CN1133742C - 降解卤代烃的微生物及其应用 - Google Patents
降解卤代烃的微生物及其应用 Download PDFInfo
- Publication number
- CN1133742C CN1133742C CNB971907056A CN97190705A CN1133742C CN 1133742 C CN1133742 C CN 1133742C CN B971907056 A CNB971907056 A CN B971907056A CN 97190705 A CN97190705 A CN 97190705A CN 1133742 C CN1133742 C CN 1133742C
- Authority
- CN
- China
- Prior art keywords
- microorganism
- acid
- soil
- halohydrocarbon
- water
- 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.)
- Expired - Fee Related
Links
- 244000005700 microbiome Species 0.000 title claims abstract description 86
- 150000008282 halocarbons Chemical class 0.000 title claims abstract description 13
- XSTXAVWGXDQKEL-UHFFFAOYSA-N Trichloroethylene Chemical group ClC=C(Cl)Cl XSTXAVWGXDQKEL-UHFFFAOYSA-N 0.000 claims abstract description 58
- 239000002689 soil Substances 0.000 claims abstract description 54
- 238000000034 method Methods 0.000 claims abstract description 43
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 30
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 23
- 150000005826 halohydrocarbons Chemical class 0.000 claims description 21
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 15
- 230000015556 catabolic process Effects 0.000 claims description 14
- 238000006731 degradation reaction Methods 0.000 claims description 14
- 241000234282 Allium Species 0.000 claims description 13
- 235000002732 Allium cepa var. cepa Nutrition 0.000 claims description 13
- 239000012190 activator Substances 0.000 claims description 11
- TYFQFVWCELRYAO-UHFFFAOYSA-N suberic acid Chemical compound OC(=O)CCCCCCC(O)=O TYFQFVWCELRYAO-UHFFFAOYSA-N 0.000 claims description 10
- 239000002253 acid Substances 0.000 claims description 9
- FJKROLUGYXJWQN-UHFFFAOYSA-N 4-hydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 claims description 7
- 238000000746 purification Methods 0.000 claims description 7
- OKVJCVWFVRATSG-UHFFFAOYSA-N 3-hydroxybenzyl alcohol Chemical compound OCC1=CC=CC(O)=C1 OKVJCVWFVRATSG-UHFFFAOYSA-N 0.000 claims description 6
- RWZYAGGXGHYGMB-UHFFFAOYSA-N anthranilic acid Chemical compound NC1=CC=CC=C1C(O)=O RWZYAGGXGHYGMB-UHFFFAOYSA-N 0.000 claims description 6
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 claims description 6
- -1 phenyl aldehyde Chemical class 0.000 claims description 6
- YNQLUTRBYVCPMQ-UHFFFAOYSA-N Ethylbenzene Chemical compound CCC1=CC=CC=C1 YNQLUTRBYVCPMQ-UHFFFAOYSA-N 0.000 claims description 5
- 150000001896 cresols Chemical class 0.000 claims description 5
- XCIXKGXIYUWCLL-UHFFFAOYSA-N cyclopentanol Chemical compound OC1CCCC1 XCIXKGXIYUWCLL-UHFFFAOYSA-N 0.000 claims description 5
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 claims description 4
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 claims description 4
- BZHJMEDXRYGGRV-UHFFFAOYSA-N Vinyl chloride Chemical compound ClC=C BZHJMEDXRYGGRV-UHFFFAOYSA-N 0.000 claims description 4
- HPXRVTGHNJAIIH-UHFFFAOYSA-N cyclohexanol Chemical compound OC1CCCCC1 HPXRVTGHNJAIIH-UHFFFAOYSA-N 0.000 claims description 4
- 229940090248 4-hydroxybenzoic acid Drugs 0.000 claims description 3
- LHGVFZTZFXWLCP-UHFFFAOYSA-N guaiacol Chemical compound COC1=CC=CC=C1O LHGVFZTZFXWLCP-UHFFFAOYSA-N 0.000 claims description 3
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 claims 4
- QIRNGVVZBINFMX-UHFFFAOYSA-N 2-allylphenol Chemical compound OC1=CC=CC=C1CC=C QIRNGVVZBINFMX-UHFFFAOYSA-N 0.000 claims 2
- HCPOCMMGKBZWSJ-UHFFFAOYSA-N ethyl 3-hydrazinyl-3-oxopropanoate Chemical compound CCOC(=O)CC(=O)NN HCPOCMMGKBZWSJ-UHFFFAOYSA-N 0.000 claims 2
- 239000001530 fumaric acid Substances 0.000 claims 2
