CN110129226A - A kind of lysine bacillus and its acquisition methods and application - Google Patents
A kind of lysine bacillus and its acquisition methods and application Download PDFInfo
- Publication number
- CN110129226A CN110129226A CN201910405256.3A CN201910405256A CN110129226A CN 110129226 A CN110129226 A CN 110129226A CN 201910405256 A CN201910405256 A CN 201910405256A CN 110129226 A CN110129226 A CN 110129226A
- Authority
- CN
- China
- Prior art keywords
- lysine bacillus
- bacillus
- acquisition methods
- lysine
- shuttle shape
- 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.)
- Granted
Links
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 55
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 title claims abstract description 52
- 239000004472 Lysine Substances 0.000 title claims abstract description 52
- 238000000034 method Methods 0.000 title claims abstract description 21
- 229940011871 estrogen Drugs 0.000 claims abstract description 23
- 239000000262 estrogen Substances 0.000 claims abstract description 23
- 230000015556 catabolic process Effects 0.000 claims abstract description 21
- 238000006731 degradation reaction Methods 0.000 claims abstract description 21
- 150000000307 17β-estradiols Chemical class 0.000 claims abstract description 15
- 230000007613 environmental effect Effects 0.000 claims abstract description 15
- 241000894006 Bacteria Species 0.000 claims description 45
- 239000002609 medium Substances 0.000 claims description 41
- 239000000243 solution Substances 0.000 claims description 34
- 235000015097 nutrients Nutrition 0.000 claims description 23
- 239000012530 fluid Substances 0.000 claims description 21
- 239000006142 Luria-Bertani Agar Substances 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 8
- 239000012266 salt solution Substances 0.000 claims description 8
- 235000013619 trace mineral Nutrition 0.000 claims description 8
- 239000011573 trace mineral Substances 0.000 claims description 8
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 7
- 230000000813 microbial effect Effects 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 238000010790 dilution Methods 0.000 claims description 6
- 239000012895 dilution Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 6
- BFPYWIDHMRZLRN-SLHNCBLASA-N Ethinyl estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 BFPYWIDHMRZLRN-SLHNCBLASA-N 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 4
- 239000010802 sludge Substances 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 239000012137 tryptone Substances 0.000 claims description 4
- 229910021592 Copper(II) chloride Inorganic materials 0.000 claims description 3
- 241000726221 Gemma Species 0.000 claims description 3
- 239000007836 KH2PO4 Substances 0.000 claims description 3
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 claims description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 230000003647 oxidation Effects 0.000 claims description 3
- 238000007254 oxidation reaction Methods 0.000 claims description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 3
- 239000008223 sterile water Substances 0.000 claims description 3
- 239000011592 zinc chloride Substances 0.000 claims description 3
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 claims description 3
- PROQIPRRNZUXQM-UHFFFAOYSA-N (16alpha,17betaOH)-Estra-1,3,5(10)-triene-3,16,17-triol Natural products OC1=CC=C2C3CCC(C)(C(C(O)C4)O)C4C3CCC2=C1 PROQIPRRNZUXQM-UHFFFAOYSA-N 0.000 claims description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 2
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 claims description 2
- 229910004619 Na2MoO4 Inorganic materials 0.000 claims description 2
- 229910021586 Nickel(II) chloride Inorganic materials 0.000 claims description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- RGLYKWWBQGJZGM-ISLYRVAYSA-N diethylstilbestrol Chemical compound C=1C=C(O)C=CC=1C(/CC)=C(\CC)C1=CC=C(O)C=C1 RGLYKWWBQGJZGM-ISLYRVAYSA-N 0.000 claims description 2
- 229960000452 diethylstilbestrol Drugs 0.000 claims description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 2
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 2
- 229910052564 epsomite Inorganic materials 0.000 claims description 2
- 229960001348 estriol Drugs 0.000 claims description 2
- PROQIPRRNZUXQM-ZXXIGWHRSA-N estriol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H]([C@H](O)C4)O)[C@@H]4[C@@H]3CCC2=C1 PROQIPRRNZUXQM-ZXXIGWHRSA-N 0.000 claims description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 2
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 2
- QMMRZOWCJAIUJA-UHFFFAOYSA-L nickel dichloride Chemical compound Cl[Ni]Cl QMMRZOWCJAIUJA-UHFFFAOYSA-L 0.000 claims description 2
- 239000011684 sodium molybdate Substances 0.000 claims description 2
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 150000002576 ketones Chemical class 0.000 claims 1
- DNXHEGUUPJUMQT-CBZIJGRNSA-N Estrone Chemical compound OC1=CC=C2[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1 DNXHEGUUPJUMQT-CBZIJGRNSA-N 0.000 abstract description 27
- DNXHEGUUPJUMQT-UHFFFAOYSA-N (+)-estrone Natural products OC1=CC=C2C3CCC(C)(C(CC4)=O)C4C3CCC2=C1 DNXHEGUUPJUMQT-UHFFFAOYSA-N 0.000 abstract description 14
- 229960003399 estrone Drugs 0.000 abstract description 14
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 abstract description 13
- 229960005309 estradiol Drugs 0.000 abstract description 9
- 238000004321 preservation Methods 0.000 abstract description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 5
- 229910052799 carbon Inorganic materials 0.000 abstract description 5
