CN110004136A - A kind of embedding body, preparation method and cultural method loading leaching microbacteria - Google Patents
A kind of embedding body, preparation method and cultural method loading leaching microbacteria Download PDFInfo
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- CN110004136A CN110004136A CN201910302360.XA CN201910302360A CN110004136A CN 110004136 A CN110004136 A CN 110004136A CN 201910302360 A CN201910302360 A CN 201910302360A CN 110004136 A CN110004136 A CN 110004136A
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- China
- Prior art keywords
- leaching
- leaching microbacteria
- embedding
- nutrient
- microbacteria
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- 238000002386 leaching Methods 0.000 title claims abstract description 116
- 238000000034 method Methods 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 238000011068 loading method Methods 0.000 title claims abstract description 10
- 235000015097 nutrients Nutrition 0.000 claims abstract description 72
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 34
- 239000006187 pill Substances 0.000 claims abstract description 27
- 239000000126 substance Substances 0.000 claims abstract description 20
- 239000002245 particle Substances 0.000 claims abstract description 6
- 239000000945 filler Substances 0.000 claims abstract description 4
- 239000011258 core-shell material Substances 0.000 claims abstract description 3
- 239000012528 membrane Substances 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 115
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 claims description 27
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 24
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 18
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 18
- 229910001448 ferrous ion Inorganic materials 0.000 claims description 18
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 17
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 14
- 239000000661 sodium alginate Substances 0.000 claims description 14
- 235000010413 sodium alginate Nutrition 0.000 claims description 14
- 229940005550 sodium alginate Drugs 0.000 claims description 14
- ZCCIPPOKBCJFDN-UHFFFAOYSA-N calcium nitrate Chemical compound [Ca+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ZCCIPPOKBCJFDN-UHFFFAOYSA-N 0.000 claims description 12
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 12
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 12
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 11
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 11
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 11
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 10
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical group O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 9
- 239000007864 aqueous solution Substances 0.000 claims description 8
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims description 7
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 7
- 239000001103 potassium chloride Substances 0.000 claims description 7
- 235000011164 potassium chloride Nutrition 0.000 claims description 7
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims description 6
- UKVHQGHRJIEIML-UHFFFAOYSA-L calcium boric acid dichloride Chemical compound [Cl-].[Cl-].[Ca+2].OB(O)O UKVHQGHRJIEIML-UHFFFAOYSA-L 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 claims description 4
- 238000004132 cross linking Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 229910001414 potassium ion Inorganic materials 0.000 claims description 3
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 claims description 2
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 claims description 2
- 229910001424 calcium ion Inorganic materials 0.000 claims description 2
- 229910001425 magnesium ion Inorganic materials 0.000 claims description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims 1
- 239000011777 magnesium Substances 0.000 claims 1
- 229910052749 magnesium Inorganic materials 0.000 claims 1
- 235000008935 nutritious Nutrition 0.000 claims 1
- 244000005700 microbiome Species 0.000 abstract description 82
- 230000008569 process Effects 0.000 abstract description 9
- 230000006978 adaptation Effects 0.000 abstract description 2
- 231100000572 poisoning Toxicity 0.000 abstract description 2
- 230000000607 poisoning effect Effects 0.000 abstract description 2
- 235000016709 nutrition Nutrition 0.000 description 40
- 230000035764 nutrition Effects 0.000 description 35
- 239000008188 pellet Substances 0.000 description 28
- 230000000694 effects Effects 0.000 description 27
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 26
- 241000894006 Bacteria Species 0.000 description 21
- 230000012010 growth Effects 0.000 description 19
- 239000002609 medium Substances 0.000 description 18
- 239000002028 Biomass Substances 0.000 description 16
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 16
- 229910052698 phosphorus Inorganic materials 0.000 description 16
- 239000011574 phosphorus Substances 0.000 description 16
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 15
- 229940068984 polyvinyl alcohol Drugs 0.000 description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 15
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 14
- 239000011790 ferrous sulphate Substances 0.000 description 14
- 235000003891 ferrous sulphate Nutrition 0.000 description 14
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 14
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 14
- 229910052742 iron Inorganic materials 0.000 description 13
- 230000000813 microbial effect Effects 0.000 description 13
- 239000012153 distilled water Substances 0.000 description 12
- 239000000463 material Substances 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 11
- 229910001447 ferric ion Inorganic materials 0.000 description 11
- 241000605222 Acidithiobacillus ferrooxidans Species 0.000 description 10
- 229910000628 Ferrovanadium Inorganic materials 0.000 description 10
- PNXOJQQRXBVKEX-UHFFFAOYSA-N iron vanadium Chemical compound [V].[Fe] PNXOJQQRXBVKEX-UHFFFAOYSA-N 0.000 description 10
- 230000003647 oxidation Effects 0.000 description 10
- 238000007254 oxidation reaction Methods 0.000 description 10
- 230000002572 peristaltic effect Effects 0.000 description 9
- 235000019422 polyvinyl alcohol Nutrition 0.000 description 9
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 8
- 229910052500 inorganic mineral Inorganic materials 0.000 description 8
- 235000010755 mineral Nutrition 0.000 description 8
- 239000011707 mineral Substances 0.000 description 8
- 238000004321 preservation Methods 0.000 description 8
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 7
- 238000004090 dissolution Methods 0.000 description 7
- 229910052751 metal Inorganic materials 0.000 description 7
- 239000002184 metal Substances 0.000 description 7
- 239000011734 sodium Substances 0.000 description 7
- 229910052708 sodium Inorganic materials 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 241000589925 Leptospirillum Species 0.000 description 5
- 239000010410 layer Substances 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 4
- 229910002651 NO3 Inorganic materials 0.000 description 4
- 241001134777 Sulfobacillus Species 0.000 description 4
- 241000605118 Thiobacillus Species 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 239000000654 additive Substances 0.000 description 4
- 230000000996 additive effect Effects 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 239000011805 ball Substances 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 4
- 239000004327 boric acid Substances 0.000 description 4
- 239000011575 calcium Substances 0.000 description 4
- 239000002131 composite material Substances 0.000 description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- 241000605272 Acidithiobacillus thiooxidans Species 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- 206010013786 Dry skin Diseases 0.000 description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 3
- 238000005273 aeration Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
- 238000013329 compounding Methods 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 229910052564 epsomite Inorganic materials 0.000 description 3
- 230000009123 feedback regulation Effects 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 3
- 239000006194 liquid suspension Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- -1 polyethylene Polymers 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- 230000001376 precipitating effect Effects 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 238000012549 training Methods 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 230000032258 transport Effects 0.000 description 3