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 claims 2
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 claims 2
- CQRYARSYNCAZFO-UHFFFAOYSA-N salicyl alcohol Chemical compound OCC1=CC=CC=C1O CQRYARSYNCAZFO-UHFFFAOYSA-N 0.000 claims 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 claims 2
- 230000008569 process Effects 0.000 abstract description 5
- 241001453380 Burkholderia Species 0.000 abstract description 4
- UBOXGVDOUJQMTN-UHFFFAOYSA-N trichloroethylene Natural products ClCC(Cl)Cl UBOXGVDOUJQMTN-UHFFFAOYSA-N 0.000 abstract 2
- 210000005239 tubule Anatomy 0.000 description 29
- 235000015097 nutrients Nutrition 0.000 description 28
- 239000002609 medium Substances 0.000 description 18
- 239000007788 liquid Substances 0.000 description 17
- 230000001580 bacterial effect Effects 0.000 description 15
- 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 13
- 229940041514 candida albicans extract Drugs 0.000 description 13
- 239000008103 glucose Substances 0.000 description 13
- 239000012138 yeast extract Substances 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 11
- 239000012071 phase Substances 0.000 description 11
- 239000007789 gas Substances 0.000 description 10
- 239000004809 Teflon Substances 0.000 description 7
- 229920006362 Teflon® Polymers 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 238000011081 inoculation Methods 0.000 description 6
- 235000013343 vitamin Nutrition 0.000 description 6
- 239000011782 vitamin Substances 0.000 description 6
- 229940088594 vitamin Drugs 0.000 description 6
- 229930003231 vitamin Natural products 0.000 description 6
- 150000003722 vitamin derivatives Chemical class 0.000 description 6
- 239000004411 aluminium Substances 0.000 description 5
- 229910052782 aluminium Inorganic materials 0.000 description 5
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 5
- 230000000593 degrading effect Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 239000002351 wastewater Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000589513 Burkholderia cepacia Species 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 241000252867 Cupriavidus metallidurans Species 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 4
- 210000003495 flagella Anatomy 0.000 description 4
- 230000007306 turnover Effects 0.000 description 4
- 241000589776 Pseudomonas putida Species 0.000 description 3
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000005202 decontamination Methods 0.000 description 3
- 230000006866 deterioration Effects 0.000 description 3
- 238000001784 detoxification Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 230000002906 microbiologic effect Effects 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 229910052710 silicon Inorganic materials 0.000 description 3
- 239000010703 silicon Substances 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- 239000011573 trace mineral Substances 0.000 description 3
- 235000013619 trace mineral Nutrition 0.000 description 3
- YQUVCSBJEUQKSH-UHFFFAOYSA-N 3,4-dihydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C(O)=C1 YQUVCSBJEUQKSH-UHFFFAOYSA-N 0.000 description 2
- ALYNCZNDIQEVRV-UHFFFAOYSA-N 4-aminobenzoic acid Chemical compound NC1=CC=C(C(O)=O)C=C1 ALYNCZNDIQEVRV-UHFFFAOYSA-N 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- 241000607142 Salmonella Species 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- 230000003588 decontaminative effect Effects 0.000 description 2
- 230000003413 degradative effect Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 210000003953 foreskin Anatomy 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 238000000053 physical method Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000029058 respiratory gaseous exchange Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- LMWMTSCFTPQVCJ-UHFFFAOYSA-N 2-methylphenol;phenol Chemical compound OC1=CC=CC=C1.CC1=CC=CC=C1O LMWMTSCFTPQVCJ-UHFFFAOYSA-N 0.000 description 1
- JOOXCMJARBKPKM-UHFFFAOYSA-N 4-oxopentanoic acid Chemical compound CC(=O)CCC(O)=O JOOXCMJARBKPKM-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 241000370738 Chlorion Species 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- WAEMQWOKJMHJLA-UHFFFAOYSA-N Manganese(2+) Chemical compound [Mn+2] WAEMQWOKJMHJLA-UHFFFAOYSA-N 0.000 description 1
- 241001575980 Mendoza Species 0.000 description 1
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000589755 Pseudomonas mendocina Species 0.000 description 1
- 241001041887 Pseudomonas putida F1 Species 0.000 description 1
- 241000190932 Rhodopseudomonas Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 150000001491 aromatic compounds Chemical class 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 150000003935 benzaldehydes Chemical class 0.000 description 1
- 235000019445 benzyl alcohol Nutrition 0.000 description 1
- 150000003938 benzyl alcohols Chemical class 0.000 description 1
- 125000005340 bisphosphate group Chemical group 0.000 description 1
- OWBTYPJTUOEWEK-UHFFFAOYSA-N butane-2,3-diol Chemical compound CC(O)C(C)O OWBTYPJTUOEWEK-UHFFFAOYSA-N 0.000 description 1
- 235000004883 caffeic acid Nutrition 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 150000001735 carboxylic acids Chemical class 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 230000009087 cell motility Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- HRYZWHHZPQKTII-UHFFFAOYSA-N chloroethane Chemical compound CCCl HRYZWHHZPQKTII-UHFFFAOYSA-N 0.000 description 1
- 229910001429 cobalt ion Inorganic materials 0.000 description 1
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000005108 dry cleaning Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229910001448 ferrous ion Inorganic materials 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000011087 fumaric acid Nutrition 0.000 description 1
- 150000002238 fumaric acids Chemical class 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- JYVHOGDBFNJNMR-UHFFFAOYSA-N hexane;hydrate Chemical compound O.CCCCCC JYVHOGDBFNJNMR-UHFFFAOYSA-N 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 150000002500 ions Chemical group 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 150000002689 maleic acids Chemical class 0.000 description 1
- 229910001437 manganese ion Inorganic materials 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910001453 nickel ion Inorganic materials 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- FITXJLJVGCNARJ-UHFFFAOYSA-N phenol toluene Chemical compound C1(=CC=CC=C1)C.C1(=CC=CC=C1)O.C1(=CC=CC=C1)C.C1(=CC=CC=C1)O FITXJLJVGCNARJ-UHFFFAOYSA-N 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000003802 soil pollutant Substances 0.000 description 1
- 238000003900 soil pollution Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 150000003442 suberic acids Chemical class 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical class OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- MYVIATVLJGTBFV-UHFFFAOYSA-M thiamine(1+) chloride Chemical compound [Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N MYVIATVLJGTBFV-UHFFFAOYSA-M 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000004506 ultrasonic cleaning Methods 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 229960003487 xylose Drugs 0.000 description 1
Images
Classifications
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
-
- 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
-
- A—HUMAN NECESSITIES
- A63—SPORTS; GAMES; AMUSEMENTS
- A63D—BOWLING GAMES, e.g. SKITTLES, BOCCE OR BOWLS; INSTALLATIONS THEREFOR; BAGATELLE OR SIMILAR GAMES; BILLIARDS
- A63D3/00—Table bowling games; Miniature bowling-alleys; Bowling games
-
- 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
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
-
- 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
-
- 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
- C12R2001/38—Pseudomonas
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/8215—Microorganisms
- Y10S435/822—Microorganisms using bacteria or actinomycetales
- Y10S435/874—Pseudomonas
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Environmental & Geological Engineering (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Water Supply & Treatment (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Molecular Biology (AREA)
- Soil Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Processing Of Solid Wastes (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
属于伯克霍尔德氏菌属并具有降解卤代烃能力的微生物,它们能在2天内将100ppm三氯乙烯降解50%或更多,或在18小时内将30ppm三氯乙烯100%降解,本发明的微生物也提供了一种利用这些微生物降解水或土壤中的卤代烃的方法。
Description
技术领域
本发明涉及被卤代烃污染的水或土壤的生物净化方法。
背景技术
最近,有机溶剂,尤其是卤代烃,在尖端工业中被大量用作清洁剂。由于人们越来越关注由这些物质或含这些物质的废水引起的地下水和土壤污染,有必要立即采取措施对付这种污染。
过去已作为对策使用的已知物理方法的实例包括,空气脱污法(airstripping method),该方法的过程为翻挖土壤,使空气吹过土壤表面,以使卤代烃挥发,并用活性炭等吸附卤代烃;真空抽提法,该方法的过程为将污染的土壤灌入管子中,随后抽真空给土壤换气从而除去污染物。这些方法被认为也可用于地下水的脱污处理。
然而,这些方法的缺点是需要大量能量,如要鼓风。而且,前一种方法还有一个缺点是需要翻挖土壤,而后一种方法的另一个缺点是提取效率随着污染物浓度下降而降低,从而使得净化难以实现。另外,从防止空气污染等二次污染的观点看,为了将分离的污染物吸附至活性炭等上面,这些方法要求对它们进行脱毒。
另一方面,近年已进行了对所谓生物净化方法的研究,生物净化方法就是利用微生物使污染物有效地降解和脱毒。由于这些方法利用微生物的降解机制,与上述物理方法相比,它们无需大量的能量。它们也能够完全降解污染物和使之脱毒而不会引起二次污染。而且,即使污染物处于低浓度也可进行净化,从而使得脱污可以在原始发生地的广泛区域进行,并极有希望降低费用。
采用微生物净化污染土壤的方法的实例包括,固相处理法,该方法是将微生物与磷、氮等营养源混入翻挖的土壤中促进污染物的降解;泥浆处理法,该方法是将微生物与水和营养源混入翻挖的土壤中,以流体形式处理土壤,促进污染物的降解;原地处理法,该方法是将空气、营养源等注入污染土壤中,无需翻挖而促进土壤中存在的微生物降解污染物。
在上述生物处理方法中,固相处理法和泥浆处理法由于需要翻挖土壤,应用范围受到限制,处理和设备费用相对较高。
另一方面,原地处理法费用相对较低,并且可以对广泛区域进行处理。然而,其净化速率较慢,因为土壤微生物的绝对数量较少。在待处理化合物难以降解,尤其如为卤代烃时,土壤中有可能不存在能降解土壤污染物的微生物。在这种情况下,获取能够降解卤代烃的微生物并将它们接种至土壤中,使得净化速率提高,并且即使土壤中不存在能够降解污染物的微生物,土壤也可得以净化。
一种卤代烃污染物三氯乙烯(TCE)广泛用于IC工业、干洗等当中。它是一种特别重要的污染物,因为据报道它可致癌。降解TCE的微生物的已知实例包括甲烷同化微生物Methyrosinus tricosporium OB3(日本未审查专利出版号4-501667,日本未审查专利出版号5-212371)和Methyrosinus tricosporium TUKUBA(日本未审查专利出版号2-92274和日本未审查专利出版号3-292970),
假单胞菌,如恶臭假单胞菌F1(日本未审查专利出版号64-34499);恶臭假单胞菌BH(Fujita等,“化学工程”39卷6期494-498页,1994)、恶臭假单胞菌UC-R5和UC-P2(日本未审查专利出版号62 84780)、恶臭假单胞菌KWI-9(日本未审查专利出版号6-70753)、门多萨假单胞菌KR-1(日本未审查专利出版号2-503866和5-502593)、洋葱假单胞菌G4(日本未审查专利出版号4-502277)和洋葱假单胞菌KK01(日本未审查专利出版号6-296711)以及其它微生物,如真养产碱菌JMP134(A.R.Harker,“应用与环境微生物学”56卷4期1179-1181页,1990)、真养产碱菌KS01(日本未审查专利出版号7-123976)和氨细菌Nitrosomonuseuropaea(D.Arciero等,“生物化学与生物物理研究通讯”159卷2期640-643页,1989)是已知的。
特别是洋葱假单胞菌KK01据报道可降解液体培养物中初始浓度为30pprn的TCE到15ppm,土壤中初始浓度为5ppm的TCE到1ppm(日本未审查专利出版号6-296711)。另外据报道真养产碱菌可降解液体培养物中浓度为50ppm的TCE至低于检测水平,土壤中浓度为低于检测水平的1ppm的TCE(日本未审查专利出版号7-123976)。
然而,测定这些微生物的降解能力时,降解均在极高细胞浓度(1×108细胞/毫升)下得以证实。考虑到该浓度在实际土壤环境中根本不存在,不能认为在各种情况下这些微生物的降解能力都强。因而使用微生物净化土壤时,这些微生物应具有充分的降解能力,并且能在特殊的土壤环境,如存在野生微生物时,表现该能力。另外,优选这些微生物对降解目标物TCE的耐受性高,并且它们也能降解二氯乙烯(DCE),它是TCE的部分降解物。
发明内容
本发明的目的涉及有效降解卤代烃,尤其是高浓度TCE和DCE等的微生物,以及使用这些微生物对水或土壤进行净化的方法。
本发明提供能降解卤代烃、属于伯克霍尔德氏菌属的微生物,一些这样的微生物属于洋葱伯克霍尔德氏菌。这些微生物的实例包括伯克霍尔德氏菌N16-1(FERM BP-5504)、洋葱伯克霍尔德氏菌N15-1(FERM BP-5502)和洋葱伯克霍尔德氏菌N15-2(FERM BP-5503)。而且,本发明提供将上述微生物加入含卤代烃的水或土壤来净化水或土壤的方法。
本发明还有另一个特点就是将微生物激活剂与上述微生物一起加入。这些微生物能在2天内将100ppm三氯乙烯降解50%或更多,或在18小时内将30ppm三氯乙烯完全降解。
附图简述
图1示出实施例2中残留TCE的百分率随时间的变化。
图2示出实施例3中残留TCE的百分率随时间的变化。
图3示出实施例4中残留顺式-1,2-DCE的百分率随时间的变化。
图4示出实施例6中残留TCE的百分率。
图5示出实施例7中残留TCE的百分率。
详述
本发明的微生物应为属于伯克霍尔德氏菌属或属于洋葱伯克霍尔德氏菌的微生物,这些微生物的具体实例包括伯克霍尔德氏菌N16-1、洋葱伯克霍尔德氏菌N15-1和洋葱伯克霍尔德氏菌N15-2。这些微生物为由自然界(如河流和土壤)中新分离的菌株,它们的分离方法和分类特征将具体描述。这些菌株,即伯克霍尔德氏菌N16-1,洋葱伯克霍尔德氏菌N15-1和洋葱伯克霍尔德氏菌N15-2,于1996年4月12日保藏在国立生命科学和人体技术研究所,保藏号分别为FERM BP-5504、FERM BP-5502和FERM BP-5503。
本发明的微生物可在含常用碳源、氮源以及必需的无机盐、维生素和其它痕量元素的培养基中培养。本发明的微生物优先同化的任何碳源均可使用。培养基中碳源浓度优选例如0.1到0.5g/L,但应根据碳源类型而有所变化。可以使用的氮源的实例包括有机氮源,如酵母提取物、蛋白胨和肉提取物,而无机氮源的实例包括铵盐和硝酸盐。
氮源浓度优选0.1到1.4g/L,但应根据具体类型而有所变化。无机盐的优选实例包括金属离子,如钾离子、钙离子、镁离子、二价铁离子、锰离子、钴离子和镍离子,以及阴离子,如氯离子、硫酸根离子和磷酸根离子。优选通气培养,振荡培养或大规模培养时优选通气并搅拌。培养温度为20到37℃,优选大约30℃。
另外,本发明涉及一种净化水或土壤的方法,其特征在于将上述微生物加入含卤代烃的水或土壤中。在该方法中,如上述方式培养的本发明的微生物应加入待处理的水或土壤中。这些微生物应以培养液或自培养液中分离的微生物的形式加入。另外,这些微生物也可吸附到分开的载体上之后再加入。
尽管加入的微生物的量根据微生物降解卤代烃的能力、待处理的水或土壤中卤代烃的量等而有所变化,但该量在105到109细胞/克之间。尽管处理所需的时间也根据使用的微生物降解卤代烃的能力、待处理的水或土壤中卤代烃的量和加入的微生物的量而有所不同,但它大约为1到10天。
另外本发明对水或土壤的净化方法涉及一种净化水或土壤的方法,该方法是将如上述的本发明的微生物接种被卤代烃污染的水或土壤并混合,以降解水或土壤中所含的卤代烃,并且上述对水或土壤的净化方法,其特征在于当在上述水或土壤中接种并混合上述微生物时,加入并混合至少一类微生物激活剂。在这种情况下,微生物激活剂具有激活微生物降解卤代烃能力的作用,并被称为诱导剂。可以使用的诱导剂的实例包括能被上述微生物同化和降解的化合物,优选的实例包括苯、甲苯、酚、甲酚和3-羟基苄醇。另外,除上述诱导剂外,N16-1还可使用环戊醇、己酸、反式-3-己酸和辛二酸。
可被本发明的方法降解的卤代烃的实例尤为氯代烃,如TCE、DCE和一氯乙烯。
尽管本发明的方法可用于前述的固相处理法和泥浆处理法,但也可不使用这些需要翻挖土壤的方法,而简单地通过加入和接种本发明的微生物至土壤或水中而进行净化。
实施例
下面通过实施例详细说明本发明。实施例1.微生物的分离与鉴定
本发明的微生物使用下述方法从一家化工厂厂区的土壤中筛选和分离。0.1g土壤样品接种至25ml螺口试管中的5ml NMS培养基或M9培养基。并且,加入500ppm酚和维生素混合液,随后盖上试管盖,按规定时间于30℃振荡培养。培养至培养基由于其中微生物的生长而变浑浊为止。传代培养10次后,将培养液适当稀释,用棉拭子接种加入1.5%琼脂的平板培养基以分离长出的微生物菌落。重复该步骤分离微生物。
表1
NMS培养基的成分(1升中)
MgSO4·7H2O 1.0g
CaCl2 0.01g
Na2HPO4·12H2O 0.717g
NH4Cl 0.6g
KH2PO4 0.272g痕量元素溶液(pH6.8) 0.5ml
痕量元素溶液(1升中)
EDTA 500mg
FeSO4·7H2O 200mg
ZnSO4·7H2O 10mg
MnCl2·4H2O 3mg
H3BO3 30mg
CoCl2·6H2O 20mg
NiCl2·6H2O 2mg
CaCl2 1mg
Na2MoO4·2H2O 2mg
维生素混合液(1升中)
盐酸硫胺素 3mg
对氨基苯甲酸 13mg
腺嘌呤 1.0g
NAD 0.25g
维生素B12 10mg
二磷酸氯化硫胺素 100mg
分离的微生物在液体培养基中培养2天以后,1/100体积的培养液接种至含4ml NMS培养基的30ml小管中,培养基中加入有酚、0.02%的酵母提取物和1mM葡萄糖,随后力入10ppm TCE。用覆以特氟隆的隔膜盖和铝盖迅速密封小管,30℃振荡培养5天,用气相色谱分析小管中的气相。对这样选出的3株具有高水平TCE降解活性的微生物的形态和生理特性进行了研究。结果于表2中示出。
表2
试验参数 试验结果
N15-1 N15-2 N16-1形态 杆状 杆状 杆状革兰氏染色 - - -芽胞 - - -运动性 + + +鞭毛 极生多鞭毛 极生多鞭毛 极生多鞭毛对氧气反应 需氧 需氧 需氧氧化酶 + + +触酶 + + +OF O O O菌落颜色色泽 NP NP NP荧光色素形成 - - -水溶性色素形成 - - -PHB积累 + + +裂解原儿茶酸 邻位型 邻位型 邻位型精氨酸双水解酶 - - -40℃生长 + + -反硝化作用 - - -硝酸盐还原 + + -明胶液化 + + -淀粉水解 - - -同化葡萄糖 + + +木糖 + + +鼠李糖 - - +乙酰丙酸 4 + -甲基延胡索酸 - - -D-酒石酸 - - +2,3-丁二醇 + + -色胺 - - -醌型 Q-8 Q-8 Q-8胞内DNA GC含量 66 67 62(ml%)
根据文献(N.R.Kreig和J.G.Holt,《伯杰氏系统细菌学手册》第1卷(1984)Williams&Wilkins;J.G.Holt,N.R.Krieg,P.H.A.Sneath,J.T.Staley和ST.Williams《伯杰氏细菌学鉴定手册》第9版(1994)Williams&Wilkins;N.Zhao,C.Qu,E.Wang和W.Chen,“国际系统细菌学杂志”45卷600页(1995);E.Yabuuti,Y.Kosako,H Oyaizu,I.Yano,H.Hotta,Y.Hashimoto,T.Ezaki和M.Arakawa,“微生物学与免疫学”36卷1251页(1992)),利用上述结果对上述微生物进行了鉴定,一个菌株被鉴定为伯克霍尔德氏菌属的种类,而另两个菌株被鉴定为洋葱伯克霍尔德氏菌,并分别命名为N16-1、N15-1和N15-2。
如前所述,属于洋葱伯克霍尔德氏菌、降解TCE的已知微生物为G4(日本未审查专利出版号4-502277)和KK01(日本未审查专利出版号6-296711)。由表4可见N15-1和N15-2均具运动性,因而它们明显区别于不具运动性的G4。另外,G4和KK01被甲苯诱导,N15-1和N15-2不被甲苯诱导,因而它们也可据此参数明显地区分。另一方面,由于N16-1胞内DNA的GC含量为62mol%,与已知降解TCE的微生物恶臭伯克霍尔德氏菌、门多萨伯克霍尔德氏菌或洋葱伯克霍尔德氏菌(1992年以前被划入假单胞菌属)不同,它被鉴定为一种新的微生物。实施例2.在液体培养基中降解TCE
菌株N15-1、N15-2和N16-1均在加有500ppm酚和维生素混合液的NMS液体培养基中培养一天。离心收集微生物,随后重悬于4ml不含酚的同一培养基中。将30ml该悬液转入小管中,随后加入100ppm TCE,小管用覆以特氟隆的硅隔膜和铝盖迅速密封(微生物浓度108细胞/ml)。小管随后静置于30℃培养。按时以气相色谱分析气相。这些结果于图1中示出。每种微生物在5天内降解了液体培养基中超过70%的TCE。实施例3.在液体培养基中降解TCE
菌株N15-1、N15-2和N16-1均在加有0.2%酵母提取物和4mM葡萄糖的NMS液体培养基中培养1天。在小管中加入4ml加有0.02%酵母提取物、500ppm酚和1mM葡萄糖的NMS液体培养基,取上述培养液(微生物计数大约106细胞/毫升)各40μl接种。加入100ppm TCE,随后用覆以特氟隆的硅隔膜和铝盖迅速密封小管。小管于30℃振荡培养,以气相色谱按时对气相进行分析。结果于图2中示出。