- 229930182833 estradiol Natural products 0.000 abstract description 5
- 239000001963 growth medium Substances 0.000 abstract description 5
- 230000000593 degrading effect Effects 0.000 abstract description 2
- 238000012545 processing Methods 0.000 abstract description 2
- 238000005067 remediation Methods 0.000 abstract description 2
- 230000001580 bacterial effect Effects 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- IISBACLAFKSPIT-UHFFFAOYSA-N bisphenol A Chemical compound C=1C=C(O)C=CC=1C(C)(C)C1=CC=C(O)C=C1 IISBACLAFKSPIT-UHFFFAOYSA-N 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000008055 phosphate buffer solution Substances 0.000 description 3
- NTXOYFCYRZDHIN-UHFFFAOYSA-N 1,2,2,2-tetrachloroethylbenzene Chemical class ClC(Cl)(Cl)C(Cl)C1=CC=CC=C1 NTXOYFCYRZDHIN-UHFFFAOYSA-N 0.000 description 2
- 241001468185 Caryophanon Species 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 241001472591 Lysinibacillus sp. Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 229940106691 bisphenol a Drugs 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- BFPYWIDHMRZLRN-UHFFFAOYSA-N 17alpha-ethynyl estradiol Natural products OC1=CC=C2C3CCC(C)(C(CC4)(O)C#C)C4C3CCC2=C1 BFPYWIDHMRZLRN-UHFFFAOYSA-N 0.000 description 1
- DUZCMUMEHMNJCX-HAMGIEOISA-N C=1C=C(O)C=CC=1C(/CC)=C(\CC)C1=CC=C(O)C=C1.C=1C=C(O)C=CC=1C(/CC)=C(\CC)C1=CC=C(O)C=C1 Chemical compound C=1C=C(O)C=CC=1C(/CC)=C(\CC)C1=CC=C(O)C=C1.C=1C=C(O)C=CC=1C(/CC)=C(\CC)C1=CC=C(O)C=C1 DUZCMUMEHMNJCX-HAMGIEOISA-N 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- -1 E2) Chemical compound 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000568397 Lysinibacillus Species 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229910015667 MoO4 Inorganic materials 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- BHLCQSSLVPEHCW-BTVCFUMJSA-N [P].O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO Chemical compound [P].O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO BHLCQSSLVPEHCW-BTVCFUMJSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
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- 230000009471 action Effects 0.000 description 1
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 238000011953 bioanalysis Methods 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
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- 239000006227 byproduct Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
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- 235000009508 confectionery Nutrition 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000598 endocrine disruptor Substances 0.000 description 1
- 210000000750 endocrine system Anatomy 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000001076 estrogenic effect Effects 0.000 description 1
- 229960002568 ethinylestradiol Drugs 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 230000008717 functional decline Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 238000001728 nano-filtration Methods 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 210000004994 reproductive system Anatomy 0.000 description 1
- 208000017443 reproductive system disease Diseases 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001223 reverse osmosis Methods 0.000 description 1
- 238000009938 salting Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000032678 sex differentiation Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- 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
-
- 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
- C12N1/02—Separating microorganisms from their culture media
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- 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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- 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
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- 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
- C02F2101/34—Organic compounds containing oxygen
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- 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
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- Life Sciences & Earth Sciences (AREA)
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- Bioinformatics & Cheminformatics (AREA)
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- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
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- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Water Supply & Treatment (AREA)
- Environmental & Geological Engineering (AREA)
- Hydrology & Water Resources (AREA)
- Biodiversity & Conservation Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of lysine bacillus and its acquisition methods and applications, belong to technical field of bioengineering.The lysine bacillus is named as shuttle shape lysine bacillus DH-B01, and in preservation on January 14 in 2019 to China typical culture collection center, deposit number is CCTCC M 2019039.The shuttle shape lysine bacillus that the present invention filters out has very strong degradation capability to environmental estrogens such as estradiol and oestrone；Wherein, shuttle shape lysine bacillus DH-B01 culture is continuously cultivated 5 days in the culture medium using 17 beta estradiols or oestrone as sole carbon source, the estradiol or oestrone fast degradation that can be 30mg/L by concentration, degradation rate be up to 100%.Therefore the shuttle shape lysine bacillus DH-B01 can stablize, environmental estrogens of efficiently degrading, and can be applied to the biodegrade and environment remediation of processing environment estrogen.