- 239000002351 wastewater Substances 0.000 description 3
- 238000004065 wastewater treatment Methods 0.000 description 3
- 241000726121 Acidianus Species 0.000 description 2
- 241000521593 Acidimicrobium Species 0.000 description 2
- 241000266272 Acidithiobacillus Species 0.000 description 2
- 241001147780 Alicyclobacillus Species 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241001280345 Ferroplasma Species 0.000 description 2
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 description 2
- 241000255640 Loa loa Species 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- 241000202898 Ureaplasma Species 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000012050 conventional carrier Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 230000000855 fungicidal effect Effects 0.000 description 2
- 239000000417 fungicide Substances 0.000 description 2
- 229910052935 jarosite Inorganic materials 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000004114 suspension culture Methods 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 231100000033 toxigenic Toxicity 0.000 description 2
- 230000001551 toxigenic effect Effects 0.000 description 2
- 239000003981 vehicle Substances 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 241000605008 Spirillum Species 0.000 description 1
- DPSZMPUXFZOWAC-UHFFFAOYSA-N [S].S=O Chemical compound [S].S=O DPSZMPUXFZOWAC-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 150000001340 alkali metals Chemical class 0.000 description 1
- 229940037003 alum Drugs 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 231100000481 chemical toxicant Toxicity 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- TXKMVPPZCYKFAC-UHFFFAOYSA-N disulfur monoxide Inorganic materials O=S=S TXKMVPPZCYKFAC-UHFFFAOYSA-N 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000010406 interfacial reaction Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000005272 metallurgy Methods 0.000 description 1
- 239000011806 microball Substances 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000001473 noxious effect Effects 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- XTQHKBHJIVJGKJ-UHFFFAOYSA-N sulfur monoxide Chemical compound S=O XTQHKBHJIVJGKJ-UHFFFAOYSA-N 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
Abstract
The invention belongs to microorganisms to embed body field, specifically related to a kind of embedding body, preparation method and cultural method for loading leaching microbacteria, the embedding body for loading leaching microbacteria is the core-shell structure of central filler leaching microbacteria, nuclear structure is nutrient pill, nutrient pill is the particle formed after the embedding medium containing nutrient solution is crosslinked, and the nutrient solution is the culture solution without energy substance；Shell structure is that crosslinking agent is attached to the coated film formed after nutrient pill, and coated film is netted membrane structure；It is leaching microbacteria between the coated film and nutrient pill.The present invention solves leaching microbacteria, and the utilization of nutriment and energy substance consume unmatched problem during the cultivation process, overcome the problems such as difficult adaptation, easy poisoning, multithread of the leaching microbacteria in leaching environment are lost in bioleaching process, solve the problems such as being difficult to ensure hiding, difficult transport of leaching microbacteria, make the more convenient utilization of leaching microbacteria, leaching efficiency is improved.
Description
Technical field
The invention belongs to a kind of microorganisms to embed body field, be specifically related to a kind of embedding for loading leaching microbacteria
Body, preparation method and cultural method.
Background technique
Bioleaching refers to using leaching microbacteria from mineral etc. containing metal required for recycling in raw material metal, is one
Kind directly or indirectly acts on the method for dissolving out heavy metal from mineral or composite interstitial substance under acidic environment using microorganism,
Therefore also referred to as bioleaching.The microorganism for being generally used for bioleaching process belongs to bacterium domain or Archimycetes domain, and meets
Two primary conditions: they are acidophilic bacterias and are chemolithotrophies.Commonly used acidophilus leaching microbacteria has acidophilus
Thiobacillus (Acidithiobacillus spp.), acidocaldarius category (Alicyclobacillus spp.), sour germ category
(Acidimicrobium spp.), Sulfobacillus (Sulfobacillus spp.), sour Pseudomonas (Acidianus spp.),
Iron Ureaplasma (Ferroplasma spp.) and Leptospirillum (Leptospirillum spp.) etc..
Bioleaching is usually that mineral are accumulated or hoarded, and pouring sulfuric acid makes it keep being suitble to leaching microbacteria growth
Acid condition.Since leaching microbacteria is extreme acidophilic microorganism, suitable pH environment is 1.5~2.0, and many practical mines
Object Leaching Systems are increasingly complex, and not only pH value fluctuation range is big, and the toxic metal ions containing high concentration, and microorganism is such
It is difficult to grow in environment, or even can be dead；, and leaching microbacteria is to the mismatch of energy substance and nutrient utilization rate
Property, the cell viability of leaching microbacteria is not only influenced, it is unfavorable and dead because of environmental condition to will also result in many microorganisms.It is another
Aspect is usually that this quasi-microorganism is cultivated in open culturing system in biological metallurgy engineering, when microorganism grows into centainly
It often will appear the protozoan (such as eye worm) of a large amount of predation leaching microbacterias after concentration, in natural environment, once there is this
Class protozoan, cultivating system will be unbalance, this greatly reduces the survival volume and growth rate of leaching microbacteria, while cultivating
It will form adsorption and sedimentation and mineral surfaces characteristic changing that the associations product such as autunezite causes microorganism in the process, thus seriously
Affect the growth of microorganism and the leaching efficiency to mineral.
To solve the above-mentioned problems, microorganism is handled frequently with the mode of embedding, the currently used micro- life of embedding
The method of object is to mix microbial inoculum and occlusion vehicle, then the mixed liquor containing bacterium solution is added drop-wise in crosslinking agent, is handed over
Connection hardening forms the embedding structure containing microorganism.Patent if Chinese Patent Application No. is CN201510465105.9 discloses
A kind of complex micro organism fungicide and the preparation method and application thereof, includes the following steps, S1, bacillus is seeded in culture medium
Aerobic culture, obtains bacterium solution；S2, the bacterium solution is added in occlusion vehicle, obtains the mixed liquor containing bacterium solution, then contained by described in
The mixed liquor of bacterium solution is instilled into crosslinking agent dropwise, is carried out crosslinking hardening and is formed bacillus gel bacterium ball；S3, by the gemma
Bacillus gel bacterium ball carries out aerobic culture, obtains complex micro organism fungicide.Although such imbedded microbe can consolidate microorganism
It is scheduled in embedding body, but there are apparent differences for embedding surface layer and the supply of internal oxygen, embed internal portion and there is scarce/anaerobism
Environment will affect the growth of aerobic microbiological.And the embedding for iron-oxidizing bacteria, generally it will be considered that, if being embedded in inside,
Not only leaching microbacteria is difficult to obtain sufficient oxygen and carbon dioxide, and after ferrous ion enters embedding internal portion including
The ferrovanadium formed under portion's microbial action, be easy obstruction support voids, seriously affect nutriment release and microorganism after
Continuous regeneration can not carry out microorganism to continue culture.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of embedding body, preparation method and cultures for loading leaching microbacteria
Method overcomes the problems such as leaching microbacteria is to the difficult adaptation of leaching environment, easily poisoning, multithread mistake in bioleaching process,
The problems such as being difficult to ensure hiding, difficult transport of leaching microbacteria is solved, makes the more convenient utilization of leaching microbacteria, leaching efficiency is mentioned
It is high.