每种微生物在10天内降解了培养液中超过60%的TCE。当将该降解能力与已知TCE降解微生物相比较时,只有两篇报道提到超过30ppm的TCE。其中,洋葱假单胞菌KK01(日本未审查专利出版号6-296711)据报道可在2天内降解30ppm TCE下降至大约15ppm(50%),而真养产碱菌KS01据报道在细胞浓度为108细胞/毫升时,能在4天内降解50ppm TCE至低于气相色谱检测水平。相形之下,由于N15-1、N15-2和N16-1能降解浓度高达100ppm的TCE,而且每一细胞的降解能力也较高,因而除了在实际应用时能够适应较宽的污染浓度范围之外,还具有能够降低费用的优点,因为所需的微生物量大大减少了。实施例4.在液体培养基中降解DCE
菌株N15-1、N15-2和N16-1均接种5ml补加有0.02%酵母提取物、500ppm酚和1mM葡萄糖的NMS液体培养基。30℃培养2天后,1/100体积的培养液被加入含4ml加有30ppm顺式-1,2-DCE的同样培养液的小管中,30℃培养5天。结果于图3中示出。三种菌株均可见降解超过99%的DCE。实施例5.降解期间温度的影响
菌株N15-1、N15-2和N16-1均接种5ml补加有0.02%酵母提取物、500ppm酚和1mM葡萄糖的NMS液体培养基中。30℃培养2天后,1/100体积的培养液被加入含4ml加有30ppm TCE的同样培养液的小管中,随后在16到30℃培养8天。对N16-1和N15-1来说,在各种温度TCE均被降解至低于检测水平。而对N15-2来说,尽管16℃时残留有大约30%的TCE,但在20℃或更高温度时,残余量低于检测水平。因而,这表明这些微生物能在土壤温度(15到20℃)下降解TCE。实施例6.降解期间pH的影响
菌株N15-1、N15-2和N16-1均接种5ml补加有0.02%酵母提取物、500ppm酚和1mM葡萄糖的M9液体培养基(pH7.0)。30℃培养2天后,1/100体积的培养液被加入含4ml培养基的小管中,培养基pH被调整为5-10,其中补加有30ppm TCE而不是酚,随后于30℃培养5天。N15-1、N15-2和N16-1在各种pH时均降解30ppm TCE至低于检测水平。另外,尽管N16-1的生长在pH7.4或更高时受到抑制,但它在pH5到7之间降解TCE至低于检测水平。实施例7.土壤中TCE降解试验
10g Andsols(样品取自Aichi prefecture并风干)加入体积为30ml的小管中,随后加入TCE至浓度为20ppm。菌株N15-1、N15-2和N16-1均接种20ml补加有0.02%酵母提取物、500ppm酚和1mM葡萄糖的NMS液体培养基。30℃振荡培养3天,通过离心由培养液中收集微生物细胞,细胞重悬于适当体积、不含酚的NMS培养基中,上述小管接种细胞的量为108到109细胞/克,加入悬液后水分含量为25%。用外罩覆以特氟隆包皮的螺旋盖盖住小管,振荡,小管于30℃温育7天。
称取10g土壤,置于带塞子的锥形瓶中,随后加入90ml以通过活性炭的空气充气的离子交换水、5ml磷酸和10ml正己烷。将烧瓶密闭后,在超声净化器中超声处理20分钟,随后于摇床上摇5分钟。之后,将水相和正己烷相转移至有盖比色杯中。将比色杯密封,进行超声处理,分离的正己烷相通过气相色谱分析。结果示于图4中。
初始浓度为10ppm的TCE在7天内降解了30%到50%。因而,本发明的微生物即使在天然环境中也具有降解能力。所以,通过将土壤与本发明的微生物和水接触有可能使污染的土壤净化。并且,该方法无需直接将酚等激活剂加入土壤中而使土壤净化。实施例8.土壤中的TCE降解试验
10g Andsols(样品取自Aichi prefecture并风干)置于体积为30ml的小管中,随后加入TCE至浓度为20ppm。菌株N15-1、N15-2和N16-1均接种20ml NMS液体培养基,培养基中补加有0.02%的酵母提取物、500ppm酚和1mM葡萄糖。随后于30℃振荡培养1天。
取出0.2ml该培养液,加入上述小管中,使得接种微生物的细胞量为106细胞/克,加入培养基后水分含量为40%,酚浓度为500ppm。用外罩覆以特氟隆包皮的螺旋盖盖住小管并振荡,培养液于30℃培养3个星期。结果示于图5中。40%到50%的TCE被降解,说明本发明的微生物即使在106细胞/克的低浓度下也能降解取自天然环境的土壤中的TCE。实施例9.激活剂对降解活性的诱导
菌株N15-1、N15-2和N16-1均于30℃在5ml NMS液体培养基中培养2天,培养基中补加有0.2%酵母提取物和5mM葡萄糖。向小管中加入4ml NMS液体培养基,培养基中补加有0.02%酵母提取物、1mM葡萄糖和100ppm表4中所列的一种激活剂,随后以40μl上述每一种培养液(微生物计数为约106细胞/克)接种。向每一小管中加入30ppmTCE,随后用覆以特氟隆的硅隔膜和铝盖迅速密封小管。小管于30℃振荡培养5天,气相以气相色谱分析。结果示于表5中。表中数字表示加入微生物期间残余TCE浓度的百分比,未加入微生物时的TCE浓度为100%。结果示于以下表3中。
表3
化合物 微生物菌株
N16-1 N15-1 N15-2环己醇 8.6 57.4 64.1环戊醇 49.3 100 100邻氨基苯甲酸 13.3 105 107咖啡酸 52.6 96.7 102辛二酸 7.3 99.8 100马来酸 53.0 100 98.8延胡索酸 63.4 100 100琥珀酸 63.3 72.4 92.4丙二酸 45.2 100 100反-3-已酸 2.7 100 100己酸 8.3 100 100苯 0 98.8 97.3乙苯 28.7 98.4 101苄醇 0 71.9 72.5水杨苷 0 102 105烯丙基酚 0.5 100 102邻甲氧基苯酚 35.0 99.2 107甲苯 0 100 100苯甲醛 6.8 103 99.0对羟基装甲酸 12.5 102 98.8
N15-1和N15-2被酚、甲苯和苯等芳香族化合物激活,这些化合物是已知的常规激活诱导剂,N15-1和N15-2也被非芳香族化合物环己醇激活。另外,琥珀酸也表现出激活这些微生物的能力。并且,N16-1也被环戊醇、邻氨基苯甲酸、对羟基苯甲酸、辛二酸、反式-3-己酸以及己酸等直链羧酸激活。
并且,本发明的菌株N15-1、N15-2和N16-1与已知的洋葱伯克霍尔德氏菌KK01和G4的特性比较示于以下表4中。
表4
特性 洋葱伯克霍尔德氏菌 伯克霍尔德氏菌属
KK01 G4 N15-1,2 N16-1降解能力 30->15/2天, 1->0.17/1天 100->40/2天 100->45/天,
机体数目未知 3×108细胞 108细胞 108细胞/毫升
/毫升 /毫升最大TCE浓度 30 2 100 100(ppm)激活剂 甲苯、酚、 甲苯、酚 甲苯不激活 甲苯、甲酚
甲酚 甲酚 酚、甲酚、环己 环己醇、辛二酸、
醇 其它运动性 + - + +
根据本发明,本发明的微生物能降解水或土壤中所含的高浓度卤代烃,如TCE和DCE,优选在至少一类激活剂和糖或其它营养物质存在下进行降解。
根据本发明中净化水或土壤的方法,除了具有生物技术的优势,如不需大量能量和防止产生二次污染之外,本发明的方法还能够在天然环境中工业化地有效净化卤代烃污染的水或土壤。实施例10.在液体培养中降解三氯乙烯
菌株N15-1、N15-2和N16-1分别在补加有0.2%酵母提取物和5mM葡萄糖的NMS液体培养基中培养1天。每一小管中加入4ml补加有0.02%酵母提取物、100ppm酚和1mM葡萄糖的NMS液体培养基,如上准备的40μl培养物接种小管(大约106细胞/毫升培养基)。向小管中加入三氯乙烯使其浓度为30ppm,小管以覆以特氟隆的硅基子和铝盖封闭。小管于30℃振荡温育,小管中气相定期以气相色谱进行分析。结果,在18小时内三氯乙烯100%被降解。
Claims (10)
1.属于伯克霍尔德氏菌属、具有降解卤代烃能力的微生物,所述卤代烃选自三氯乙烯、二氯乙烯和一氯乙烯,它们能在2天内将100ppm三氯乙烯降解50%或更多,或在18小时内将30ppm三氯乙烯100%降解。
2.根据权利要求1的微生物,其中卤代烃为三氯乙烯。
3.权利要求1或2的微生物,其为洋葱伯克霍尔德氏菌。
4.根据权利要求1或2的微生物,其为伯克霍尔德氏菌N16-1,FERM BP-5504。
5.根据权利要求3的微生物,其为洋葱伯克霍尔德氏菌N15-1,FERM BP-5502。
6.根据权利要求3的微生物,其为洋葱伯克霍尔德氏菌N15-2,FERM BP-5503。
7.一种净化含卤代烃的水或土壤的方法,包括将根据权利要求1的微生物之一或全体加入含卤代烃的水或土壤的步骤,其中所述卤代烃选自三氯乙烯、二氯乙烯和一氯乙烯。
8.根据权利要求7的方法,其中在加入微生物的同时将微生物激活剂加入有关的水或土壤中,其中所述微生物激活剂选自苯酚、甲酚、3-羟基苄醇、环己醇、环戊醇、邻氨基苯甲酸、咖啡酸、辛二酸、马来酸、延胡索酸、琥珀酸、丙二酸、反式-3-己酸、己酸、苯、乙苯、苄醇、水杨苷、烯丙基酚、邻甲氧基苯酚、甲苯、苯甲醛和对羟基苯甲酸。
9.一种净化含卤代烃的水或土壤的方法,包括将根据权利要求2的微生物之一或全体加入含卤代烃的水或土壤的步骤,其中所述卤代烃选自三氯乙烯、二氯乙烯和一氯乙烯。
10.根据权利要求9的方法,其中在加入微生物的同时将微生物激活剂加入有关的水或土壤中,其中所述微生物激活剂选自苯酚、甲酚、3-羟基苄醇、环己醇、环戊醇、邻氨基苯甲酸、咖啡酸、辛二酸、马来酸、延胡索酸、琥珀酸、丙二酸、反式-3-己酸、己酸、苯、乙苯、苄醇、水杨苷、烯丙基酚、邻甲氧基苯酚、甲苯、苯甲醛和对羟基苯甲酸。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP100466/1996 | 1996-04-22 | ||
JP100466/96 | 1996-04-22 | ||
JP10046696 | 1996-04-22 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1195373A CN1195373A (zh) | 1998-10-07 |
CN1133742C true CN1133742C (zh) | 2004-01-07 |
Family
ID=14274694
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNB971907056A Expired - Fee Related CN1133742C (zh) | 1996-04-22 | 1997-04-18 | 降解卤代烃的微生物及其应用 |
Country Status (8)
Country | Link |
---|---|
US (1) | US5998198A (zh) |
EP (1) | EP0840783A1 (zh) |
KR (1) | KR100285035B1 (zh) |
CN (1) | CN1133742C (zh) |
CA (1) | CA2222211C (zh) |
ID (1) | ID17199A (zh) |
MY (1) | MY118003A (zh) |
WO (1) | WO1997040136A1 (zh) |
Families Citing this family (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999044422A1 (fr) * | 1998-03-06 | 1999-09-10 | Toyota Jidosha Kabushiki Kaisha | Procede de controle de micro-organismes cibles |
JP2904432B1 (ja) * | 1998-05-18 | 1999-06-14 | 農林水産省農業環境技術研究所長 | 土壌中の有機塩素系化合物を好気的に分解する分解菌、その分解菌の集積又は単離方法及びその分解菌を保持する分解菌保持担体 |
US20030059866A1 (en) * | 2001-09-26 | 2003-03-27 | Kim Lewis | Isolation and cultivation of microorganisms from natural environments and drug discovery based thereon |
US20050130256A1 (en) * | 2001-09-26 | 2005-06-16 | Northeastern University | Isolation and cultivation of microorganisms from natural environments and drug discovery based thereon |
US7011957B2 (en) * | 2001-09-26 | 2006-03-14 | Northeastern University | Isolation and cultivation of microorganisms from natural environments and drug discovery based thereon |
US7172895B2 (en) * | 2002-04-25 | 2007-02-06 | Takuya Kitamura | Microorganism and drainage method |
CN100391867C (zh) * | 2005-11-04 | 2008-06-04 | 中国地质大学(武汉) | 耐氯菌株3号的应用 |
US7867750B2 (en) * | 2006-07-20 | 2011-01-11 | Rutgers, The State University Of New Jersey | Method for anaerobic degradation of fuel oxygenates and similar compounds |
CN102321659A (zh) * | 2011-08-30 | 2012-01-18 | 浙江工业大学 | 氯代脂肪烃降解性质粒pRC11、工程菌及其应用 |
CN102533593B (zh) * | 2011-12-16 | 2013-06-19 | 华南农业大学 | 洋葱伯克霍尔德氏菌sd7及其培养方法和应用 |
CN102533619B (zh) * | 2012-02-29 | 2013-09-04 | 华东理工大学 | 一种氯代烃高效好氧降解混合菌剂的制备方法与应用 |
CN103451118B (zh) * | 2012-06-04 | 2015-06-03 | 华中农业大学 | 一种戊二醛降解菌株及制备方法和应用 |
CN105194831B (zh) * | 2015-09-30 | 2018-05-18 | 哈尔滨工业大学 | 一种电刺激促进挥发性氯代烃生物还原分解的方法 |
CN105753178B (zh) * | 2016-04-28 | 2019-04-23 | 上海市环境工程设计科学研究院有限公司 | 一种强化微生物原位修复氯代烃污染地下水的方法 |
CN109201727B (zh) * | 2018-09-03 | 2020-12-11 | 杭州鸿明市政工程有限公司 | 一种土壤淋洗液、使用方法和设备 |
CN113684148B (zh) * | 2021-08-19 | 2023-04-14 | 云南中烟工业有限责任公司 | 一株洋葱伯克霍尔德氏菌及其分离培养方法和应用 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1992019738A1 (en) * | 1991-05-02 | 1992-11-12 | Sbp Technologies, Inc. | Microbial degradation of trichloroethylene, dichloroethylenes and aromatic pollutants |
JP3435426B2 (ja) * | 1995-11-30 | 2003-08-11 | トヨタ自動車株式会社 | ハロゲン化炭化水素分解菌及びその使用 |
-
1997
- 1997-04-17 MY MYPI97001671A patent/MY118003A/en unknown
- 1997-04-18 WO PCT/JP1997/001359 patent/WO1997040136A1/en not_active Application Discontinuation
- 1997-04-18 EP EP97917443A patent/EP0840783A1/en not_active Withdrawn
- 1997-04-18 KR KR1019970709474A patent/KR100285035B1/ko not_active IP Right Cessation
- 1997-04-18 US US08/981,352 patent/US5998198A/en not_active Expired - Lifetime
- 1997-04-18 CN CNB971907056A patent/CN1133742C/zh not_active Expired - Fee Related
- 1997-04-18 CA CA002222211A patent/CA2222211C/en not_active Expired - Fee Related
- 1997-04-22 ID IDP971333A patent/ID17199A/id unknown
Non-Patent Citations (1)
Title |
---|
APPLIEDANDENVIRONMENTALMICROBIOLGY,FEB1995 1995-02-01 WINKLER,J,ET,AL,TRACKING,THE,RESPONSE,OF,BURKHOLDERIA,CEPACEA,G4,5223-PR1,IN,Aquifer,MICROBOSMS * |
Also Published As
Publication number | Publication date |
---|---|
KR19990023004A (ko) | 1999-03-25 |
MY118003A (en) | 2004-08-30 |
CA2222211C (en) | 2001-02-13 |
EP0840783A1 (en) | 1998-05-13 |
CA2222211A1 (en) | 1997-10-30 |
ID17199A (id) | 1997-12-11 |
WO1997040136A1 (en) | 1997-10-30 |
KR100285035B1 (ko) | 2001-04-02 |
CN1195373A (zh) | 1998-10-07 |
US5998198A (en) | 1999-12-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1133742C (zh) | 降解卤代烃的微生物及其应用 | |
Mawang et al. | Actinobacteria: An eco-friendly and promising technology for the bioaugmentation of contaminants | |
Ma et al. | Combination of biochar and immobilized bacteria accelerates polyacrylamide biodegradation in soil by both bio-augmentation and bio-stimulation strategies | |
Safonova et al. | Biotreatment of industrial wastewater by selected algal‐bacterial consortia | |
Jun et al. | Isolation and identification of a di-(2-ethylhexyl) phthalate-degrading bacterium and its role in the bioremediation of a contaminated soil | |
Zhenggang et al. | Biosorption characteristics of Mn (II) by Bacillus cereus strain HM-5 isolated from soil contaminated by manganese ore | |
Semenyuk et al. | Effect of activated charcoal on bioremediation of diesel fuel-contaminated soil | |
CA2596386C (en) | Surfactant biocatalyst for remediation of recalcitrant organics and heavy metals | |
Fulekar | Bioremediation Technology for hazardous wastes-recent advances | |
CN101735996A (zh) | 一种修复多环芳烃污染场地的混合菌剂及制备方法和应用方法 | |
CN109354179A (zh) | 一种反硝化生物膜脱氮生物反应器、其使用方法和应用 | |
CN100355875C (zh) | 一种处理炼油废水的微生物菌剂、其制备方法及其应用 | |
EP1210407B1 (en) | Bacterial consortium ebc1000 and a method using the bacterial consortium ebc1000 for remedying biologically recalcitrant toxic chemicals contained in industrial wastewater, waste materials and soils | |
Ossai et al. | Biological treatments for petroleum hydrocarbon pollutions: The eco-friendly technologies | |
Kumar et al. | Biological Approaches to Controlling Pollutants | |
JP6999893B2 (ja) | 石油汚染土壌の浄化用組成物の製造方法 | |
CN109370957B (zh) | 一种污水处理复合菌剂及其制备方法 | |
CN1571833A (zh) | 能够降解甲基叔丁基醚的多噬菌菌株及其用途 | |
Buraimoh et al. | Efficacy of intervention strategies for bioremediation of crude oil in polluted soil microcosm | |
JP4203546B2 (ja) | 分解方法および浄化方法 | |
JP3436654B2 (ja) | ハロゲン化炭化水素分解菌及びその使用 | |
US20090277831A1 (en) | Microbial based chlorinated ethene destruction | |
Lutsinge | Biosurfactant enhanced biodegradation of high molecular weight polycyclic aromatic hydrocarbons in a two-stage continuous stirred tank bioreactors and biofilm tank | |
JP3435426B2 (ja) | ハロゲン化炭化水素分解菌及びその使用 | |
Kazankapova et al. | Treatment of oil-containing wastewater using microorganisms immobilized on shungite |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C19 | Lapse of patent right due to non-payment of the annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |