provide good bacterium source.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510220160.1A CN104774798A (en) | 2015-05-04 | 2015-05-04 | Bacillus subtillis capable of effectively degrading diethylstilbestrol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510220160.1A CN104774798A (en) | 2015-05-04 | 2015-05-04 | Bacillus subtillis capable of effectively degrading diethylstilbestrol |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104774798A true CN104774798A (en) | 2015-07-15 |
Family
ID=53616574
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510220160.1A Pending CN104774798A (en) | 2015-05-04 | 2015-05-04 | Bacillus subtillis capable of effectively degrading diethylstilbestrol |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104774798A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102911890A (en) * | 2011-08-05 | 2013-02-06 | 烟台海上传奇生物科技有限公司 | Pseudomonas capable of metabolizing diethylstilbestrol and application thereof |
| CN103421701A (en) * | 2012-05-24 | 2013-12-04 | 中国科学院生态环境研究中心 | Bacillus subtilis strain and application thereof to degradation of 17 beta-estradiol |
| CN103571771A (en) * | 2013-09-12 | 2014-02-12 | 四川农业大学 | Screening and identification and application of bacillus for efficiently degrading phthalate |
| CN104195084A (en) * | 2014-09-01 | 2014-12-10 | 南京农业大学 | Diethylstilbestrol degradation strain and application thereof in sewage and animal manure treatment |
-
2015
- 2015-05-04 CN CN201510220160.1A patent/CN104774798A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102911890A (en) * | 2011-08-05 | 2013-02-06 | 烟台海上传奇生物科技有限公司 | Pseudomonas capable of metabolizing diethylstilbestrol and application thereof |
| CN103421701A (en) * | 2012-05-24 | 2013-12-04 | 中国科学院生态环境研究中心 | Bacillus subtilis strain and application thereof to degradation of 17 beta-estradiol |
| CN103571771A (en) * | 2013-09-12 | 2014-02-12 | 四川农业大学 | Screening and identification and application of bacillus for efficiently degrading phthalate |
| CN104195084A (en) * | 2014-09-01 | 2014-12-10 | 南京农业大学 | Diethylstilbestrol degradation strain and application thereof in sewage and animal manure treatment |
Non-Patent Citations (2)
| Title |
|---|
| ZHANG WEIWEI ET AL.: "ISOLATION AND CHARACTERIZATION OF PSEUDOMONAS SP. STRAIN CAPABLE OF DEGRADING DIETHYLSTILBESTROL", 《APPLIED MICROBIOLOGY AND BIOTECHNOLOGY》 * |
| 杨俊等: "1株17β-雌二醇高效降解菌的分离鉴定及降解特性", 《环境科学》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103952362B (en) | One strain is to the oranges and tangerines endogeny rayungus of various plants pathogenic bacteria tool bacteriostatic activity | |
| CN101597578B (en) | Enramycin producing strain and extraction method by using macroporous resin | |
| CN101392227A (en) | Bacillus prodigiosus and prodigiosin producted thereby | |
| CN102311981B (en) | Method for preparing and purifying prodigiosin | |
| CN103695358B (en) | The one small streptomycete in strain ocean and the application in quorum sensing suppression thereof | |
| CN105255775A (en) | Solimonas capable of efficiently degrading aflatoxin B1 and application thereof | |
| CN105838614B (en) | A method of utilizing black-koji mould degrading zearalenone | |
| CN104531582A (en) | Strain for degrading pendimethalin pesticide and fungicide and application of strain | |
| CN111909881A (en) | Bacillus pumilus capable of producing feruloyl esterase and application thereof | |
| CN104830699A (en) | Aspergillus niger for effectively degrading diethylstibestrol | |
| CN104388319A (en) | Extraction method for citrus brown blotch germ crude toxin and toxin production condition | |
| CN109439588B (en) | Klebsiella strain, pyridine biodegradable microbial inoculum and preparation method and application thereof | |
| CN104004693B (en) | A kind of Bacillus pumilus and the application of earthy in controlling Chinese liquor thereof | |
| CN104774798A (en) | Bacillus subtillis capable of effectively degrading diethylstilbestrol | |
| Camacho et al. | Isolation and identification of a Bacillus sp. from freshwater sediment displaying potent activity against bacteria and phytopathogen fungi | |
| CN105420119A (en) | Ginseng endophytic fungus and application thereof | |
| CN104109642A (en) | Serratia marcescens, and screening method and application thereof | |
| CN110511876B (en) | Epimedium koreanum endophyte, culture method and metabolite thereof | |
| CN104804071A (en) | Depsipeptide compound, and preparation method and application thereof | |
| CN105352902B (en) | A kind of method of myriocin content in quick measure cicada fungus sample | |
| CN104004692B (en) | A kind of bacillus subtilis and the application of earthy in controlling Chinese liquor thereof | |
| CN105349465A (en) | Intertidal Exiguobacterium aestuarii and application thereof | |
| CN110885771A (en) | Biological agent for improving tobacco-planting soil micro-ecological environment level | |
| CN104388352A (en) | Pseudomonas stutzeri capable of simultaneously degrading chlorpyrifos and naphthalene | |
| CN104946555B (en) | A kind of microbacterium of degradable zeranol and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150715 |