A kind of embedding body of load leaching microbacteria of the invention is the core-shell structure of central filler leaching microbacteria, core
Structure is nutrient pill, and nutrient pill is the particle formed after the embedding medium containing nutrient solution is crosslinked, and the nutrient solution is culture leaching mine
The culture solution without energy substance of microorganism；Shell structure is that crosslinking agent is attached to the coated film formed after nutrient pill, coated film
For netted membrane structure；It is leaching microbacteria between the coated film and nutrient pill.
Leaching microbacteria of the invention is preferably acidophilus leaching microbacteria, including thiobacillus ferrooxidans belong to
(Acidithiobacillus spp.), acidocaldarius category (Alicyclobacillus spp.), sour germ category
(Acidimicrobium spp.), Sulfobacillus (Sulfobacillus spp.), sour Pseudomonas (Acidianus spp.),
Iron Ureaplasma (Ferroplasma spp.) and Leptospirillum (Leptospirillum spp.) etc..
Leaching microbacteria of the invention is largely focused between coated film and nutrient pill, and a small amount of leaching microbacteria can seep
Thoroughly in coated film and nutrient pill, leaching microbacteria has relatively free space, will not block the hole of coated film and nutrient pill
Gap improves mass-transfer efficiency.
Nutriment is embedded in the internal layer of embedding body by the present invention, and outer layer then embeds leaching microbacteria, microorganism is embedded
In the spherical multilayer cross-linked structure (coated film) that toughness is high, toxicity is low, the gold of extraneous solution middle and high concentration can be not only reduced
Belong to the influence of ion or noxious material to leaching microbacteria, and embed nutrition composition slow release in body, after making embedding
Microorganism can be able to its bioactivity of surviving and keep for a long time, be convenient for its permanent preservation or long-distance transport, while reducing again
The additional loss of microorganism.The high activity of the high mass transfer ability and microorganism that embed body can enable it numerous in Primary culture
Grow speed faster, when practical application can reach required biomass within a short period of time.
Nutrient solution of the invention is the culture solution without containing energy substance, and energy substance is mainly ferrous ion, so battalion
Nutrient solution does not contain ferrous ion, it is preferred that the nutrient solution contains ammonium ion, potassium ion, magnesium ion, nitrate ion and calcium
Ion.Preferably, the nutrient solution includes 3~6g/L of ammonium sulfate, 0.5~2g/L of dipotassium hydrogen phosphate, 0.1~0.3g/ of potassium chloride
L, 0.5~2g/L of magnesium sulfate, 0.01~0.03g/L of calcium nitrate.The pH value of the nutrient solution is 1.5~2.0.Using above-mentioned nutrition
Liquid is conducive to the growth and breeding of leaching microbacteria.
The embedding medium is sodium alginate, preferably sodium alginate and polyvinyl alcohol mixture, more preferably glutaraldehyde, sea
Mosanom and polyvinyl alcohol mixture, wherein the volume fraction of glutaraldehyde is 1-1.5%.Preferably, sodium alginate and polyethylene
The mass ratio of alcohol is 1:10.Sodium alginate can make the immobilized spherule to be formed stronger after being sufficiently swollen, and polyvinyl alcohol is added
The balling-up of carrier can be improved, protects carrier, and polyvinyl alcohol is higher compared with sodium alginate intensity, it is ensured that immobilized spherule
Stability；Glutaraldehyde can improve embedding effect, extend the holding time.
Preferably, the crosslinking agent is saturation boric acid-calcium chloride solution added with magnesium sulfate, the quality volume of magnesium sulfate
Concentration is 0.3-0.5g/L, and magnesium sulfate can increase crosslinking agent toughness.Preferably, in 1L solution, typically contain 40g boric acid and
40g anhydrous calcium chloride.
The present invention also provides a kind of preparation methods of the embedding body of load leaching microbacteria, include the following steps, by nutrition
Ball is added to be collected by centrifugation after suspended with nutrient solution after leaching microbacteria in, after filtering, by above-mentioned attachment leaching microbacteria
Nutrition pellet is added in crosslinking agent, obtains the embedding body of the load leaching microbacteria；The preparation method of the nutrient pill is to incite somebody to action
Embedding medium containing nutrient solution is added drop-wise in crosslinking agent, and nutrient pill is formed after crosslinking.Leaching microbacteria bacterium after nutrient solution suspension
The bacterium of liquid dense preferably 3~5 × 109cells/mL。
After high concentration leaching microbacteria solution made of nutrient pill addition is suspended with nutrient solution, the automatic suction of leaching microbacteria
Attached ability is stronger, can automatically be adsorbed on the surface of nutrient pill, immerses in crosslinking agent after filtering above-mentioned substance, crosslinking agent exists
Its surface forms coated film, and leaching microbacteria is predominantly located between nutrient pill and coated film, and the leaching in nutrient pill and coated film
Mine microorganism is considerably less, can ignore.
The preparation method of embedding medium containing nutrient solution is that sodium alginate and polyvinyl alcohol are dissolved in nutrient solution respectively
In, then above two nutrient solution equal proportion is mixed, glutaraldehyde is added, obtains the embedding medium containing nutrient solution.
Sodium alginate and polyvinyl alcohol are separately dissolved in nutrient solution by the present invention, are then mixed, such method alginic acid
Sodium can sufficiently dissolve, and dissolution is more uniform.
The rate of addition that embedding medium containing nutrient solution is added drop-wise to crosslinking agent is 2-3 drops/sec.
The present invention also provides a kind of cultural method of embedding body culture leaching microbacteria using load leaching microbacteria, steps
It suddenly include that the embedding body for loading leaching microbacteria is added in the aqueous solution containing ferrous ion, regulates pH value (pH value 1.5-
2.0), bored exposure culture.Aqueous solution containing ferrous ion is preferably ferrous sulfate aqueous solution.
The invention has the advantages that
1, the present invention provides a kind of method of fixed extreme acidophilus leaching microbacteria of embedding, is embedded and is soaked using immobilization technology
The technology of mine microorganism can overcome leaching microbacteria difficult culture under conventional conditions, easily death, high investment, low leach it is scarce
Point.Under embedding conditions of the invention, nutriment slow release, leaching microbacteria can realize long period culture；And microballoon
With certain moisture, convenient for the permanent preservation and transport of thallus；Microorganism is fixed on carrier surface or inside, to enhance embedding
The activity and mass transfer ability of fixed microorganism carrier, convenient for the growth and release of microorganism.This kind of embedded immobilization technology energy
Effective flora is fixed, can quickly improve the biomass of leaching microbacteria, and keep the activity of efficient bacterium, it can also
Different leaching microbacterias and nutriment are mixed to form a complex, the effective composite flora embedding of leaching microbacteria is formed
Body greatly improves treatment effeciency to play more preferably bioleaching effect, reduces processing cost.Such embedding method to wrap
It buries microbial quality to stablize, diffusion is excellent, can industrialization promotion use conducive to the commercialization of thallus.
2, embedding body produced by the present invention, inside is mineral nutrition needed for leaching microbacteria, but microorganism growth needs
It wants extraneous to provide again energy substance (ferrous ion), embedding body high nutrition, the few energy and internal hyperosmosis are conducive to the leaching micro- life of mine
The permanent preservation of object.Aqueous solution pH need to be only adjusted to 1.5-2.0 in culture, the energy containing ferrous ion is added in the solution
Substance, such as ferrous sulfate or containing ferrous treating material or containing the circulation fluid of ferrous ion.Directly add after preparing solution
Add this embedding body can Primary culture, not only enormously simplify culture process, also significantly increase microorganism growth and breeding speed
Degree, hence it is evident that shorten the cultivation cycle for reaching required biomass.Incubation substantially increases extreme acidophilus leaching microbacteria
Cultivate rate and survival volume.Embedding structure of the invention consumes microbial nutrition substance and energy substance can be effective
Match, the two depletion rate is adapted, nutriment slow release, meets microorganism demand, generation ammonia nitrogen waste water is less, while energy
Source substance sufficiently can be changed into a large amount of ferric ion by ferrous ion, improve the oxidation-reduction potential in cultivating system
Mine effect is more preferably soaked to play, nutriment consumption is slower, can effectively meet the growth needs of microorganism long-term cultivation.
3, due to the ammonium ion (NH in nutrient solution4 +) and potassium ion (K+) easily with ferric ion (Fe3+) combine shape
At yellow ammonium ferrovanadium [NH4Fe3(SO4)2(OH)6] and autunezite [KFe3(SO4)2(OH)6].Especially in microbial leaching system,
Fe under the action of microorganism in solution2+It is oxidized to Fe3+Afterwards, certain temperature, acidity and have ammonium ion or alkali metal from
Under the conditions of existing for sub, jarosite can be formed, and microorganism accelerates the reaction.And the invention enables ammonium ions and potassium
Ion is embedded in inside, does not contact largely with extraneous iron ion, reduces a large amount of generations of ferrovanadium, decreases raw to microactuator suspension
The adsorption and sedimentation of object improves the microorganism yield of unit dose, more conducively the microorganism total amount in raising system and output effect
Rate.
4, due to the iron oxidation efficiency about 0.2-2g/Lh in leaching microbacteria incubation, ferrous ion is to microorganism
For be energy substance, and it is usually in 48-168h that the growth of every batch of microorganism, which reaches high-biomass, therefore Microbial Iron aoxidizes
Rate and high-biomass seriously mismatch, and the nutrition pellet in the present invention can make nutriment slow release, can not only prolong
Long cultivation cycle, but also iron oxidation rate and biology are flux matched, on the one hand ferrous ion is used as the energy of microorganism to support substance, separately
On the one hand the trivalent ferrous solution after aoxidizing can be used as leaching agent use, can reduce the input cost of nutriment.
5, embedding method of the invention overcomes is also easy to produce a large amount of autunezite in traditional microbial cultivation process,
The problem for causing the iron concentration of bioavailable low；The ferrovanadium passivating film for avoiding generation seriously affects mineral or extract
The precipitating of the leaching and generation of metal ion takes away a part of metal or causes the migration of metal and convert and hinder metal in material
Extraction the problems such as generation；Simultaneously solve by bioavailable iron concentration reduce caused by interfacial reaction it is slack-off,
The problems such as Deal with Time extends, disposition is not thorough.
6, cultural method of the invention is not only simple and easy, save the cost, it is thus also avoided that nutriment is to Leaching Systems
Pollution conducive to the metal separation in solution after leaching, and alleviates the discharge burden of nitrogen in waste water, phosphorus, potassium；And reduce iron
The generation of alum effectively prevents siderotil absorption suspension thalline, alleviates subsequent wastewater treatment burden, improves the biology training of system
Support efficiency and yield.
7, the embedding method of the invention to extreme acidophilus leaching microbacteria can not only be directly exposed to avoid microorganism
Change cell viability in extreme acidic's medium (pH value is lower than 1.0), toxic mineral environment can also be directly contacted to avoid it
In (such as containing the raffinate of toxic chemical substance), being poisoned to death for microorganism is avoided, while in turn avoiding leaching microbacteria by water
The phagocytosis of the protozoans such as the eye worm in solution solves the problems, such as that ore-leaching bacteria is easily dead in the solution, is difficult to ensure and deposits.
8, imbedded microbe ball produced by the present invention can be sprinkled upon in right amount on material to be leached or be mixed with material to be leached
It places, microorganism can constantly be grown using the energy substance of nutriment and the external world in embedding body, to realize in material
The leaching of metal.It can prevent from largely pouring environmental pollution brought by biochemical lixivium and wastage of material.
9, embedding method provided by the invention can embed the compound bacteria of different specific leaching microbacterias, can be according to expectation
Contain any microorganism or microbial population encapsulated with arbitrary ratio.The Bioleaching effect of these compound bacterias compares single leaching
Mine microorganism can play more preferably bioleaching effect.
10, multilayer of the invention embeds structure, and internal there is certain density nutriment can form certain buffering
System, avoids the fluctuation of pH value, and the reticular structure of embedding can effectively obstruct protozoan to acidophilus leaching microbacteria
Predation, provides a good growing environment for leaching microbacteria, improves the culture efficiency of microorganism.
Detailed description of the invention
Fig. 1 is the preparation flow schematic diagram of nutrient pill of the invention.
Fig. 2 is structural schematic diagram of the invention.
Fig. 3 is the shape graph of the embedding body of load leaching microbacteria of the invention.
Fig. 4 is that the embedding body of load leaching microbacteria of the invention expands the aspect graph after culture.
Fig. 5 is that the expansion culture effect of the present invention and comparative example 2 compare figure.Left side is the expansion after present invention culture 5 days
Cultured products, right side are the expansion cultured products after comparative example 2 is cultivated 5 days.
Fig. 6 is that comparative example 2 of the present invention expands the effect picture after culture 20 days.