Description
Technical field
The present invention relates to technical field of bioengineering, specifically a kind of lysine bacillus and its acquisition methods and answer
With.
Background technique
With the development of world economy, a kind of completely new environmental contaminants-incretion interferent (Endocrine
Disrupting Chemicals, i.e. EDCs) by common concern, have become after depletion of the ozone layer, greenhouse effects
Another global great environmental pollution problem seriously affects the survival and development of the mankind and the balance and stability of ecology.
With the understanding to incretion interferent, environmental estrogens (Environmental estrogen) are divided in environment
A major class important in chaff interferent is secreted, structurally and functionally similar to endogenous estrogen.This substance enters humans and animals
Analog estrogen action or change estrogenic activity, seriously affect aquatile Sex Differentiation, endanger to human health after in vivo
Evil is serious, damages to the nervous system, endocrine system, immune system of higher mammal, endangers maximum to reproductive system, makes
At male reproductive function decline and the reproductive diseases such as female acyesis and tumour.
With China's rapid economic development, no matter 17 beta estradiols (Estradiol, E2), oestrone (Estrone) etc. are natural
Estrogen, 17 α-ethinyl estradiol (Ethinylestradiol, EE2), diethylstilbestrol (Diethylstilbestrol) etc.
The industry such as artificial estrogen or bisphenol-A (Bisphenol A), double chlorphenyl trichloroethanes (DDT) pesticide oestrogen-like hormone all exists
Different water body environments detect, illustrate that the solution of environmental estrogens pollution problem is extremely urgent.
Currently, in the method for removal water body environment estrogen, based on the physical method, advanced oxidation either based on absorption
Chemical method, the further treatment technique even based on nanofiltration and reverse osmosis membrane technology, all because exist in water body environment it is a large amount of its
His competitive organic matter or self-defect are restricted in environmental estrogens pollutant removal.However based on biodegrade
Bioanalysis is a kind of important method for removing environment incretion interferent, since there is biological degradation method easy to operate, economy to have
The features such as imitating and not generating by-product, is with a wide range of applications.
About the project with biological method removal environmental estrogens, studies in China is started to walk relatively late, to microbial degradation
The mechanism study of environmental estrogens is less, and the degradation bacteria development and utilization of discovery are not thorough.Therefore, carry out and be directed to water body environment
The research and application of estrogen pollutant microorganism efficient degrading bacteria, to control and the pollution of Environment control hormone, regulation ecological system
System and safe drinking water are of great significance.
Summary of the invention
The purpose of the present invention is to provide a kind of lysine bacillus and its acquisition methods and applications, to solve above-mentioned back
The problem of being proposed in scape technology.
To achieve the above object, the invention provides the following technical scheme:
A kind of lysine bacillus is named as shuttle shape lysine bacillus DH-B01, lysine bacillus
(Lysinibacillus sp.), in preservation on January 14 in 2019 to China typical culture collection center, deposit number is
CCTCC M 2019039, preservation address are Wuhan City, Hubei Province Hongshan District Bayi Road Wuhan University.
Further, the 16S rDNA sequence of the lysine bacillus is as shown in SEQ ID NO:1 in sequence table.
The present invention also provides the acquisition methods of above-mentioned shuttle shape lysine bacillus DH-B01 a kind of, specifically, include with
Lower step:
(1) microbial source of the activated sludge in acquisition contraception pharmaceutical factory oxidation pond as the lysine bacillus；
(2) with the fluid nutrient medium containing 17 beta estradiols, inorganic salt solution and trace element solution to microbial source into
Row culture, obtains bacterium solution；
(3) bacterium solution obtained above is diluted to the dilution bacterium solution of various concentration gradient with sterile water, and by various concentration
The dilution bacterium solution of gradient is applied on LB agar medium and is cultivated, and then crosses, obtains on LB agar medium
The lysine bacillus.
Further, the mass concentration of 17 beta estradiols is 5-100mg/L in the fluid nutrient medium.
Further, the volume ratio of trace element solution and inorganic salt solution is 1:(95- in the fluid nutrient medium
105)；The pH of the fluid nutrient medium is 6.8-7.2.
Further, the inorganic salt solution is based on mass concentration including following components: Na2HPO4 4.2-4.3g/L、
KH2PO4 2.6-2.7g/L、MgSO4·7H2O 0.15-0.25g/L、(NH4)2SO4 1.4-1.5g/L、CaCl2 0.01-
0.03g/L。
Further, the trace element solution is based on mass concentration including following components: NiCl2·6H2O 0.02-
0.03g/L、CoCl2·6H2O 0.15-0.25g/L、H3BO3 0.005-0.01g/L、ZnCl2 0.05-0.1g/L、CuCl2·
2H2O 0.001-0.005g/L、MnSO4·H2O 0.06-0.07g/L、Na2MoO4·2H2O 0.02-0.03g/L。
Further, the LB agar medium is based on mass concentration including following components: tryptone 8-12g/L, ferment
Female extract 4-6g/L, NaCl 8-12g/L, agar powder 1-3g/L；The pH of the LB agar medium is 7.0-7.5.
The present invention also provides a kind of application of above-mentioned shuttle shape lysine bacillus DH-B01 in degradation environmental estrogens.
Further, the environmental estrogens include 17 beta estradiols, oestrone, estriol, 17 α-ethinyl estradiol and
Diethylstilbestrol.
Compared with prior art, the beneficial effects of the present invention are:
The present invention carries out acclimating, purifies and separates and secondary screening by the activated sludge in the pharmaceutical factory to production estrogen,
Obtain shuttle shape lysine bacillus DH-B01.The shuttle shape lysine bacillus that the present invention filters out is to estradiol and oestrone etc.
Environmental estrogens have very strong degradation capability；Wherein, shuttle shape lysine bacillus DH-B01 is cultivated with 17 β-female two
For alcohol continuously to cultivate 5 days in the culture medium of sole carbon source, shuttle shape is relied ammonia by the estradiol fast degradation that can be 30mg/L by concentration
Sour bacillus DH-B01 culture is continuously cultivated 5 days in the culture medium using oestrone as sole carbon source, can be 30mg/L by concentration
Oestrone fast degradation, the two degradation rate all reaches 100%.The shuttle shape lysine bacillus DH-B01 can stablize, efficiently
Degradation environmental estrogens, can be applied to the biodegrade and environment remediation of processing environment estrogen.
Detailed description of the invention
Fig. 1 is the shuttle shape lysine bacillus DH-B01 of the invention aspect graph on LB agar medium.
Fig. 2 is the aspect graph of shuttle shape lysine bacillus DH-B01 of the invention under an electron microscope.
Fig. 3 is the phylogenetic tree of shuttle shape lysine bacillus DH-B01 of the invention.
Fig. 4 is shuttle shape lysine bacillus DH-B01 of the invention to the degradation rate of 17 beta estradiols and the result of OD value
Figure.
Fig. 5 is shuttle shape lysine bacillus DH-B01 of the invention to the degradation rate of oestrone and the result figure of OD value.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
Belong to the scope of protection of the invention.
Embodiment 1
This embodiment offers the acquisition methods of shuttle shape lysine bacillus DH-B01 a kind of, specifically, including following
Step:
(1) microbial source of the activated sludge in acquisition contraception pharmaceutical factory as shuttle shape lysine bacillus DH-B01, and will
Laboratory is transported in its refrigeration back.
(2) it takes 17 beta estradiols of 25mg to be dissolved in the methanol of 50mL, it is female that 17 beta estradiols that concentration is 500mg/L is made
Liquid, and the 17 beta estradiol mother liquors of wherein 1mL is taken to be added in the triangular flask for filling 99mL fluid nutrient medium, make fluid nutrient medium
In 17 beta estradiols final concentration of 5mg/L；Then, triangular flask is put into thermostat water bath, water-bath at a temperature of 70 DEG C
30min；The methanol in fluid nutrient medium is set to volatilize completely；Then, height will be put into after triangular flask bottleneck gauze, brown paper sealing
Press autoclave in, make its 121 DEG C at a temperature of carry out high pressure sterilization 30min；After sterilizing, then the microbial source for taking 10ml to refrigerate
It is added in triangular flask, and triangular flask is placed in the constant-temperature shaking incubator of 30, DEG C 120rpm and carries out culture 72h, obtain just
Beginning bacterium solution.
(3) the 200 initial bacterium solutions of μ L are taken from the fluid nutrient medium of above-mentioned culture 72h, and fresh fluid nutrient medium is added,
Wherein 17 beta estradiol concentration are 10mg/L in the fluid nutrient medium；Then, which is placed in 30, DEG C 120rpm's
7d is continuously cultivated in constant-temperature shaking incubator, obtains secondary bacterium solution.
(4) 200 μ L, bis- bacterium solutions are taken to continuously add fresh fluid nutrient medium from the fluid nutrient medium of culture 7d, wherein
17 beta estradiol concentration are 20mg/L in the fluid nutrient medium；Then, by the fluid nutrient medium 30, the constant temperature of DEG C 120rpm shakes
It swings and continuously cultivates 7d in incubator；Then, the bacterium solution obtained after culture is inoculated into containing 17 β-female two using same method
Determining alcohol is respectively to obtain most in the fresh fluid nutrient medium of 40mg/L, 60mg/L, 80mg/L, 100mg/L (in triplicate)
Whole bacterium solution.
(5) the above-mentioned final bacterium solution of 1ml is taken, and (diluted concentration gradient is 10 with sterile water dilution-2~10-8)；Then,
It will be applied on LB agar medium by the dilution bacterium solution of different gradient dilutions, each gradient does 3 parallel groups；Then, will
LB agar medium is put into constant incubator, 30 DEG C at a temperature of cultivate 3d, observe bacterium colony growing way；Then, picking color,
The different bacterium colony such as form is repeatedly crossed on LB agar medium, until isolating single bacterium colony, is obtained shuttle shape and is relied ammonia
Sour bacillus DH-B01 bacterial strain, wherein the plate streaking figure of bacterial strain DH-B01 is as shown in Fig. 1.In addition, bacterial strain DH-B01 is passed through
After crossing expansion culture, a part can be placed in -80 DEG C of glycerol mixed liquor preservations, and a part can be placed in 4 DEG C of test tube slants and save.
The shuttle shape lysine bacillus DH-B01 that the embodiment obtains is on January 14th, 2019 in China typical culture collection
Heart preservation, preservation address are Wuhan City, Hubei Province Hongshan District Bayi Road Wuhan University, and number of registering on the books is CCTCC M
2019039。
It should be noted that above-mentioned fluid nutrient medium also contains inorganic salt solution and trace element solution, wherein nothing
It is respectively the Na of 4.26g/L containing mass concentration in machine salting liquid2HPO4, 2.65g/L KH2PO4, 0.2g/L MgSO4
(the NH of 7H2O, 1.5g/L4)2SO4With the CaCl of 0.02g/L2；Containing mass concentration in trace element solution is respectively 0.024g/
The NiCl of L2·6H2O, the CoCl of 0.19g/L2·6H2O, the H of 0.006g/L3BO3, 0.07g/L ZnCl2, 0.002g/L
CuCl2·2H2O, the MnSO of 0.061g/L4·H2The Na of O and 0.024g/L2MoO4·2H2O；Inorganic salt solution and microelement
The volume ratio of solution is 98:1, and is adjusted to the pH of fluid nutrient medium by addition 0.1mol/L NaOH and 0.1mol/L HCl
7.0。
In addition, above-mentioned LB agar medium contain mass concentration be respectively the tryptone of 10g/L, 5g/L yeast mention
Take NaCl the and 2g/L agar powder of object, 10g/L；When preparing LB agar medium, first by above-mentioned mass concentration by tryptone,
Yeast extract and NaCl are dissolved in water, and its pH is tuned into 7.4, then add agar thereto by above-mentioned mass concentration again
Powder can obtain LB agar medium.
Embodiment 2
The embodiment is identified the shuttle shape lysine bacillus DH-B01 bacterial strain of above-mentioned acquisition.Specifically, with
The universal primer of bacterium passes through PCR amplification DNA amplification.Sequencing result utilizes the BLAST tool and GenBank number in NCBI
Sequence analysis retrieval is carried out according to the 16S rDNA gene order in library.Comparison result shows shuttle shape lysine bacillus DH-
More bacillus in the 16S rDNA gene order of B01 and shuttle shape lysine bacillus (Lysinibacillus)
The gene order of 16S rDNA has higher homology.Choose the 16S rDNA base of 20 plants of bacteriums in lysine bacillus
It because of sequence, and chooses 1 plant of bacterial 16 S rDNA gene order in Caryophanon (Caryophanon) and is outer group, use MEGA
Software simultaneously carries out Phylogenetic analysis, the unrooted phylogenetic tree (such as attached drawing 3) of building, knot according to Neighbor-Joining
The morphological feature and physiological and biochemical index of bacillus DH-B01 are closed, Preliminary Identification bacterial strain DH-B01 is shuttle shape lysine gemma bar
Bacterium is named as shuttle shape lysine bacillus DH-B01.
In addition, the shuttle shape lysine bacillus DH-B01 of above-mentioned acquisition is on LB agar medium, bacterium colony is opaque, table
Face drying, neat in edge (such as attached drawing 1), observe the form of bacterial strain are as follows: thallus shape is in the shape of a rod, single bacterium under scanning electron microscope
About 2.0 × 1.0 μm of body size；Gemma is rounded or oval, diameter are greater than thallus, is located at thallus center, extreme or secondary extreme,
Make volumination in fusiform (such as attached drawing 2).Shuttle shape lysine bacillus DH-B01 glucose, nitrate (reduction), sweet dew
Alcohol, glucose phosphorus, maltose, sucrose detection are negative, with 3% hydrogen peroxide, semi-solid agar, lysozyme nutrient broth, matter
Control nutrient broth, urea tests positive.
Embodiment 3
The embodiment is degradation capability of the verifying shuttle shape lysine bacillus DH-B01 to 17 beta estradiols, specifically,
First by shuttle shape lysine bacillus DH-B01 strain inoculated into LB liquid medium, 48h is cultivated, bacterium solution is obtained；Then, will
Bacterium solution is poured into the centrifuge tube of sterilizing, and 10min is centrifuged under conditions of 7000rpm, abandons supernatant；Then, the phosphorus that pH is 7 is added
It after hydrochlorate buffer solution is washed, then is placed in the centrifuge tube of 7000rpm and carries out centrifugation 10min, remove supernatant.It is so anti-
After after backwashing washs 3 times, then thallus dispelled, it is 1.0 that bacterium solution, which is finally diluted to OD600 value, with phosphate buffer solution, and bacterium is made
Suspension is spare.Followed by the bacterium solution for taking out 3ml from bacteria suspension is inoculated into using 17 beta estradiols as sole carbon source, and concentration is
In the minimal medium of 30mg/L, 5d is continuously cultivated in 30, the constant-temperature shaking incubator of DEG C 120rpm, is taken daily primary
Sample.By culture medium after pretreatment, with high performance liquid chromatograph (High-performance liquid
Chromatography, HPLC) bacterium is detected to the degradation capability (testing result such as attached drawing 4) of 17 beta estradiols, to 17
The degradation rate of beta estradiol is up to 100%.Wherein, the condition of detection are as follows: detector UV (Dual λ Absorbance Detector,
Water2487), chromatographic column is Zorbax Eclipse Plus C18 column (150 × 4.6mm, 3.5mm)；Mobile phase volume ratio is
Acetonitrile: water=1:1, detector wavelength 275nm, flow velocity 0.8mL/min, sample volume are 10 μ L；With microplate reader detection bacterium
Upgrowth situation, the upgrowth situation of bacterium indicates with OD600.
Embodiment 4
The embodiment is to verify shuttle shape lysine bacillus DH-B01 to the degradation capability of oestrone, specifically, first by bacterium
Strain is inoculated into LB liquid medium, is cultivated 48h, is obtained bacterium solution；Then, bacterium solution is poured into the centrifuge tube of sterilizing,
It is centrifuged 10min under conditions of 7000rpm, abandons supernatant；Then the phosphate buffer solution that pH is 7 is added to be washed, then sets
Centrifugation 10min is carried out in the centrifuge tube of 7000rpm, removes supernatant.After washing 3 times repeatedly, then thallus dispelled, most
Afterwards, bacterium solution is diluted to OD600 value with phosphate buffer solution is 1.0, and it is spare that bacteria suspension is made.Followed by from bacteria suspension
The bacterium solution for taking out 3ml is inoculated into using oestrone as sole carbon source, and concentration is in the minimal medium of 30mg/L, 30, DEG C
5d is continuously cultivated in the constant-temperature shaking incubator of 120rpm, takes a sample daily.By culture medium after pretreatment, with efficient liquid
Chromatography detects the bacterium to the degradation capability (testing result such as attached drawing 5) of oestrone, to the degradation rate of oestrone up to 100%.
Wherein, the condition of detection are as follows: detector UV, chromatographic column be Zorbax Eclipse Plus C18 column (150 × 4.6mm,
3.5mm)；Mobile phase volume ratio is acetonitrile: water=1:1, detector wavelength 275nm, flow velocity 0.8mL/min, sample volume are
10μL；It is indicated with the upgrowth situation of the upgrowth situation of microplate reader detection bacterium, bacterium with OD600.
Taking the above-mentioned ideal embodiment according to the present invention as inspiration, through the above description, relevant staff is complete
Various changes and amendments can be carried out without departing from the scope of the technological thought of the present invention' entirely.The technology of this invention
Property range is not limited to the contents of the specification, it is necessary to which the technical scope thereof is determined according to the scope of the claim.
Sequence table
<110>Northeast Normal University
<120>a kind of lysine bacillus and its acquisition methods and application
<141> 2019-05-14
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1416
<212> DNA
<213>shuttle shape lysine bacillus (Lysinibacillus sp.)
<400> 1
gcaagtcgag cgaacagaga aggagcttgc tcctttgacg ttagcggcgg acgggtgagt 60
aacacgtggg caacctacct tatagtttgg gataactccg ggaaaccggg gctaataccg 120
aataatctgt ttcacctcat ggtgaaatat tgaaagacgg tttcggctgt cgctatagga 180
tgggcccgcg gcgcattagc tagttggtga ggtaacggct caccaaggcg acgatgcgta 240
gccgacctga gagggtgatc ggccacactg ggactgagac acggcccaga ctcctacggg 300
aggcagcagt agggaatctt ccacaatggg cgaaagcctg atggagcaac gccgcgtgag 360
tgaagaagga tttcggttcg taaaactctg ttgtaaggga agaacaagta cagtagtaac 420
tggctgtacc ttgacggtac cttattagaa agccacggct aactacgtgc cagcagccgc 480
ggtaatacgt aggtggcaag cgttgtccgg aattattggg cgtaaagcgc gcgcaggtgg 540
tttcttaagt ctgatgtgaa agcccacggc tcaaccgtgg agggtcattg gaaactggga 600
gacttgagtg cagaagagga tagtggaatt ccaagtgtag cggtgaaatg cgtagagatt 660
tggaggaaca ccagtggcga aggcgactat ctggtctgta actgacactg aggcgcgaaa 720
gcgtggggag caaacaggat tagataccct ggtagtccac gccgtaaacg atgagtgcta 780
agtgttaggg ggtttccgcc ccttagtgct gcagctaacg cattaagcac tccgcctggg 840
gagtacggtc gcaagactga aactcaaagg aattgacggg ggcccgcaca agcggtggag 900
catgtggttt aattcgaagc aacgcgaaga accttaccag gtcttgacat cccgttgacc 960
actgtagaga tatggtttcc ccttcggggg caacggtgac aggtggtgca tggttgtcgt 1020
cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct tgatcttagt 1080
tgccatcatt tagttgggca ctctaaggtg actgccggtg acaaaccgga ggaaggtggg 1140
gatgacgtca aatcatcatg ccccttatga cctgggctac acacgtgcta caatggacga 1200
tacaaacggt tgccaactcg cgagagggag ctaatccgat aaagtcgttc tcagttcgga 1260
ttgtaggctg caactcgcct acatgaagcc ggaatcgcta gtaatcgcgg atcagcatgc 1320
cgcggtgaat acgttcccgg gccttgtaca caccgcccgt cacaccacga gagtttgtaa 1380
cacccgaagt cggtgaggta acctttggag ccagcc 1416
Claims (10)
1. a kind of lysine bacillus, which is characterized in that the lysine bacillus is named as shuttle shape lysine gemma
Bacillus DH-B01, is preserved in China typical culture collection center, and deposit number is CCTCC M 2019039.
2. a kind of lysine bacillus according to claim 1, which is characterized in that the lysine bacillus
16S rDNA sequence is as shown in SEQ ID NO:1 in sequence table.
3. a kind of acquisition methods of such as lysine bacillus of any of claims 1-2, which is characterized in that including
Following steps:
(1) microbial source of the activated sludge in acquisition contraception pharmaceutical factory oxidation pond as the lysine bacillus；
(2) microbial source is trained with the fluid nutrient medium containing 17 beta estradiols, inorganic salt solution and trace element solution
It supports, obtains bacterium solution；
(3) bacterium solution obtained above is diluted to the dilution bacterium solution of various concentration gradient with sterile water, and by various concentration gradient
Dilution bacterium solution be applied on LB agar medium and cultivated, then cross, obtain described on LB agar medium
Lysine bacillus.
4. acquisition methods according to claim 3, which is characterized in that the matter of 17 beta estradiols in the fluid nutrient medium
Amount concentration is 5-100mg/L.
5. acquisition methods according to claim 3, which is characterized in that in the fluid nutrient medium trace element solution with
The volume ratio of inorganic salt solution is (0.5-1): 98；The pH of the fluid nutrient medium is 6.8-7.2.
6. acquisition methods according to claim 5, which is characterized in that the inorganic salt solution based on mass concentration including
Following components: Na2HPO4 4.2-4.3g/L、KH2PO4 2.6-2.7g/L、MgSO4·7H2O 0.15-0.25g/L、(NH4)2SO4 1.4-1.5g/L、CaCl2 0.01-0.03g/L。
7. acquisition methods according to claim 5, which is characterized in that the trace element solution wraps based on mass concentration
Include following components: NiCl2·6H2O 0.02-0.03g/L、CoCl2·6H2O 0.15-0.25g/L、H3BO3 0.005-0.01g/
L、ZnCl2 0.05-0.1g/L、CuCl2·2H2O 0.001-0.005g/L、MnSO4·H2O 0.06-0.07g/L、
Na2MoO4·2H2O 0.02-0.03g/L。
8. acquisition methods according to claim 3, which is characterized in that the LB agar medium wraps based on mass concentration
Include following components: tryptone 8-12g/L, yeast extract 4-6g/L, NaCl 8-12g/L, agar powder 1-3g/L；The LB
The pH of agar medium is 7.0-7.5.
9. a kind of application such as lysine bacillus of any of claims 1-2 in degradation environmental estrogens.
10. application according to claim 9, which is characterized in that the environmental estrogens include 17 beta estradiols, female
Ketone, estriol, 17 α-ethinyl estradiol and diethylstilbestrol.
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|---|---|---|---|---|
| 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 |
| CN107227266A (en) * | 2017-04-05 | 2017-10-03 | 东北师范大学 | A kind of Rhodococcus sp DSH of degradable environmental hormone bisphenol-A and its application |
| CN107523513A (en) * | 2017-04-05 | 2017-12-29 | 东北师范大学 | A kind of compound bacteria of 17 β estradiol capable of being fast degraded and its preparation method and application |
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2019
- 2019-05-14 CN CN201910405256.3A patent/CN110129226B/en not_active Expired - Fee Related
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| 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 |
| CN107227266A (en) * | 2017-04-05 | 2017-10-03 | 东北师范大学 | A kind of Rhodococcus sp DSH of degradable environmental hormone bisphenol-A and its application |
| CN107523513A (en) * | 2017-04-05 | 2017-12-29 | 东北师范大学 | A kind of compound bacteria of 17 β estradiol capable of being fast degraded and its preparation method and application |
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