CN115369053B - Azo dye degrading bacterium and application thereof - Google Patents
Azo dye degrading bacterium and application thereof Download PDFInfo
- Publication number
- CN115369053B CN115369053B CN202210654954.9A CN202210654954A CN115369053B CN 115369053 B CN115369053 B CN 115369053B CN 202210654954 A CN202210654954 A CN 202210654954A CN 115369053 B CN115369053 B CN 115369053B
- Authority
- CN
- China
- Prior art keywords
- enterococcus faecalis
- azo dye
- azo
- dye
- azo dyes
- 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.)
- Active
Links
- 239000000987 azo dye Substances 0.000 title claims abstract description 155
- 241000894006 Bacteria Species 0.000 title claims abstract description 27
- 230000000593 degrading effect Effects 0.000 title claims abstract description 22
- 241000194032 Enterococcus faecalis Species 0.000 claims abstract description 122
- 229940032049 enterococcus faecalis Drugs 0.000 claims abstract description 114
- 239000002351 wastewater Substances 0.000 claims abstract description 19
- 238000012258 culturing Methods 0.000 claims description 12
- 238000011081 inoculation Methods 0.000 claims description 6
- 238000004458 analytical method Methods 0.000 abstract description 19
- 230000000694 effects Effects 0.000 abstract description 19
- 231100000419 toxicity Toxicity 0.000 abstract description 11
- 230000001988 toxicity Effects 0.000 abstract description 11
- 238000006065 biodegradation reaction Methods 0.000 abstract description 9
- 229910001385 heavy metal Inorganic materials 0.000 abstract description 7
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 abstract description 6
- 150000001875 compounds Chemical class 0.000 abstract description 5
- 238000001784 detoxification Methods 0.000 abstract description 4
- 231100000053 low toxicity Toxicity 0.000 abstract description 4
- 150000003839 salts Chemical class 0.000 abstract description 4
- 239000003513 alkali Substances 0.000 abstract description 2
- IQFVPQOLBLOTPF-HKXUKFGYSA-L congo red Chemical compound [Na+].[Na+].C1=CC=CC2=C(N)C(/N=N/C3=CC=C(C=C3)C3=CC=C(C=C3)/N=N/C3=C(C4=CC=CC=C4C(=C3)S([O-])(=O)=O)N)=CC(S([O-])(=O)=O)=C21 IQFVPQOLBLOTPF-HKXUKFGYSA-L 0.000 description 54
- 238000006731 degradation reaction Methods 0.000 description 36
- 230000015556 catabolic process Effects 0.000 description 33
- 238000004042 decolorization Methods 0.000 description 32
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 24
- 239000010918 textile wastewater Substances 0.000 description 22
- 239000000975 dye Substances 0.000 description 20
- 240000004922 Vigna radiata Species 0.000 description 18
- 235000010721 Vigna radiata var radiata Nutrition 0.000 description 18
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 description 18
- 230000001580 bacterial effect Effects 0.000 description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- 238000001179 sorption measurement Methods 0.000 description 15
- 244000005700 microbiome Species 0.000 description 14
- 238000000034 method Methods 0.000 description 13
- 239000007788 liquid Substances 0.000 description 11
- 239000006228 supernatant Substances 0.000 description 11
- 239000000243 solution Substances 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 238000010790 dilution Methods 0.000 description 9
- 239000012895 dilution Substances 0.000 description 9
- 230000014759 maintenance of location Effects 0.000 description 9
- 239000000979 synthetic dye Substances 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 7
- 239000007857 degradation product Substances 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 239000002207 metabolite Substances 0.000 description 6
- 230000003287 optical effect Effects 0.000 description 6
- 241000193830 Bacillus <bacterium> Species 0.000 description 5
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 5
- 231100000674 Phytotoxicity Toxicity 0.000 description 5
- XRPLBRIHZGVJIC-UHFFFAOYSA-L chembl3182776 Chemical group [Na+].[Na+].NC1=CC(N)=CC=C1N=NC1=CC=C(C=2C=CC(=CC=2)N=NC=2C(=CC3=CC(=C(N=NC=4C=CC=CC=4)C(O)=C3C=2N)S([O-])(=O)=O)S([O-])(=O)=O)C=C1 XRPLBRIHZGVJIC-UHFFFAOYSA-L 0.000 description 5
- 239000000356 contaminant Substances 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 231100000331 toxic Toxicity 0.000 description 5
- 230000002588 toxic effect Effects 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 239000007621 bhi medium Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 4
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 102100022365 NAD(P)H dehydrogenase [quinone] 1 Human genes 0.000 description 3
- 241000589516 Pseudomonas Species 0.000 description 3
- 241000122971 Stenotrophomonas Species 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 125000000751 azo group Chemical group [*]N=N[*] 0.000 description 3
- 108010066657 azoreductase Proteins 0.000 description 3
- 238000010170 biological method Methods 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 238000005266 casting Methods 0.000 description 3
- 238000010835 comparative analysis Methods 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000002845 discoloration Methods 0.000 description 3
- 230000035784 germination Effects 0.000 description 3
- 238000002329 infrared spectrum Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 239000004753 textile Substances 0.000 description 3
- 231100000041 toxicology testing Toxicity 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108010029541 Laccase Proteins 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 150000004982 aromatic amines Chemical class 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- IJOOHPMOJXWVHK-UHFFFAOYSA-N chlorotrimethylsilane Chemical compound C[Si](C)(C)Cl IJOOHPMOJXWVHK-UHFFFAOYSA-N 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000543 intermediate Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000037353 metabolic pathway Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 238000001782 photodegradation Methods 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000002390 rotary evaporation Methods 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 238000004088 simulation Methods 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 238000002137 ultrasound extraction Methods 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 238000004065 wastewater treatment Methods 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000607528 Aeromonas hydrophila Species 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 241000186321 Cellulomonas Species 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 102000016680 Dioxygenases Human genes 0.000 description 1
- 108010028143 Dioxygenases Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010053759 Growth retardation Diseases 0.000 description 1
- 241000206596 Halomonas Species 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 108010059896 Manganese peroxidase Proteins 0.000 description 1
- 241000192041 Micrococcus Species 0.000 description 1
- 102000008109 Mixed Function Oxygenases Human genes 0.000 description 1
- 108010074633 Mixed Function Oxygenases Proteins 0.000 description 1
- XCOBLONWWXQEBS-KPKJPENVSA-N N,O-bis(trimethylsilyl)trifluoroacetamide Chemical compound C[Si](C)(C)O\C(C(F)(F)F)=N\[Si](C)(C)C XCOBLONWWXQEBS-KPKJPENVSA-N 0.000 description 1
- 229910019093 NaOCl Inorganic materials 0.000 description 1
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 150000001491 aromatic compounds Chemical class 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000012752 auxiliary agent Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000033558 biomineral tissue development Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 230000006652 catabolic pathway Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 239000011651 chromium Substances 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 125000000664 diazo group Chemical group [N-]=[N+]=[*] 0.000 description 1
- 108010005279 dichlorophenolindophenol reductase Proteins 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000005518 electrochemistry Effects 0.000 description 1
- 238000006056 electrooxidation reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000005189 flocculation Methods 0.000 description 1