Fig. 7 is two batches of culture effect figures of the method for the present invention, is followed successively by two batches of culture starting points, two batches of cultures two from left to right
It, two batches of cultures, five days effect pictures.
1 embedding medium, 2 peristaltic pumps, 3 suction tubes, 4 crosslinking agents, 5 nutrient pills, 6 leaching microbacterias, 7 coated films in the figure.
Specific embodiment
Embodiment 1
A kind of embedding method of extreme acidophilus leaching microbacteria.Specific step is as follows:
(1) configuration of nutrient solution
Ammonium sulfate ((NH4)2SO4) 4g/L, dipotassium hydrogen phosphate (K2HPO4) 1g/L, potassium chloride (KCl) 0.2g/L, magnesium sulfate
(MgSO4·7H2O) 1g/L and calcium nitrate (Ca (NO3)2) 0.02g/L, the distilled water of corresponding amount is added, with 98% concentrated sulfuric acid
(H2SO4) adjust pH to 2.0 or so.
(2) prepared by crosslinking agent
Calcium chloride-boric acid (CaCl of 4% (w/v) is configured with distilled water2-H3BO3) solution, i.e., with distilled water by 40g boric acid
40g anhydrous calcium chloride is added after dissolution completely, is settled to 1L with distilled water after dissolution, crosslinking agent is made.In every liter of crosslinking agent
Magnesium sulfate (MgSO is added4·7H2O) 0.3g, to increase crosslinking agent toughness.
(3) prepared by embedding medium
Embedding medium is the mixed liquor of the sodium alginate-polyvinyl alcohol (SA-PVA) of compounding.
It weighs appropriate sodium alginate and is swollen 7h in nutrient solution, dissolved in 85 DEG C of hot water completely, it is 2% that content, which is made,
Sodium alginate soln.The poly-vinyl alcohol solution for being 20% with nutrient solution content.Two kinds of solution are mixed in equal volume, must be contained
Amount is respectively 1% and 10% sodium alginate-polyvinyl alcohol (SA-PVA) mixed liquor, and glutaraldehyde (GA) is added in mixed liquor to be made
Its volume fraction reaches 1.5%, and embedding medium is made.
(4) prepared by nutrition pellet
Legal system nutrition pellet is oozed using peristaltic pump.It is above-mentioned with rubber tube (internal diameter Φ=2mm) suction of peristaltic pump one end
Nutrient solution mixed liquor, the other end instill in above-mentioned crosslinking agent, coutroi velocity, so that it is 2~3/second that drop, which oozes speed, stand
1h forms nutrition pellet particle, then by the 30 DEG C of dryings of nutrition pellet.Such production method makes nutrition pellet dimensionally stable
Property it is high, it is good to form effect.
(5) leaching microbacteria embeds
Thiobacillus ferrooxidans (Acidithiobacillus ferrooxidans, A.f) culture is extremely grown into logarithmic phase,
Thalline were collected by centrifugation, with high concentration nutritional liquid suspension thalline, makes its bacterial concentration up to 3~5 × 109Seed is made in cells/mL
Liquid.
Nutrition pellet is uniformly poured slowly into above-mentioned seed liquor, so that microorganism adsorption is wrapped in nutrition pellet outer layer,
The nutrition pellet for being covered with microorganism is formed, is retransferred after being filtered out liquid into 10min stable in crosslinking agent, formation is covered with coating
The biological microsphere of film completes microorganism embedding, obtains the embedding body of load leaching microbacteria.It is specifically shown in Fig. 3.
4 DEG C of preservations after embedding body surface solution are drained, are directly added directly it according to 5% ratio when needing to cultivate
Aeration culture can be started into the solution of pH=1.5~2.0 of the ferrous ion containing 6g/L.Aspect graph is as shown in Figure 4 after culture.
(1) continuous overflow training method is used
Specific cultural method are as follows: swept volume be 10L continuous overflow cultivation reactor in, by A.f bacterium carrier with
5% additive amount is added in the ferrous sulfate solution of 9.5L, and bored to expose 7 days, solution thoroughly reddens, and current potential reaches 600mV or more,
Check the concentration of the microorganism in solution, the microorganism concn in solution reaches 5 × 107When a/mL, start continuous culture
Mode.The feed liquor continuously cultivated is the acid ferrous sulfate solution (pH=2.0) of 6g/L, and feed liquor flow velocity is by liquid outlet current potential
It is no to come feedback regulation (will increase more than 600mV flow velocity, on the contrary then slow down) more than 600mV.
Culture effect: the continuous culture of starting obtains 600mV or more ferrous sulfate microbial solution after 60 days is total about
5m3, be averaged ferrous oxidation rate 2.28g/ (Lh), the ammonia nitrogen 20mg/L in solution, total phosphorus 0.08mg/L, and ferric ion is dense
5.82g/L is spent, ammonia nitrogen, total phosphorus, which contain, in liquid can reach direct emission standard.
(2) batch culture mode is used
After culture solution reaches required biomass, take out cultivating system 90% microbial solution, retain embedding body and
10% bacterium solution continues to add ferrous sulfate aqueous solution containing 6g/L (pH=1.8), is free of nutritional ingredient, bored to expose 7 days.
Culture effect: microorganism starts raised growth after bored exposure 5 days, and ferrous ion starts quickly to aoxidize, and cultivates body after 7 days
System reaches required biomass, average iron oxidation rate 1.82g/ (L/8), the ammonia nitrogen 16mg/L in solution, total phosphorus 0.05mg/L, and three
Valence iron concentration 5.32g/L, ammonia nitrogen, total phosphorus can reach direct emission standard in liquid.It is specifically shown in Fig. 7, it can be seen that even if
It cultivates in batches, microorganism remains to rapid, high volume growth, illustrates that embedding body still has sufficient nutrient to support microorganism, it was demonstrated that embedding
Nutriment can actually realize slow release in body, and microorganism is made to can make full use of these nutriments, to reduce original
Material investment.
Embodiment 2
A kind of embedding method of extreme acidophilus leaching microbacteria.Specific step is as follows:
(1) configuration of nutrient solution
Ammonium sulfate ((NH4)2SO4) 5g/L, dipotassium hydrogen phosphate (K2HPO4) 1g/L, potassium chloride (KCl) 0.3g/L, magnesium sulfate
(MgSO4·7H2O) 2g/L and calcium nitrate (Ca (NO3)2) 0.03g/L, the distilled water of corresponding amount is added, with 98% concentrated sulfuric acid
(H2SO4) adjust pH to 1.5~2.0.
(2) prepared by crosslinking agent
Calcium chloride-boric acid (CaCl of 4% (w/v) is configured with distilled water2-H3BO3) solution, i.e., with distilled water by 40g boric acid
40g anhydrous calcium chloride is added after dissolution completely, is settled to 1L with distilled water after dissolution, crosslinking agent is made.In every liter of crosslinking agent
Magnesium sulfate (MgSO is added4·7H2O) 0.3g, to increase crosslinking agent toughness.
(3) prepared by embedding medium
Embedding medium is the mixed liquor with the sodium alginate-polyvinyl alcohol (SA-PVA) of compounding.
It weighs appropriate sodium alginate and is swollen 8h in nutrient solution, dissolved in 80 DEG C of hot water completely, it is 2% that content, which is made,
Sodium alginate soln.The poly-vinyl alcohol solution for being 20% with nutrient solution content.Two kinds of solution are mixed in equal volume, must be contained
Amount is respectively 1% and 10% sodium alginate-polyvinyl alcohol (SA-PVA) mixed liquor, and embedding medium is made.
(4) prepared by nutrition pellet
Legal system nutrition pellet is oozed using peristaltic pump.It is above-mentioned with rubber tube (internal diameter Φ=2mm) suction of peristaltic pump one end
Nutrient solution mixed liquor, the other end instill in above-mentioned crosslinking agent, coutroi velocity, so that it is 2~3/second that drop, which oozes speed, stand
1h forms nutrition pellet particle, by the 30 DEG C of dryings of nutrition pellet.Such production method makes nutrition pellet dimensional stability
It is good to form effect for height.
(5) leaching microbacteria embeds
By Thiobacillus ferrooxidans (Acidithiobacillus ferrooxidans) and Thiobacillus thioxidans
To growth logarithmic phase, thalline were collected by centrifugation, outstanding with high concentration nutritional liquid for (Acidithiobacillus thiooxidans) culture
Floating thallus, makes its bacterial concentration up to 3~5 × 109Seed liquor is made in cells/mL.
Nutrition pellet is uniformly poured slowly into upper seed liquor, so that microorganism is wrapped in nutrition pellet outer layer, formation is covered
There is the nutrition pellet of microorganism, retransfers after being filtered out liquid into 10min stable in crosslinking agent, form the life for being covered with coated film
Object microballoon completes microorganism embedding, obtains the embedding body of load leaching microbacteria.
4 DEG C of preservations after embedding body surface solution are drained, directly directly throw it according to 10% ratio when needing to cultivate
Aeration culture can be started by being added in the solution of pH=1.5~2.0 of the ferrous ion containing 6g/L.
Specific cultural method are as follows: in the continuous overflow cultivation reactor that swept volume is 50L, by embedding body with 10%
Additive amount be added in the ferrous sulfate solution of 45L, it is bored expose 8 days, solution thoroughly reddens, and current potential reaches 600mV or more, check
The concentration of microorganism in solution, the microorganism concn in solution reach 5 × 107When a/mL, start continuous overflow mode
Culture.The feed liquor continuously cultivated is the acid ferrous sulfate solution (pH1.8) of 8g/L, and whether feed liquor flow velocity passes through liquid outlet current potential
Come feedback regulation (will increase more than 600mV flow velocity, on the contrary then can slow down) more than 600mV.
Culture effect: the continuous culture of starting obtains 600mV or more ferrous sulfate microbial solution after 60 days is total about
35m3, average ferrous oxidation rate 2.31g/ (Lh), the ammonia nitrogen 17.7mg/L in solution, total phosphorus 0.09mg/L, ferric iron from
Sub- concentration 5.93g/L, liquid is up to direct emission standard.
Embodiment 3
A kind of embedding method of extreme acidophilus leaching microbacteria.Specific step is as follows:
(1) configuration of nutrient solution
Ammonium sulfate ((NH4)2SO4) 0.5~5g/L, dipotassium hydrogen phosphate (K2HPO4) 0.2~2g/L, potassium chloride (KCl) 0.1~
0.2g/L and calcium nitrate (Ca (NO3)2) 0.03g/L, the distilled water of corresponding amount is added, with sulfuric acid (H2SO4) adjust pH to 3.0.
(2) prepared by crosslinking agent
Calcium chloride-boric acid (CaCl of 4% (w/v) is configured with distilled water2-H3BO3) solution, i.e., with distilled water by 40g boric acid
40g anhydrous calcium chloride is added after dissolution completely, is settled to 1L with distilled water after dissolution, crosslinking agent is made.In every liter of crosslinking agent
Magnesium sulfate (MgSO is added4·7H2O) 0.3~0.5g, to increase crosslinking agent toughness.
(3) prepared by embedding medium
Embedding medium is the mixed liquor with the sodium alginate-polyvinyl alcohol (SA-PVA) of compounding.
It weighs appropriate sodium alginate and is swollen 6~8h in nutrient solution, dissolved in 80~90 DEG C of hot water completely, be made and contain
The sodium alginate soln that amount is 2%.The poly-vinyl alcohol solution for being 20% with nutrient solution content.Two kinds of solution are mixed in equal volume
It closes, obtains sodium alginate-polyvinyl alcohol (SA-PVA) mixed liquor that content is respectively 1% and 10%.
(4) prepared by nutrition pellet
Legal system nutrition pellet is oozed using peristaltic pump.It is above-mentioned with rubber tube (internal diameter Φ=2mm) suction of peristaltic pump one end
Nutrient solution mixed liquor, the other end instill in above-mentioned crosslinking agent, coutroi velocity, so that it is 2~3/second that drop, which oozes speed, stand
1h forms nutrition pellet particle, by the 30 DEG C of dryings of nutrition pellet.Such production method makes nutrition pellet dimensional stability
It is good to form effect for height.
(5) leaching microbacteria embeds
By leaching microbacteria, such as Thiobacillus ferrooxidans (Acidithiobacillus ferrooxidans), sulfur oxide
Thiobacillus (Acidithiobacillus thiooxidans), ferrous oxide hook end spirillum (Leptospirillum
) etc. ferriphilum to logarithmic growth phase, thalline were collected by centrifugation, with high concentration nutritional liquid suspension thalline and mixes preparation for culture
At leaching microbacteria composite bacteria liquid, make its bacterial concentration up to 3~5 × 109Seed liquor is made in cells/mL or so.
Nutrition pellet is uniformly poured slowly into upper seed liquor, so that microorganism is wrapped in nutrition pellet outer layer, formation is covered
There is the nutrition pellet of microorganism, retransfers after being filtered out liquid into 5min stable in crosslinking agent, form the life for being covered with coated film
Object microballoon completes microorganism embedding, obtains the embedding body of load leaching microbacteria.
4 DEG C of preservations after embedding body surface solution are drained, directly directly throw it according to 15% ratio when needing to cultivate
Aeration culture can be started by being added in the solution of pH=1.5~2.0 of the ferrous ion containing 10g/L.
Specific cultural method are as follows: swept volume be 1m3Continuous overflow cultivation reactor in, will embedding body with 15%
Additive amount be added in the ferrous sulfate solution of 850L, it is bored expose 7 days, solution thoroughly reddens, and current potential reaches 600mV or more, inspection
The concentration of the microorganism in solution is looked into, the microorganism concn in solution reaches 5 × 107When a/mL, start continuous overflow mould
Formula culture.The feed liquor continuously cultivated is the acid ferrous sulfate solution (pH2.0) of 8g/L, and feed liquor flow velocity is by liquid outlet current potential
It is no to come feedback regulation (will increase more than 600mV flow velocity, on the contrary then slow down) more than 600mV.
Culture effect: the continuous culture of starting obtains 600mV or more ferrous sulfate microbial solution after 60 days is total about
10m3, average ferrous oxidation rate 2.30g/ (Lh), the ammonia nitrogen 15.87mg/L in solution, total phosphorus 0.08mg/L, ferric iron from
Sub- concentration 6.97g/L.
It under the same terms, is continuously cultivated using suspension training method, obtains solution about 2.5m after culture in 60 days3, sub-
Iron oxidation rate about 0.33g/L, ammonia nitrogen 635.6mg/L, total phosphorus 88.29mg/L, ferric ion 1.76g/L in solution, greatly
It measures ferrovanadium to generate, ammonia nitrogen, total phosphorus contain considerably beyond discharge standard.
It under the same terms, is continuously cultivated using 10% filler, obtains within 60 days solution about 2m3, average ferrous oxidation rate
About 0.842g/ (Lh), ammonia nitrogen 620mg/L, total phosphorus 85mg/L in ferric ion 1.96g/L solution, ammonia nitrogen, total phosphorus content
Considerably beyond discharge standard.
1, conventional suspension culture is compared with the present invention
The traditional cultural method of leaching microbacteria is to need first to configure mass propgation based sols according to culture medium requirement, then connect
Enter leaching microbacteria seed liquor, controls temperature, pH Primary culture under suitable conditions, cultivation cycle was at one week or more.And
Added a large amount of ferrous ion obtains energy substance as microorganism in culture medium, and the rate being oxidized is exceedingly fast, and microorganism
It is slower to the utilization rate of inorganic salts, therefore the inorganic salts being directly exposed in culture medium and ferric ion is caused to be formed largely
Autunezite and yellow ammonium ferrovanadium, not only cause a large amount of wastes of culture medium raw material, increase toxigenic capacity, and microorganism
It can largely adhere to or be embedded in precipitating, be substantially reduced biomass, it is difficult to by microculture to requirement.Containing height
The microorganism of concentration inorganic salts culture solution investment leaching mine using when necessarily to will cause serious nitrogen, phosphorus, potassium content exceeded, cause
Serious contaminated wastewater, therefore the cultural method of traditional leaching microbacteria can not only make cultivation cycle extend, toxigenic capacity
Increase, leaching effect decline, also can bring very heavy burden to the wastewater treatment in later period, or even will increase the risk of potential pollution.
And cultural method of the invention is used, due to being different from batch experiments, nutriment and cell after microculture
It is taken away in batches with material, compared to traditional cultural method, the nutriments such as inorganic salts in the nutrition pellet in the present invention
Slow release can extend the cycles consumed of nutriment, significantly improve the utilization efficiency of nutriment.It obtains same amount of micro-
Biology, the consumption of nutriment reduce 3~5 times than conventional method, not only avoid ammonium sulfate and potassium chloride of high concentration etc. with
Ferric ion forms a large amount of yellow ammonium ferrovanadiums and autunezite, and reduces raw material investment, reduces production cost.Moreover,
It since nutriment is included in nutrition pellet, will not directly largely be exposed in solution, therefore can be significantly reduced even not
Water body nitrogen, phosphorus, potassium pollution are generated, can effectively mitigate later period sewage treatment burden, mitigate environmental protection pressure.
2, conventional carriers culture is compared with the present invention
Traditional microbe carrier mainly utilizes microorganism in carrier surface or reticular structure attachment to form biomembrane,
This kind of carrier adds support material after needing to cultivate a large amount of high density microorganism seed liquid during the preparation process in advance, for a long time
Biomembrane could be formed in carrier surface after mixed culture, then carefully take out overlay film carrier, need first to match in Primary culture
Culture medium has been set, then has added carrier, has controlled suitable condition with Primary culture.It is easy in this kind of carrier preparation process because of support material
Difference cause the growth of microorganism to be suppressed, also having when adding carrier Primary culture can due to the sudden change of external environment
Just start to grow after the lag phase for needing the long period.Microorganism also can be because of water shortage and battalion in the preservation and transportational process of carrier
It supports substance and mortality or falls off, therefore, it is difficult to cultivate required biomass within a short period of time.
Microorganism embedding method of the invention overcomes the deficiency of traditional biological carrier, not only the letter of microorganism cover process
It is single, and ferrous or iron-bearing material, the automatic long period of embedding body added need to only be added in aqueous solution in Primary culture and mention
For nutriment needed for microorganism growth, and there is no inhibition for the embedding body of the cover microorganism added and aqueous solution
Phenomenon, microorganism can instant growth and breeding, therefore biggish biomass can be reached in a short time.In addition to this nutrition is micro-
Ball provides moisture and nutriment to the microorganism of cover, it is ensured that and microorganism can survive for a long time and maintain greater activity,
The long-time preservation of strain may be implemented.
Through the present invention in comparative example compared with conventional suspension culture and conventional carriers culture, using load of the invention
The biomass that body not only obtains in the same time is more, but also consumption nutrient raw material is few, contained ammonia nitrogen, total phosphorus in biological solution
Seldom, the pressure of subsequent wastewater treatment is alleviated, it is a kind of saving time that these advantages, which demonstrate the present invention, reduces cost and ring
The microorganism embedding method of guarantor.
The present invention is compared with conventional method, other than having the above advantage, also have retention cycle is long, Primary culture is fast,
The advantages that leaching effect is good.
Comparative example 1
This comparative example 1 is compared with embodiment 2, and step (1)-(3) are identical, and subsequent step is step (4), i.e. leaching microbacteria
Embedding.
Step (4): by Thiobacillus ferrooxidans (Acidithiobacillus ferrooxidans) and sulfur oxide sulphur bar
Bacterium (Acidithiobacillus thiooxidans) culture is to growth logarithmic phase, and thalline were collected by centrifugation, with high concentration nutritional liquid
Suspension thalline makes its bacterial concentration up to 3~5 × 109Seed liquor is made in cells/mL.
Seed liquor and embedding medium equivalent (seed liquor: embedding medium (v/v)=1:1) are stood into 30min after mixing, are made
Embed mixed liquor.
Legal system embedding body is oozed using peristaltic pump.Above-mentioned packet is aspirated with the rubber tube (internal diameter Φ=2mm) of peristaltic pump one end
Mixed liquor is buried, the other end instills in the mixed liquor of above-mentioned calcium chloride-boric acid, coutroi velocity, so that it is 2~3 that drop, which oozes speed,
A/second, embedding body stand 1h after being formed.The composite flora that Thiobacillus ferrooxidans and Thiobacillus thioxidans is made embeds body.
For cultural method with embodiment 2, difference is that the additive amount for embedding body is 15%.(embodiment 2 is 10%)
Culture effect: it is such microorganism be embedded in internal method to reach the required biomass time greatly prolong.
The continuous culture of starting obtains 600mV or more ferrous sulfate microbial solution after 60 days amounts to about 2m3, average ferrous oxidation rate
0.89g/ (Lh), ammonia nitrogen 12mg/L, total phosphorus 0.07mg/L, ferric ion concentration 5.32g/L in solution.
Comparative example 2
Comparative example 2 is with comparative example 1, and difference is cultural method difference, and the cultural method of comparative example 2 is to need to cultivate
When directly it is added directly in the culture solution of the ferrous ion containing 6g/L (pH=1.5~2.0) according to 15% ratio, bored exposure
Culture.Contain in culture solution: ammonium sulfate ((NH4)2SO4) 4g/L, dipotassium hydrogen phosphate (K2HPO4) 1g/L, potassium chloride (KCl) 0.2g/
L, magnesium sulfate (MgSO4·7H2O) 1g/L and calcium nitrate (Ca (NO3)2)0.02g/L。
Culture effect: the continuous culture of starting obtains 600mV or more ferrous sulfate microbial solution after 60 days is total about
35m3, average ferrous oxidation rate 0.89g/ (Lh), the ammonia nitrogen 565.2mg/L in solution, total phosphorus 78.3mg/L, ferric iron from
Sub- concentration 0.58g/L.
The comparison for expanding cultured products is as shown in Figure 5, it can be seen that connects under embedding method of the invention only 5%
(5 days) solution can be made to redden and (show that microorganism raised growth, ferric ion largely generate) in a short time when kind amount, reached
To larger biomass, and bacterium solution is limpid, and without the generation of the precipitatings such as ferrovanadium, and existing mixing embedding method is almost without life
Object amount generates, and liquid is muddy, illustrates to form more ferrovanadium on embedding body.This illustrates that mixing embedding seriously affects microorganism
Growth and release, and ferrovanadium generation cannot be prevented.Therefore has the not applicable embedding with leaching microbacteria of mixing embedding method.
As shown in fig. 6, effect picture of the comparative example 2 after continuous culture 20 days.Compare Fig. 5 and Fig. 6 it is found that mixing embedded
Although mode can star culture, but the incubation time for reaching required biomass is that 3-5 times of the time of the invention could be real
Biomass needed for existing present invention culture, cultivation cycle greatly prolong.
The color of effect picture after culture of the invention is deeper, and ferric ion is more in expression system, and biomass is more,
Ferrovanadium is fewer.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, the variation or replacement that can be readily occurred in,
It should be covered by the protection scope of the present invention.Therefore, protection scope of the present invention should be with the range of claims
It is quasi-.
Claims (10)
1. a kind of embedding body for loading leaching microbacteria, characterized in that be the core-shell structure of central filler leaching microbacteria, core knot
Structure is nutrient pill, and nutrient pill is the particle formed after the embedding medium containing nutrient solution is crosslinked, and the nutrient solution is that culture leaching mine is micro-
The culture solution without energy substance of biology；Shell structure is that crosslinking agent is attached to the coated film formed after nutrient pill, and coated film is
Netted membrane structure；It is leaching microbacteria between the coated film and nutrient pill.
2. as described in claim 1 load leaching microbacteria embedding body, characterized in that the nutrient solution contain ammonium root from
Son, potassium ion, magnesium ion, nitrate ion and calcium ion.
3. the embedding body of load leaching microbacteria as claimed in claim 2, characterized in that the nutrient solution includes ammonium sulfate 3
~6g/L, 0.5~2g/L of dipotassium hydrogen phosphate, 0.1~0.3g/L of potassium chloride, 0.5~2g/L of magnesium sulfate, calcium nitrate 0.01~
0.03g/L。
4. the embedding body of load leaching microbacteria as claimed in claim 2 or claim 3, characterized in that the pH value of the nutrient solution is
1.5~2.0.
5. the embedding body of load leaching microbacteria as described in claim 1, characterized in that the embedding medium is glutaraldehyde, sea
Mosanom and polyvinyl alcohol mixture, wherein the volume fraction of glutaraldehyde is 1-1.5%.
6. the embedding body of load leaching microbacteria as claimed in claim 5, characterized in that in the embedding medium, sodium alginate
Mass ratio with polyvinyl alcohol is 1:10.
7. the embedding body of load leaching microbacteria as described in claim 1, characterized in that the crosslinking agent is added with sulfuric acid
Saturation boric acid-calcium chloride solution of magnesium, the mass-volume concentration of magnesium sulfate are 0.3-0.5g/L.
8. a kind of preparation method for the embedding body for loading leaching microbacteria as described in claim any one of 1-7, characterized in that packet
Following steps are included, nutrient pill are added in the leaching microbacteria after being suspended after being collected by centrifugation with nutrient solution, it, will be above-mentioned after filtering
The nutrient pill of attachment leaching microbacteria is added in crosslinking agent, obtains the embedding body of load leaching microbacteria；The nutrient pill
Preparation method is that the embedding medium containing nutrient solution is added drop-wise in crosslinking agent, forms nutrient pill after crosslinking.
9. preparation method as claimed in claim 8, characterized in that the preparation method of the embedding medium containing nutrient solution is, will be extra large
Mosanom and polyvinyl alcohol are dissolved in nutrient solution respectively, then mix above two nutrient solution, and glutaraldehyde is added, is contained
The embedding medium of nutritious liquid.
10. a kind of using the embedding body culture leaching microbacteria of load leaching microbacteria as described in claim any one of 1-7
Cultural method, characterized in that the embedding body for loading leaching microbacteria is added in the aqueous solution containing ferrous ion, regulates pH
Value, bored exposure culture, culture to suspension cell concentration reach 1 × 108Start continuous culture when cell/mL or more.
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