- 238000005188 flotation Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- NYGZLYXAPMMJTE-UHFFFAOYSA-M metanil yellow Chemical group [Na+].[O-]S(=O)(=O)C1=CC=CC(N=NC=2C=CC(NC=3C=CC=CC=3)=CC=2)=C1 NYGZLYXAPMMJTE-UHFFFAOYSA-M 0.000 description 1
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 244000005706 microflora Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 238000006385 ozonation reaction Methods 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 238000011197 physicochemical method Methods 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 150000003254 radicals Chemical group 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- HFIYIRIMGZMCPC-YOLJWEMLSA-J remazole black-GR Chemical group [Na+].[Na+].[Na+].[Na+].[O-]S(=O)(=O)C1=CC2=CC(S([O-])(=O)=O)=C(\N=N\C=3C=CC(=CC=3)S(=O)(=O)CCOS([O-])(=O)=O)C(O)=C2C(N)=C1\N=N\C1=CC=C(S(=O)(=O)CCOS([O-])(=O)=O)C=C1 HFIYIRIMGZMCPC-YOLJWEMLSA-J 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 238000007142 ring opening reaction Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000024053 secondary metabolic process Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000010414 supernatant solution Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 239000005051 trimethylchlorosilane Substances 0.000 description 1
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 1
- 238000001429 visible spectrum Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/308—Dyes; Colorants; Fluorescent agents
-
- 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/30—Wastewater or sewage treatment systems using renewable energies
- Y02W10/37—Wastewater or sewage treatment systems using renewable energies using solar energy
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to an azo dye degrading bacterium and application thereof, wherein the azo dye degrading bacterium is enterococcus faecalis R1107 and is preserved in China Center for Type Culture Collection (CCTCC) M2022417. The azo dye degrading bacterium can also be applied to degrading azo dye wastewater, and the enterococcus faecalis R1107 can treat mixed STE containing azo dye, heavy metal and high salt under alkaline conditions, can effectively decolor the STE by 72.79%, and relieves the pH environment of high alkali. And not only has high decoloring efficiency, but also can effectively destroy the structure of azo dyes. LC-MS analysis results demonstrate that azo dyes can form low molecular weight, low toxicity compounds upon treatment with enterococcus faecalis R1107. The treatment of the azo dye by enterococcus faecalis R1107 reduces the toxicity obviously, namely, the azo dye is simply decolorized, and has the effects of biodegradation and detoxification.
Description
Technical Field
The invention relates to the field of dye wastewater treatment, in particular to azo dye degrading bacteria and application thereof.
Background
The waste water discharged from the textile industry containing azo dyes is highly coloured and can therefore be visually identified. Azo dyes have complex aromatic structures, are resistant to light, biological activity, ozone and other environmental conditions that are difficult to degrade. Thus, conventional wastewater treatment methods obviously fail to achieve the desired effect. In addition, anionic and nonionic azo dyes release toxic amines due to the reactive cleavage of azo groups. And the presence of heavy metals such as chromium, cobalt, nickel and copper (metallized dyes) in azo dye wastewater is also a concern for the environment.
At present, physical, chemical and biological methods are widely applied to the treatment of azo dye wastewater, and the treatment process is different according to the different dye molecular structures. Common physicochemical methods include adsorption, filtration, ozonation, UV/NaOCl, electrochemical oxidation, ultrasonic irradiation, UV/H2O2 conditioning, neutralization, photodegradation, electrochemistry, coagulation-flocculation, air flotation, precipitation, and the like. The methods release the harm caused by azo dye wastewater to a certain extent, but have limitations, such as toxic byproducts generated in the photodegradation process, simple adsorption and precipitation of high-concentration azo dye wastewater can not achieve the ideal effect, secondary degradation and complicated degradation process. The biodegradation method mainly comprises the technical methods of hydrolysis acidification, anaerobic biological method, aerobic biological method, biological adsorption, membrane separation, advanced oxidation method and the like. Among them, biological treatment has been proved to be an effective treatment method because of its low cost, high efficiency, simple operation, complete degradation, no secondary pollution, etc., and has received extensive attention. Conventional strategies for treating azo dye wastewater are mainly limited to physicochemical processes, which consume a great deal of energy and economic costs, and for these reasons, the current challenges focus on utilizing environmental bacteria capable of degrading azo dyes into small-molecule low-toxicity compounds.
To alleviate challenges such as difficulty in degrading azo dyes and sustained harmfulness, microbial bioremediation techniques have become an effective solution to colored water pollution. It is well known that microorganisms play a critical role in the mineralization of heterogeneous organic compounds. Early studies have demonstrated that dyes can be partially or completely biodegraded by fungi, algae, actinomycetes, plants and pure or mixed culture bacteria or enzymes thereof. Bacteria are ideal candidates for bioremediation as compared to other organisms, as they can be greatly adapted to the extreme environment of wastewater. In addition, bacterial cultures require shorter growth cycles and simple nutrition. Therefore, it is worth developing to utilize new bacteria in the environment for bioremediation. Bacteria known to be applied to azo dye wastewater degradation include bacillus, micrococcus, pseudomonas, bacillus, stenotrophomonas, halomonas, aeromonas hydrophila, bacillus anoxydans and the like.
It has been reported that a combination of three bacteria, stenotrophomonas acidophila, pseudomonas and cellulomonas, was developed for the degradation of monoazo dyes and that genetic analysis of each member demonstrated the individual biological versus dye degradation and catalytic potential. A degradation model is established by taking stenotrophomonas acidophilus, pseudomonas as a degradation dominant position and cellulose uniconas as a degradation auxiliary position. The scholars constructed a novel yeast population comprising Saccharomyces cerevisiae and Lactobacillus humanus producing manganese peroxidase and determined that its degradation products were not toxic aromatic amines. In addition, researchers have constructed a thermophilic microflora that effectively degrades azo dyes, and the synergistic effect of microorganisms such as anaerobic bacillus, clostridium, bacillus, etc. can secrete azo reductase and laccase to effectively decolorize azo dyes. Similarly, indian researchers isolated two strains of saline-alkaline bacteria secreted amylase from textile wastewater, which successfully decolorized the azo dye. Recently, a scholars can separate a strain of Inonomonas from textile wastewater, and can produce azo reductase, laccase and NADH-DCIP reductase, and has good degradation potential for the azo dye, namely, leishan red.
However, the current research is mostly focused on screening, characterization and degradation capability evaluation of azo dye degradation microorganisms, and the research improves our knowledge of azo dye degradation processes and to some extent of azo dye degradation processes. In order to improve the bioremediation efficiency of azo dye pollutants in the environment, besides screening azo dye degrading bacteria, the molecular mechanism of the strain for degrading azo dyes needs to be known. Previous studies have shown that the biodegradation of azo dyes is caused by a series of enzymes produced during their growth and metabolism, and that the bioremediation of the dyes can be achieved by bioadsorption and/or biodegradation. In the first case, the dye molecules bind to the surface of the organism; in the second case, the enzyme is responsible for converting complex dye molecules into simple compounds.
Bacterial biodegradation of azo dyes can occur under both aerobic and anaerobic conditions. Bacterial enzyme systems are capable of oxidative destruction of benzene rings, such as monooxygenases and dioxygenases. The degradation intermediate of azo dyes can be used as a nutrient source with the aid of other substrates and further mineralized. On the other hand, bacterial anaerobic degradation of azo dyes begins with the cleavage of azo bonds, followed by the formation of toxic intermediates, such as aromatic amines. Azo reductases are often reported to be involved in anaerobic degradation of azo dyes by bacteria. The target contaminant accepts electrons of the electron donor, thereby improving the complete degradation of the azo dye. In practical application, practical textile wastewater contains a large amount of heavy metals, salts and auxiliary agents besides synthetic dyes. The alkaline environment of textile wastewater limits the degradation efficiency of microorganisms. Microorganisms are generally considered less versatile in the presence of mixed contaminants. The harsh environment inhibits the growth of microorganisms and interrupts the biodegradation of the synthetic dye. At present, no microorganism which can still play a role in textile wastewater with harsh environment is found in the prior art.
Disclosure of Invention
Aiming at the defects, the invention provides an azo dye degrading bacterium and application thereof.
In order to solve the technical problems, the invention adopts the following technical scheme:
an azo dye degrading bacterium is enterococcus faecalis (Enterococcus faecalis) R1107, and is preserved in China center for type culture collection (CCTCC M2022417), wherein the preservation number is CCTCC M2022417, the preservation place is university of Wuhan, and the preservation date is 2022, 4 months and 14 days.
The application of the azo dye degrading bacteria in degrading azo dye wastewater comprises the following steps:
inoculating enterococcus faecalis R1107 with 5-15% inoculation amount into azo dye wastewater, and culturing at 30-40deg.C and 100-200rpm for more than 48 hr.
Further, the pH value of the azo dye wastewater is greater than 9.
Further, the inoculation amount of enterococcus faecalis R1107 is 10%.
The beneficial effects of the invention are as follows:
the invention separates and identifies a novel enterococcus faecalis R1107 from textile wastewater, researches the resistance and decoloration effect of the enterococcus faecalis R1107 on azo dyes, and verifies the effective degradation effect of the enterococcus faecalis R1107 on the azo dyes through Fourier transform infrared spectroscopy (FTIR) analysis. And the mixed STE containing azo dye, heavy metal and high salt can be treated under alkaline condition by utilizing the enterococcus faecalis R1107, and experiments prove that the enterococcus faecalis R1107 can effectively decolor the STE by 72.79 percent within 48 hours and relieve the pH environment of high alkali. And enterococcus faecalis R1107 not only has high decoloring efficiency, but also can effectively destroy the structures of azo dyes such as CR, RB5, DB38 and the like. LC-MS analysis revealed that azo dyes can form low molecular weight, low toxicity compounds upon treatment with enterococcus faecalis R1107. The phytotoxicity test result shows that the toxicity of the enterococcus faecalis R1107 treated with the azo dye is obviously reduced. In conclusion, the enterococcus faecalis R1107 has the effects of simple decolorization, biodegradation and detoxification on azo dyes.
The invention will now be described in detail with reference to the drawings and examples.
Drawings
FIG. 1 is a schematic diagram showing decolorization analysis of enterococcus faecalis R1107 on azo dye 0h (left 1) and 48h (right 2), wherein in FIG. 1 (A), the azo dye is Congo red, (B) the azo dye is reactive black 5, and (C) the azo dye is direct black 38;
FIG. 2 is a graph of time course analysis of E.faecalis R1107 decolorized with azo dyes of different concentrations; drawing and annotating: a: decolorization rate of Congo Red (CR) with different concentrations in 48h after E.faecalis R1107 treatment; b: the decolorization rate of active black five (RB 5) with different concentrations is treated by E.faecalis R1107 for 48 hours; c: the decolorization rate of direct black 38 (DB 38) with different concentrations is treated by E.faecalis R1107 for 48 hours;
FIG. 3 is a UV-visible spectrum scan of azo dye decolorization at different time intervals; drawing and annotating: a: congo Red (CR) at 0,2,6h decolorization; b: reactive black five (RB 5) at 0,2,6h decolorization; c: direct black 38 (DB 38) at 0,2,6h decolorization;
FIG. 4 is an infrared spectrum of untreated azo dye and enterococcus faecalis R1107 treated azo dye; drawing and annotating: a: congo Red (CR); b: reactive black five (RB 5); c: direct black 38 (DB 38);
FIG. 5 is a schematic of a phytotoxicity study of enterococcus faecalis R1107 treated and untreated azo dyes for 5 days; drawing and annotating: a: congo Red (CR); b: reactive black five (RB 5); c: direct black 38 (DB 38);
FIG. 6 is a graph of a colorimetric comparison analysis of the raw simulated textile wastewater (STE) with the simulated textile wastewater (T-STE) treated with E.faecalis R1107 for 48 hours.
Detailed Description
The principles and features of the present invention are described below with reference to the drawings, the examples are illustrated for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention.
1. Decolorization assay of enterococcus faecalis R1107
(1) Decolorization analysis of enterococcus faecalis R1107 on azo dyes with different concentrations
1. Marking the enterococcus faecalis R1107 glycerol bacteria preserved in a refrigerator at the temperature of minus 80 ℃ on an LB solid flat plate for activation, and culturing for 12 hours in a constant temperature incubator at the temperature of 37 ℃ until single bacterial colony grows out;
2. single colonies were picked up in BHI medium at 37℃and 150rpm to OD 600 0.8-1.0;
3. inoculating 10% of the strain to CR, RB5 and DB38 containing 50mg/L,100mg/L,200mg/L,500mg/L and 1000mg/L (azo dye detailed information is shown in BHI culture medium of Table 1, cultured at 37℃and 150rpm, and 1mL of sample was taken at 0h,6h,12h,24h,36h and 48 h;
4. centrifuging the sample at 12000rpm for 5min, removing thallus, collecting 200uL supernatant, and measuring OD in 96-well plate 497 ，OD 597 And OD (optical density) 585 Azo dyes without inoculating bacteria are used as blankControl [8] Each set of experiments was repeated three times;
5. CR, RB5 and DB38 decolorization rates at different times after enterococcus faecalis R1107 treatment were calculated as follows:
decoloration ratio (%) = [ (A) 0 -A)/A 0 ]X 100 formula (1)
In which A 0 : initial absorbance of azo dye, a: final absorbance of azo dye;
TABLE 1 azo dyes used in the present invention
(2) UV-visible detection of enterococcus faecalis R1107 treatment of different azo dyes
1. Marking the enterococcus faecalis R1107 glycerol bacteria preserved in a refrigerator at the temperature of minus 80 ℃ on an LB solid flat plate for activation, and culturing for 12 hours in a constant temperature incubator at the temperature of 37 ℃ until single bacterial colony grows out;
2. single colonies were picked up in BHI medium at 37℃and 150rpm to OD 600 0.8-1.0;
3. inoculating 10% of the strain into BHI culture medium containing 50mg/L CR, RB5 and DB38, culturing at 37deg.C and 150rpm, and taking 1mL samples at 0h,2h and 6h respectively;
4. centrifuging the sample at 12000rpm for 5min, removing thallus, collecting 200uL supernatant, and measuring OD on 96-well ELISA plate 200 -OD 800 Absorbance at the site, and each group of experiments was repeated three times with azo dye without inoculating the cells as a blank;
(3) Study of adsorption Rate of enterococcus faecalis R1107 on azo dyes with different concentrations
1. Streaking the preserved enterococcus faecalis R1107 glycerol bacteria on an LB solid plate for activation, and culturing for 12 hours in a constant temperature incubator at 37 ℃ until single colony is grown;
2. single colonies were picked up in BHI medium at 37℃and 150rpm to OD 600 0.8-1.0;
3. inoculating 10% of the strain into BHI culture medium containing 50mg/L,100mg/L,200mg/L,500mg/L,1000mg/L CR, RB5 and DB38, respectively, and culturing at 37deg.C and 150rpm for 48 h;
4. taking 1mL of sample, centrifuging at 12000rpm for 10min, thoroughly removing supernatant, collecting thallus, adding 70% methanol in equal volume, incubating at 37deg.C for 2 hr, eluting azo dye adsorbed on thallus [40] ；
Centrifugation at 5.12000 rpm for 10min, complete precipitation of the cells, taking 200uL supernatant and OD determination in 96 well plates 497 ，OD 597 And OD (optical density) 585 Taking azo dye with initial concentration as blank control;
6. the adsorption rate of enterococcus faecalis R1107 on different concentrations CR, RB5 and DB38 was calculated as follows:
adsorption rate (%) = [ a/a 0 ]X 100 formula (2)
In which A 0 : initial absorbance of azo dye, a: final absorbance of azo dye;
(4) FT-IR analysis of enterococcus faecalis R1107 treated azo dyes
1. Respectively inoculating 10% enterococcus faecalis R1107 to 50mL of BHI culture solution containing 50mg/L CR, RB5 and DB38, culturing at 37deg.C and 150rpm for 48 hr, and repeating each group of test three times with azo dye culture solution without inoculating thallus as blank control;
centrifuging at 2.12000 rpm for 10min, removing thallus, and collecting supernatant;
3.50 mL of culture supernatant, adding equal volume of ethyl acetate, and performing ultrasonic extraction for 10 min;
4. transferring the mixed liquid to a separating funnel, standing for layering, collecting a lower water phase, adding ethyl acetate with the same volume again for extraction, and repeating the steps for 3 times;
5. transferring ethyl acetate in the separating funnel to a new triangular flask;
6. anhydrous sodium sulfate (the bottom is fully paved, and water is removed for 1-2 h) is added;
7. the sample was freed from ethyl acetate by rotary evaporation under reduced pressure and dried overnight in a silica gel dryer;
8. ultrasonic dissolution with 2-3mL of methanol for 10min and filtration through 0.22 μm microporous filter membrane
9. Dripping 10uL of sample into dry potassium bromide powder, mixing, uniformly pressing on the surface of a cylinder by using a powder press to prepare a film, and taking the potassium bromide film as background data;
10. 8cm in 1min -1 Scanning for 90 times under resolution to obtain 400-4000cm -1 Is a spectrum of (2);
11. baseline and infrared spectral correction for penetration depth and frequency variation using Spectrum One software provided with the device and saving pictures [32] 。
2. LC-MS analysis of azo dyes treated by enterococcus faecalis R1107
1. Respectively inoculating 10% enterococcus faecalis R1107 to 50mL of BHI culture solution containing CR, RB5 and DB38 at 50mg/L, culturing at 37deg.C and 150rpm for 12 hr, taking azo dye culture solution without inoculating thallus as blank control and BHI culture solution as error control;
centrifuging at 2.12000 rpm for 10min, removing thallus, and collecting supernatant
3.50 mL of culture supernatant, adding equal volume of ethyl acetate, and performing ultrasonic extraction for 10 min;
4. transferring the mixed liquid to a separating funnel, standing for layering, collecting a lower water phase, adding ethyl acetate with the same volume again for extraction, and repeating the steps for 3 times;
5. transferring ethyl acetate in the separating funnel to a new triangular flask;
6. anhydrous sodium sulfate (the bottom is fully paved, and water is removed for 1-2 h) is added;
7. the sample was freed from ethyl acetate by rotary evaporation under reduced pressure and dried overnight in a vacuum silica gel dryer;
8. 100uL of pyridine and 100uL of derivatization reagent N, O-bis (trimethylsilyl) trifluoroacetamide (containing trimethylchlorosilane) are added into the sample, and the sample is incubated in a water bath at 60 ℃ for 10min and filtered through a 0.22 mu m microporous filter membrane for measurement;
9. the metabolite after azo dye decolorization was determined using an Agilent 1260-6224LC-MS TOF, C18 reverse phase column (4.6 mm. Times.250 mm, 5 um). Methanol and water were used as mobile phases with a flow rate of 1.0. 1.0 mL/min. The HPLC procedure used in the present invention was the same as in the previous study. The mass spectrum ranges from 100 to 500 m/z, and the measurement is carried out in a negative ion mode.
3. Toxicity study of enterococcus faecalis R1107 after treatment of azo dyes
1. The enterococcus faecalis R1107 single colony is picked up and cultured in BHI culture solution at 37 ℃ and 150rpm until OD 600 0.8-1.0;
2. inoculating the enterococcus faecalis R1107 bacterial liquid into BHI culture liquid with 10% of inoculation amount, respectively adding 50mg/L CR, RB5 and DB38, and culturing at 37 ℃ and 150rpm for 12h;
3. centrifuging the bacterial liquid at a low-temperature refrigerated centrifuge at 8000rpm and at 4 ℃ for 10min to remove bacterial bodies, and collecting a supernatant;
4. filtering and sterilizing the supernatant by a 0.22uM microporous filter membrane, and storing at 4 ℃;
5. placing 10 common mung beans selected randomly in a 90mm culture dish in an ultra-clean workbench, continuously adding 10mL of the supernatant solution for 5 days, taking water and BHI culture solution as blank control, placing the blank control in room temperature, observing the growth condition of the common mung beans, and repeating each group of experiments for three times;
6. taking out mung beans in the culture dish with forceps, washing with pure water, measuring the lengths of roots, stems and leaves of mung beans, and recording;
4. kinetic study of degradation of azo dyes by enterococcus faecalis R1107
1. The enterococcus faecalis R1107 single colony is picked up and cultured in BHI culture solution at 37 ℃ and 150rpm until OD 600 0.8-1.0;
2. taking the enterococcus faecalis R1107 bacterial liquid, inoculating the enterococcus faecalis R1107 bacterial liquid with 10% of inoculum size into a BHI culture liquid, respectively adding 50mg/L,100mg/L,200mg/L,500mg/L and 1000mg/L of RB5, and taking the non-inoculated bacterial liquid as a blank control;
3. taking 1mL of the sample in a 1.5mLEP tube every hour, and continuously sampling for 12 hours;
4. the sample was centrifuged at 12000rpm for 5min, and 200uL of supernatant was assayed in 96-well plates for OD 597 ；
5. The residual amount of enterococcus faecalis R1107 after treatment of azo dye RB5 in different time periods is calculated, and the formula is as follows:
residual amount (mg/L) = [ A/A ] 0 ]×50
In which A 0 : initial absorbance of azo dye, a: final absorbance of azo dye;
6.C t ＝-k 0 t+C 0 (n＝0)
C t ＝-C 0 e -k1t (n＝1)
C t -1 ＝k 2 +C 0 -1 (n＝2)
k 0 ,k 1 and k 2 Zero, primary and secondary rate constants, C 0 For the initial concentration of azo dye RB5, C t Is the concentration at a given time, and the dye concentration C is plotted separately t vs time t, lnC t vs time t and 1/C t -1 The zero order, first order and second order linear equations of vs time t are used for obtaining a correlation coefficient R 2 ；
1. The obtained data are used for determining a first order equation lnC according to dynamics by using origin software t ＝lnC 0 -kt fitting; wherein k: to remove rate constant, t: for the removal time of the contaminant azo dye, C 0 : for initial residual rate of contaminants, C t : is the residual rate of azo dye at different time points; contaminant removal half-life t 1/2=ln2/k
5. Study of enterococcus faecalis R1107 on simulation textile wastewater
1. Selecting enterococcus faecalis R1107 single colony, and culturing in BHI culture medium at 37deg.C and 150rpm for 6-8 hr;
2. the method is slightly modified on the basis of the literature, and the simulated textile wastewater is designed.
3. Inoculating enterococcus faecalis R1107 with 10% inoculation amount, culturing at 37deg.C in simulated textile wastewater at 150rpm for 48 hr, and repeating each group of experiments without inoculating enterococcus faecalis R1107 as blank control for three times;
4. centrifuging the bacterial liquid at 12000rpm for 10min, removing bacterial cells, and filtering the supernatant with a 0.22uM microporous filter membrane to obtain a sample;
5. the chromaticity of the simulated wastewater treated with enterococcus faecalis R1107 for 48 hours was determined by dilution factor method.
(1) The sample and the optical pure water are respectively taken into a colorimetric tube with a plug, the colorimetric tube with the plug is filled to a marked line, the colorimetric tube with the plug is placed on a white surface, the colorimetric tube with the plug and the surface are at a proper angle, so that the light rays are reflected by the bottom of the colorimetric tube with the plug upwards to pass through a liquid column, the sample and the optical pure water are compared, and the chromaticity and the tone presented by the sample are described, wherein the transparency is possibly included. Diluting until the solution is just indistinguishable from the optical pure water, and recording dilution factors;
(2) When the chromaticity of the sample is 50 times or more, the sample is pipetted into a volumetric flask, diluted with optically pure water to a standard line, and a large dilution ratio is obtained each time so that the dilution ratio is 50 times or less. When the chromaticity of the sample was 50 times or less, 25mL of the sample was taken in a cuvette with a plug, and diluted with optically pure water to a standard line, and the dilution factor was 2 each time. When the sample is diluted to a very low chromaticity, pouring a proper amount of sample from a colorimetric tube with a plug into a measuring cylinder, metering, diluting the sample to a marked line by using optical pure water, wherein the dilution multiple is less than 2 each time, and recording the values of the dilution multiple each time;
(3) Taking samples before and after STE degradation by bacteria to determine the pH value;
(4) The result shows that the dilution multiples of stepwise dilutions are multiplied by each other, and the multiplied product takes an integer value to represent the chromaticity of the sample, while describing the shade of the sample color in text, if transparency can be included.
Results and analysis
1. Decolorization analysis of enterococcus faecalis R1107 on azo dye
(1) Decolorization analysis of enterococcus faecalis R1107 on azo dyes with different concentrations
2. The invention screens out a strain which can degrade azo dye from textile wastewater, after separation and purification, to the Optimum of the Opt identification was performed by 16S rRNA sequencing, comparative analysis was performed at NCBI, identified as Enterococcus faecalis, and designated enterococcus faecalis R1107. The decolorization of three different azo dyes of enterococcus faecalis R1107 to CR, RB5 and DB38 was studied initially, and the decolorization efficiency of enterococcus faecalis R1107 to CR, RB5 and DB38 of 50mg/L,100mg/L,200mg/L,500mg/L,1000mg/L was monitored over 48 hours, with the visual effect shown in FIG. 1.
When the azo dye concentration was 50mg/L, the decolorization rates of CR, RB5 and DB38 were rapidly increased to 87.61%,83.08% and 69.77%, respectively, and the maximum decolorization rates of CR, RB5 and DB38 were 90.17%,96.82% and 81.95%, respectively, in 48 hours of treatment with enterococcus faecalis R1107. The high-concentration azo dye inhibits the decoloring potential of the enterococcus faecalis R1107 on the azo dye, and the decoloring rate of the enterococcus faecalis R1107 on the azo dye is continuously reduced along with the continuous increase of the concentration of the azo dye, but the high-concentration azo dye still has a good decoloring effect on high-concentration CR and RB 5. As shown in FIG. 2, when the concentration of azo dye was increased to 1000mg/L, the decolorization rate of enterococcus faecalis R1107 to CR and RB5 was rapidly increased over 6 hours to 47.51% and 65.07%, respectively, but the decolorization rate of DB38 was only 12%, and the decolorization rates of CR, RB5 and DB38 were increased to 65.57%, 72.64% and 23.39%, respectively, after 48 hours of treatment. The results show that the enterococcus faecalis R1107 has the best decolorizing effect on RB5, the influence of the dye concentration on the degradation efficiency is small, the CR is the next, the decolorizing efficiency is inhibited to a certain extent for the CR with high concentration, the concentration of DB38 is the key for limiting the decolorizing effect, and the decolorizing capability of the enterococcus faecalis R1107 is obviously inhibited by the excessive concentration. This is probably due to the difference in structural properties of CR, RB5 and DB38, and thus enterococcus faecalis R1107 has some difference in its decolorizing effect.
(2) UV-visible of enterococcus faecalis R1107 for treating different azo dyes
The supernatants of enterococcus faecalis R1107 treated with CR, RB5 and DB38 at different time periods were collected and subjected to UV-visible spectral scanning at 400-800nm, as compared to the 50mg/L CR, RB5 and DB38 without bacterial treatment.
The characteristic peaks of the azo dyes CR, RB5 and DB38 are respectively positioned at 497nm,597nm and 585nm, after being treated by enterococcus faecalis R1107 for 2 hours, the characteristic peaks of the azo dyes CR, RB5 and DB38 are respectively positioned at 497nm,597nm and 585nm, the significance of the characteristic peaks is reduced, and the peak area is obviously reduced. When the time of bacterial treatment was prolonged to 6 hours, the characteristic peaks at 497nm,597nm, and 585nm of CR, RB5, and DB38, respectively, disappeared nearly, and the visible spectrum peak pattern gradually tended to be smooth, and the result was shown in FIG. 3. This indicates that azo dyes CR, RB5 and DB38 after treatment with enterococcus faecalis R1107 had been stripped off in colour, the characteristic peaks disappeared and degradation occurred.
To date, about 60% of the synthetic dyes used in the textile industry are azo dyes, and a large part is not fixed on the product, which means that a large amount of azo dye is discharged into the waste water, and improving the decolorization performance of the synthetic dye at high concentration is a very interesting problem. Table 2 shows comparative analysis of dye discoloration at 1000 mg/L. In addition to DB38, enterococcus faecalis R1107 has better decolorizing potential for both CR and RB5 in a shorter period of time. As shown in the results of Table 2, most other microorganisms showed some degree of dye discoloration for high concentrations of synthetic dye, but the time for discoloration was longer compared to E.faecalis R1107 of the invention. The azo bonds formed in the azo dyes are stable and not readily degradable, CR and RB5 are defined as diazo dyes, and DB38 is defined as a triazo dye. Thus, the increase in azo bond in the synthetic dye may become a decrease in the decolorizing ability of enterococcus faecalis R1107 to azo dyes.
Table 2 comparative analysis of the decolorizing effect of microorganisms on high concentration (1000 mg/L) of dyes
(3) Adsorption analysis of enterococcus faecalis R1107 on azo dye
The dye may be adsorbed by microorganisms during the decolorization process. The invention evaluates the adsorption of enterococcus faecalis R1107 on azo dyes of different concentrations during decolorization, and the results are shown in Table 3. From this result, the microorganism has different adsorption effects on different concentrations of the azo dyes, and it is worth mentioning that among the three azo dyes used in the present invention, enterococcus faecalis R1107 has almost no adsorption effect on RB5, and the RB5 decolorization effect is not affected by the concentration thereof, which indicates that the key of RB5 decolorization is likely to be degradation, and enterococcus faecalis R1107 is also suitable for degradation of RB5 even under high concentration conditions.
For CR and DB38, when the dye concentration was increased to 1000mg/L, the decolorized adsorption of enterococcus faecalis R1107 on CR and DB38 was gradually increased, and the adsorption rates reached 42.59% and 13.72%, respectively. However, at low concentrations (50 mg/L) the adsorption rates were only 16.13% and 28.43%, indicating that degradation may be dominant and adsorption auxiliary for CR and DB38 decolorization.
TABLE 3 adsorption analysis of enterococcus faecalis R1107 on azo dyes of different concentrations
2. FT-IR analysis of enterococcus faecalis R1107 treated azo dyes
The invention researches the change of the functional group of azo dye after the azo dye is treated by enterococcus faecalis R1107 by adopting an infrared spectrometry. The FTIR spectra of azo dyes CR, RB5 and DB38 before and after treatment with enterococcus faecalis R1107 are shown in FIG. 4.
The infrared spectrum results show that enterococcus faecalis R1107 can effectively destroy the molecular structure of azo dyes. From the IR spectra of FIGS. 3A and 4A, untreated CR was found at 3400cm -1 Exhibits N-H stretching vibration at 2900cm -1 -3200cm –1 Exhibiting a C-H stretching vibration at 1600cm -1 The azo vibration absorption peak is at 1450cm -1 Where c=c stretch in the benzene ring; HO-SO 2 Stretching vibration of-OH at 1050cm- 1 -1200cm -1 Between 1000cm -1 The C-N stretching vibration is adopted, after the E.faecalis R1107 is treated, the transmittance of the indicator tape in CR is increased, and the absorption peak of azo vibration is weakened, which is probably that the azo bond of CR is destroyed.
FIGS. 3B and 4B show that N-H stretching is localized to 3450cm for untreated RB5 -1 O-H stretching is positioned at 3360cm -1 At 2930cm C-H -1 Stretching vibration at 1400cm -1 -1600 cm -1 Is a benzene ring with skeleton vibration of 1230cm -1 Is C-N stretch, -SO stretch positioned at 1100cm -1 Where RB5 was 1400cm after treatment with enterococcus faecalis R1107 -1 -1600cm -1 The stretching vibration at the position almost disappears,this suggests that RB5 has been decomposed by enterococcus faecalis R1107.
From FIGS. 3C and 4C, DB38 was at 3350cm -1 、2900cm- 1 、1680cm -1 And 1300cm -1 The peaks at these reflect the stretching vibration of-OH, stretching vibration of C-H, stretching vibration of c=c, and stretching vibration of C-N, respectively. After being treated by enterococcus faecalis R1107, the characteristic structure of DB38 is changed, in particular to the stretching vibration of benzene ring. This indicates that DB38 was biodegraded after treatment with enterococcus faecalis R1107.
3. LC-MS analysis of azo dyes treated by enterococcus faecalis R1107
With azo dyes CR, RB5 and DB38 as controls, possible degradation products of azo dyes after treatment with enterococcus faecalis R1107 were investigated and analyzed by LC-MS in the mode ESI TIC flag=175.0v. The detailed information is listed in table 4. The peak of CR degradation product appears at a retention time of 3.5min and at a retention time of 2.898min a peak molecular formula of 154m/z of C 12 H 10 The peak molecular formulas of 117m/z,119m/z and 120m/z appear at retention time of 3.065min as CH respectively 3 NaO 3 S，C 9 H 12 And C 8 H 11 N. The peak of RB5 degradation product appeared at a retention time of 3.5min. At a retention time of 2.973min, a peak of 179m/z occurs, with a molecular formula of C 6 H 5 NaO 3 S, the peak occurring at 3.423min retention time was 200m/z, and the molecular formula was inferred to be C 9 H 13 NO 2 S, S. The degradation products of DB38 appeared at retention times of 3.5min, peaks of 117m/z,134m/z and 151m/z appeared at retention times of 3.077 min, and molecular formulas were CH, respectively 3 NaO 3 S， C 9 H 13 N and C 8 H 13 N 3 . Peaks at retention times of 3.377min were 107m/z,134m/z, 174m/z and 200m/z, with molecular formulas C 6 H 8 N 2 ，C 2 H 5 NaO 2 S，C 4 H 10 NNaO 3 S and C 6 H 12 NNaO 3 S, S. Based on the detected metabolites, it is further demonstrated that azo dyes pass through enterococcus faecalisDegradation of R1107 occurs after treatment, and according to the results, possible degradation pathways of azo dyes are proposed, firstly asymmetric destruction of azo bonds, free radical ring opening, and formation of a series of different carboxylic acid byproducts of low molecular weight. These metabolites are greatly reduced in both toxicity and visual pollution compared to azo dyes. And simultaneously, the advantages of biodegradation are also reflected.
Table 4LC-MS data and chemical structures of CR, RB5 and DB38 degradation products
4. Toxicity analysis of enterococcus faecalis R1107 treated azo dyes
Plant toxicology studies were performed on azo dyes and their metabolites. The toxicity change of the azo dye before and after the enterococcus faecalis R1107 treatment on the common mung bean is analyzed and evaluated. It is well known that the toxicity of azo dyes threatens the health of the ecosystem, and the invention evaluates whether the toxicological risk of azo dyes after treatment with enterococcus faecalis R1107 can be effectively relieved by phytotoxicity study. In the present toxicology study, as shown in FIG. 5, the toxicity of azo dyes CR, RB5 and DB38 was evident, the growth and development of mung beans were remarkably suppressed compared with the blank group (water casting) after continuous 5d casting of 50mg/L azo dye, and the toxicity influence of azo dye on ordinary mung beans was remarkably reduced after casting of 5d azo dye treated with enterococcus faecalis R1107, and the growth suppression of azo dye on the same was alleviated. The root, stem and leaf lengths of common mung bean seeds under CR metabolite treatment were increased by 3.27-, 2.73-and 1.65-fold over those under CR treatment, respectively, as shown in panel A of FIGS. 3-5. The length of roots, stems and leaves of the common mung bean seeds treated by the RB5 metabolite is increased by 1.77-, 2.91-and 3.26 times respectively compared with that of the common mung bean seeds treated by the RB5 metabolite as shown in the B diagram in figures 3-5, and the length of the roots, stems and leaves of the common mung bean seeds treated by the DB38 metabolite is increased by 5.92-, 2.63-and 3.95 times respectively compared with that of the common mung bean seeds untreated. Meanwhile, the BHI culture medium also has an inhibition effect on the growth and development of common mung beans, but the azo dye is consumed by the degradation culture medium after the enterococcus faecalis R1107 is added, so that the germination of mung bean seeds is recovered.
As described in the previous studies, azo dyes are toxic aromatic compounds which are difficult to degrade, and in the present invention, the reduction of the length of roots, stems and leaves of mung beans under CR, RB5 and DB38 treatment indicates that azo dyes significantly inhibit the germination and growth of mung beans. In many cases, azo dyes can be degraded to low toxic compounds, even mineralized by microorganisms to carbon dioxide and water, and treatment of control group ingredients with BHI medium can be detrimental to common mung bean seeds, resulting in inhibited germination and growth of common mung bean seeds. In agreement with previous reports, the toxicity of azo dyes can be reduced by subjecting them to microbiological treatment. The degraded azo dye metabolite has lower phytotoxicity, which indicates that enterococcus faecalis R1107 has good potential in the aspect of biodegradation of textile dye.
5. Kinetic study of degradation of azo dye by enterococcus faecalis R1107
The degradation of RB5 by enterococcus faecalis R1107 is consistent with first order kinetics. As shown in Table 5, the decolorization time of R1107 enterococcus faecalis to RB5 was delayed as the concentration of azo dye RB5 was increased, increasing t1/2 of RB5 from 2.16h to 3.164h. Degradation rate also gradually slows down with increasing concentration of RB5, degradation rate k gradually decreases from 0.231 to 0.084, and the results conform to the first-order degradation kinetics. Meanwhile, the toxicity of RB5 is found to have a certain inhibition effect on the biological activity of enterococcus faecalis R1107, and particularly, the inhibition effect is more obvious when the concentration of RB5 is higher.
Table 5 E.faecalis R1107 analysis of the decolorization kinetics of RB5
6. Bioremediation of simulated textile wastewater
In order to further study the application potential of enterococcus faecalis R1107 in bioremediation, the simulation textile wastewater is subjected to bioremediation by adopting enterococcus faecalis R1107. Enterococcus faecalis R1107 has good decolorizing potential for STE, and the results are shown in FIG. 6.
After 48h treatment with enterococcus faecalis R1107, the color of STE was significantly reduced from 600℃to 163.26 ℃as shown in Table 5, and after bacterial treatment, the pH of STE was slightly reduced from 9.82 to 9.22.
The result shows that enterococcus faecalis R1107 has 72.79 percent of decoloration rate of STE and has a certain influence on the neutralization of alkaline wastewater. True textile waste water is generally composed of a number of different components, such as synthetic dyes, heavy metals and chemical adjuvants, and the high intensity of the textile waste water limits the penetration of sunlight into aquatic microorganisms, possibly leading to the photosynthesis turbulence of the aquatic ecosystem. The chemical structure of the synthetic dye is complex and difficult to treat, the microbial degradation efficiency of the mixed dye is generally reduced, and the co-stress of heavy metal and alkaline conditions in textile wastewater provides an extreme living environment for microorganisms, which is highly likely to cause apoptosis and death.
TABLE 6 decolorization analysis of enterococcus faecalis R1107 on simulated textile wastewater (STE)
The invention adopts enterococcus faecalis R1107 to improve the chromaticity of STE and adjust the pH thereof under alkaline environment. This suggests that enterococcus faecalis R1107's resistance to harmful chemicals and high salinity is beneficial for its further use in the bioremediation of textile wastewater.
Conclusion(s)
(1) Enterococcus faecalis R1107 has high decoloring efficiency to CR, RB5 and DB38 in 48 hours, the decoloring rates to CR, RB5 and DB38 of 50mg/L are 90.17%,96.82% and 81.95%, respectively, and the maximum decoloring rates to CR and RB5 still have certain decoloring effects when the concentration of azo dye reaches a high concentration of 1000mg/L reach 65.57% and 72.64%, respectively.
(2) FTIR analysis results show that enterococcus faecalis R1107 is able to effectively destroy the structure of azo dyes such as CR, RB5 and DB 38. LC-MS analysis revealed that azo dyes can form low molecular weight, low toxicity compounds upon treatment with enterococcus faecalis R1107. The phytotoxicity test result shows that the toxicity of the enterococcus faecalis R1107 treated with the azo dye is obviously reduced. In conclusion, the enterococcus faecalis R1107 has the effects of simple decolorization, degradation and detoxification on azo dyes.
(3) The invention proposes a possible mechanism for degrading azo dyes by enterococcus faecalis R1107 through the result of transcriptomic sequencing, and the sequencing result shows that the main metabolic pathway, secondary metabolism, transport system, amino acid metabolic pathway and signal transduction system of enterococcus faecalis R1107 are activated in the presence of RB 5. The synergistic effect of the enterococcus faecalis R1107 regulatory network promotes the degradation and detoxification of RB 5.
(4) In addition, the value of enterococcus faecalis R1107 in simulated textile wastewater was explored, and the result shows that enterococcus faecalis R1107 can treat mixed STE containing azo dye, heavy metal and high salt under alkaline conditions. Enterococcus faecalis R1107 may be able to decolorize STE within 48 hours effectively 72.79% and alleviate the high alkaline pH environment. The research result provides theoretical basis and potential application prospect for practical application of the textile wastewater bioremediation technology.
The foregoing is illustrative of the best mode of carrying out the invention, and is not presented in any detail as is known to those of ordinary skill in the art. The protection scope of the invention is defined by the claims, and any equivalent transformation based on the technical teaching of the invention is also within the protection scope of the invention.
Claims (4)
1. An azo dye degrading bacterium is characterized by being enterococcus faecalis (Enterococcus faecalis) R1107 and being preserved in China Center for Type Culture Collection (CCTCC) M2022417.
2. Use of the azo dye degrading bacterium according to claim 1 for degrading azo dye wastewater, characterized by comprising the steps of:
inoculating enterococcus faecalis R1107 with 5-15% inoculation amount into azo dye wastewater, and culturing at 30-40deg.C and 100-200rpm for more than 48 hr.
3. The use of an azo dye degrading bacterium according to claim 2 for degrading azo dye waste water, wherein the PH of the azo dye waste water is greater than 9.
4. The use of azo dye degrading bacteria according to claim 2 wherein the inoculation amount of enterococcus faecalis R1107 is 10%.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210443487 | 2022-04-25 | ||
| CN2022104434875 | 2022-04-25 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115369053A CN115369053A (en) | 2022-11-22 |
| CN115369053B true CN115369053B (en) | 2023-12-15 |
Family
ID=84062147
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210654954.9A Active CN115369053B (en) | 2022-04-25 | 2022-06-10 | Azo dye degrading bacterium and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115369053B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009066515A (en) * | 2007-09-13 | 2009-04-02 | Seiren Co Ltd | Method, device, and agent for decoloring dye-containing wastewater |
| CN108410751A (en) * | 2018-02-05 | 2018-08-17 | 温州大学 | A kind of Bacillus foecalis alkaligenes and its application in Degradation of Azo Dyes decoloration |
| CN110684687A (en) * | 2019-10-21 | 2020-01-14 | 汕头大学 | Enterococcus faecalis ST5 and application thereof in azo dye degradation |
| CN113173647A (en) * | 2021-04-26 | 2021-07-27 | 河北农业大学 | Bacillus amyloliquefaciens MN-13 and application thereof |
| CN113667615A (en) * | 2020-12-22 | 2021-11-19 | 南京工程学院 | Azo dye degrading bacterium and application thereof |
-
2022
- 2022-06-10 CN CN202210654954.9A patent/CN115369053B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009066515A (en) * | 2007-09-13 | 2009-04-02 | Seiren Co Ltd | Method, device, and agent for decoloring dye-containing wastewater |
| CN108410751A (en) * | 2018-02-05 | 2018-08-17 | 温州大学 | A kind of Bacillus foecalis alkaligenes and its application in Degradation of Azo Dyes decoloration |
| CN110684687A (en) * | 2019-10-21 | 2020-01-14 | 汕头大学 | Enterococcus faecalis ST5 and application thereof in azo dye degradation |
| CN113667615A (en) * | 2020-12-22 | 2021-11-19 | 南京工程学院 | Azo dye degrading bacterium and application thereof |
| CN113173647A (en) * | 2021-04-26 | 2021-07-27 | 河北农业大学 | Bacillus amyloliquefaciens MN-13 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115369053A (en) | 2022-11-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tan et al. | Aerobic decolorization and degradation of azo dyes by growing cells of a newly isolated yeast Candida tropicalis TL-F1 | |
| Saratale et al. | Decolorization and biodegradation of textile dye Navy blue HER by Trichosporon beigelii NCIM-3326 | |
| Saratale et al. | Enhanced decolorization and biodegradation of textile azo dye Scarlet R by using developed microbial consortium-GR | |
| Dhanve et al. | Biodegradation of diazo reactive dye Navy Blue HE2R (Reactive Blue 172) by an isolated Exiguobacterium sp. RD3 | |
| Dahiya et al. | Decolourisation of synthetic and spentwash melanoidins using the white-rot fungus Phanerochaete chrysosporium JAG-40 | |
| Bankole et al. | Biodegradation of a monochlorotriazine dye, cibacron brilliant red 3B-A in solid state fermentation by wood-rot fungal consortium, Daldinia concentrica and Xylaria polymorpha: Co-biomass decolorization of cibacron brilliant red 3B-A dye | |
| Mahmood et al. | Isolation and screening of azo dye decolorizing bacterial isolates from dye-contaminated textile wastewater. | |
| Ghosh et al. | Treatment of anaerobically digested distillery spentwash in a two-stage bioreactor using Pseudomonas putida and Aeromonas sp. | |
| Davies et al. | Plant and algal interference in bacterial beta-D-galactosidase and beta-D-glucuronidase assays | |
| Ayed et al. | Decolorization and phytotoxicity reduction of reactive blue 40 dye in real textile wastewater by active consortium: anaerobic/aerobic algal-bacterial-probiotic bioreactor | |
| CN103937726A (en) | Alga-lysing pseudomonas aeruginosa and application thereof | |
| Naik et al. | Enhanced degradation of melanoidin and caramel in biomethanated distillery spentwash by microorganisms isolated from mangroves | |
| Buvaneswari et al. | Original Research Article Bioremediation studies on sugar-mill effluent by selected fungal species | |
| CN101139140A (en) | Aerugo microcapsule algae degradation method using animalcule | |
| Karunya et al. | Development of microbial consortium CN-1 for the degradation of Mordant Black 17 | |
| Hanis et al. | Bacterial Degradation of Azo Dye Congo Red by Bacillus sp. | |
| Hamouda et al. | Removal of heavy metals and production of bioethanol by green alga Scenedesmus obliquus grown in different concentrations of wastewater | |
| Zhao et al. | Biodegradation and detoxification of the triphenylmethane dye coomassie brilliant blue by the extracellular enzymes from mycelia of Lactarius deliciosus | |
| US6613559B2 (en) | Simultaneous decolorization and detoxification of molasses spent wash using novel white rot-lignin-modifying fungus Flavodon flavus | |
| CN113173647B (en) | Bacillus amyloliquefaciens MN-13 and application thereof | |
| Saha et al. | Bioremediation of reactive orange 16 by industrial effluent-adapted bacterial consortium VITPBC6: process optimization using response surface methodology (RSM), enzyme kinetics, pathway elucidation, and detoxification | |
| Dewi et al. | Optimization of the conditions for the decolorization of batik wastewater by Aspergillus sp. 3 | |
| El-Sheekh et al. | Biodegradation of some dyes by the cyanobacteria species Pseudoanabaena sp. and Microcystis aeruginosa Kützing. | |
| CN115369053B (en) | Azo dye degrading bacterium and application thereof | |
| CN102965298B (en) | Lysine bacillus and method for degrading MC-LR by the same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |