CN111018131A - Method for degrading sulfamethoxazole - Google Patents

Method for degrading sulfamethoxazole Download PDF

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CN111018131A
CN111018131A CN201911293348.3A CN201911293348A CN111018131A CN 111018131 A CN111018131 A CN 111018131A CN 201911293348 A CN201911293348 A CN 201911293348A CN 111018131 A CN111018131 A CN 111018131A
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sulfamethoxazole
sterilized
culture medium
degrading
medium
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陈银广
李悦
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Tongji University
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Tongji University
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    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/28Anaerobic digestion processes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/30Organic compounds
    • C02F2101/34Organic compounds containing oxygen
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/30Organic compounds
    • C02F2101/38Organic compounds containing nitrogen
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/30Organic compounds
    • C02F2101/40Organic compounds containing sulfur

Abstract

The invention discloses a method for degrading sulfamethoxazole, which degrades sulfamethoxazole by the synergistic effect of paracoccus denitrificans P. The invention reduces the accumulation of intermediate products under the interaction of the microorganisms and finally realizes the anaerobic denitrification.

Description

Method for degrading sulfamethoxazole
Technical Field
The invention belongs to the technical field of environmental protection, and particularly relates to a method for degrading sulfamethoxazole.
Background
Sulfonamide antibiotics are widely used in the fields of human medical treatment, animal disease prevention and treatment, livestock breeding and the like. Research has shown that about 85% of the medicinal sulfonamides antibiotics can not be absorbed by human body or animals, so they can be discharged out of the body, and finally cause environmental pollution. Antibiotics not only cause chemical pollution of the environment, but also induce microorganisms in the environment to mutate, generate resistance genes and spread. Microorganisms carrying resistance genes can enter human bodies through food chains, so that the efficacy of drugs can be influenced after the human bodies generate resistance to certain drugs, and the health of the human bodies is harmed, so that the removal of antibiotics in sewage, underground water and soil is paid more and more attention.
The main methods for degrading antibiotics at present are a flocculation precipitation method, a chemical oxidation method, an adsorption method and the like. While the general physical method essentially transfers the antibiotic from the environment without degrading it, the chemical oxidation method produces many undefined intermediates, which are even more toxic than the antibiotic originally.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, it is an object of the present invention to provide a method for degrading sulfamethoxazole by synergistic action of paracoccus denitrificans p.
In order to achieve the above objects and other objects, the present invention provides a method for degrading sulfamethoxazole by the synergistic effect of paracoccus denitrificans and shewanella.
In one embodiment, the process for degrading sulfamethoxazole by the synergistic effect of paracoccus denitrificans and shewanella comprises at least the following steps:
providing a reactor;
adding a culture medium, mother liquor containing sulfamethoxazole and trace elements into the reactor, and sterilizing to obtain a sterilized culture medium system;
preparing paracoccus denitrificans suspension and Shewanella suspension;
adding the paracoccus denitrifican suspension, the shewanella suspension and sterilized calcium chloride into the sterilized culture medium system to degrade sulfamethoxazole.
In one embodiment, Paracoccus denitrificans and Shewanella are cultured under aerobic conditions and concentrated to obtain a concentrated suspension of Paracoccus denitrificans and Shewanella.
In one embodiment, the reactor is a sterile anaerobic bottle.
In one embodiment, the components of the medium include a carbon source and an inorganic salt. The concentration of the carbon source is 0 g/L-5 g/L.
In one embodiment, the carbon source is any one or combination of glucose, sodium acetate, sodium lactate, and sodium succinate, wherein the glucose is sterilized in a separate container.
In one embodiment, the inorganic salt comprises 0-7.2 g/L potassium nitrate, 0-0.5 g/L ammonium chloride, 0-0.1 g/L magnesium sulfate, 0.5-4.65 g/L disodium hydrogen phosphate, and 1-2.44 g/L potassium dihydrogen phosphate.
In one embodiment, the composition of the trace elements includes disodium edetate, ferrous sulfate heptahydrate, manganese chloride, sodium molybdate, copper chloride crystals, and zinc chloride.
In one embodiment, the initial OD of Paracoccus denitrificans6000.07 to 0.31, the initial OD of the Shewanella6000.22 to 0.42.
In one embodiment, the pH of the sterilized medium system is 6.8-7.4.
In one embodiment, the temperature condition in the process of degrading sulfamethoxazole is 20-50 ℃.
In one embodiment, in the step of adding the culture medium, the mother liquor containing sulfamethoxazole and the trace elements into the reactor and performing the sterilization treatment, the sterilization treatment is sterilized by high-pressure steam. And (2) sterilizing the calcium chloride solution and/or the glucose, and sterilizing the calcium chloride solution and the glucose by using high-pressure steam, wherein the calcium chloride solution and the glucose are independently sterilized, but the temperature is 110-115 ℃ when the glucose is sterilized. The paracoccus denitrificans is Paracoccus denitificans (ATCC13543) and the Shewallenanoneidensis is Shewallenanoneidensis MR-1(ATCC 700550).
In the invention, the Shewallenanoneidensis MR-1 is added in the process of degrading sulfamethoxazole by Paracoccus denitrificans, and the anaerobic denitrification is realized through the mutual cooperation of the two bacteria, so that the degradation efficiency is improved, and the degradation amount is ensured to be at a higher level. Meanwhile, the Shewanella is a common strain in underground water, and compared with methods for improving the degradation of sulfamethoxazole by photolysis treatment, hydrolysis treatment, chemical oxidation treatment and the like, the method saves cost and does not produce intermediate products with higher toxicity.
Drawings
Fig. 1 is a schematic flow chart of a method according to an embodiment of the present invention.
Detailed Description
The following description of the embodiments of the present invention is provided by way of specific examples, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the disclosure herein. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention. It is to be noted that the features in the following embodiments and examples may be combined with each other without conflict. It is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the respective manufacturers.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs and the description of the present invention, and any methods, apparatuses, and materials similar or equivalent to those described in the examples of the present invention may be used to practice the present invention.
According to the invention, the Shewallenanoneidensis MR-1 is added in the process of degrading sulfamethoxazole by Paracoccus denitrificans, so that anaerobic denitrification is realized through the mutual cooperation of the two bacteria, the degradation efficiency is improved, and the degradation amount is ensured to be at a higher level.
Referring to fig. 1, the present invention provides a method for degrading sulfamethoxazole, wherein the process for degrading sulfamethoxazole by the synergistic effect of paracoccus denitrificans and shewanella comprises at least the following steps:
s1, providing a reactor;
s2, adding a culture medium, mother liquor containing sulfamethoxazole and trace elements into the reactor, and sterilizing to obtain a sterilized culture medium system;
s3, preparing paracoccus denitrificans suspension and Shewanella suspension;
s4, adding the paracoccus denitrificans suspension, the Shewanella suspension and sterilized calcium chloride into the sterilized culture medium system to degrade sulfamethoxazole.
Specifically, in step S1, the reactor is a sterile anaerobic bottle.
Specifically, in the step S2, the components of the medium include a carbon source and inorganic salts. The concentration of the carbon source is 0 g/L-5 g/L. The carbon source is any one or combination of glucose, sodium acetate, sodium lactate and sodium succinate, wherein the glucose is sterilized in a separate container. The inorganic salt comprises 0-7.2 g/L potassium nitrate, 0-0.5 g/L ammonium chloride, 0-0.1 g/L magnesium sulfate, 0.5-4.65 g/L disodium hydrogen phosphate and 1-2.44 g/L potassium dihydrogen phosphate. The trace elements comprise disodium ethylene diamine tetraacetate, ferrous sulfate heptahydrate, manganese chloride, sodium molybdate, copper chloride crystals and zinc chloride. The sterilization treatment is carried out by autoclaving. Adjusting the pH value of the system to 6.8-7.4 by using sodium hydroxide NaOH and hydrochloric acid HCl, and then sealing the reactor by using a sealing film of a sterile culture container for sterilizing for 20-30 minutes.
Specifically, in step S3, the initial OD of Paracoccus denitrificans6000.07 to 0.31, the initial OD of the Shewanella6000.22 to 0.42. The temperature condition in the sulfamethoxazole degradation process is 20-50 ℃, and the temperature in the sulfamethoxazole degradation process is critical to the reaction and can influence the activity of microbial bacteria. The concentration of nitrate in the culture medium is 0-1000 mg/L. Respectively culturing the cells in sterilized 100-200mL LB culture medium under aerobic conditionNitrification microorganisms Paracoccus denitificas and Shewallenaneonidensis MR-1 to OD600For 2 and 1.2, the cultured denitrifying microorganisms Paracoccus denitificans and Shewallenanoneidensis MR-1 were washed three times with phosphate buffer and concentrated to 40-50 mL.
Specifically, in the step S4, 20-25g/L of calcium chloride CaCl is prepared2Sterilizing with high pressure steam for 20-30 min, and sterilizing at 110-115 deg.C for 15-20 min to prepare 500g/L glucose solution, wherein the calcium chloride solution and glucose solution are sterilized separately. The purpose of the calcium chloride addition is to accelerate the reaction, and if not, to make it almost non-reactive. Adding 0-1.2 mL of sterilized glucose into 100-200mL of sterilized culture medium system, adding 80-100 μ l of sterilized CaCl2 solution, and finally respectively adding 0.1-5 mL of concentrated paradenitrococcus suspension and 0.5-5 mL of Shewanella suspension to degrade sulfamethoxazole, wherein the temperature condition in the process of degrading sulfamethoxazole is 20-50 ℃. The denitrifying microorganism paracoccus denitrificans Paracoccus denitrificans is singly inoculated into a degradation culture medium such as LB culture medium as a control experiment, and is placed on a shaking bed at the temperature of 20-50 ℃ to carry out the degradation experiment of sulfamethoxazole. The control group is arranged to be beneficial to enhancing the reliability of the experimental result.
The concentration of sulfamethoxazole in the invention is detected by high performance liquid chromatography, and the specific detection process and conditions are as follows: 1.5ml of the sample was centrifuged at 4000-. The instrument model is Agilent1220 type High Performance Liquid Chromatography (HPLC), chromatographic column: agilent Eclipse Plus C18(5 μm, 2.1X 150 mm); mobile phase 0.1% aqueous formic acid: 0.1% formic acid acetonitrile 70:30 (volume ratio v/v); the detection wavelength is 275 nm; flow rate 0.6 ml/min; the sample volume was 20. mu.l, and the column temperature was 25 ℃.
In one embodiment, a culture medium, 10-12 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system A, the pH of the system is adjusted to 7.4 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system A is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the culture mediumThe components comprise inorganic salt and carbon source, wherein the inorganic salt comprises 2.16 g-2.5/LKNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: : na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600The denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 cultured in advance were washed three times with PBS and concentrated to 40mL for 2 and 1.2. Add 600. mu.l sterilized glucose to 100mL sterilized Medium System A to make the glucose concentration 3g/L, add 100. mu.l sterilized CaCl2 solution, and finally add 3mL concentrated P.Denitrificans suspension to the medium to make OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.22 to 0.24. Placing on a constant temperature shaking bed at 30 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 62.7%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 10-12 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system B, the pH of the system is adjusted to 7.4 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system B is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes:Na2-EDTA (7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2o (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively6002 and 1.2; the previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100mL of sterilized culture medium system B to ensure that the concentration of the glucose is 2.8-3 g/L, then adding 100 mu L of sterilized CaCl2 solution, and finally respectively adding 3mL of concentrated P.Denitrificans suspension into the degradation culture medium to ensure that OD is achieved6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.40 to 0.42. Placing on a constant temperature shaking bed at 30 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 72.4%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 10-12 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system C, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system C is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively6002 and 1.2; the previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100mL of sterilized culture medium system C to ensure that the concentration of the glucose is 2.8-3 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 30 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 69.2%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 10-12 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system D, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system D is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600For 2 and 1.2, previously cultured denitrification was microminiaturizedOrganisms P.dentificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100mL of sterilized culture medium system D to enable the glucose concentration to be 2.8-3 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 20 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The results show that the degradation rate of sulfamethoxazole is 17.4%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 8-10 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system E, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system E is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100ml of sterilized culture medium system E to enable the glucose concentration to be 3-3.2 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally adding the degradation culture medium into the culture medium respectivelyOD 3mL of concentrated P.Denitrificans suspension6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 82.6%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 10-12 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system F, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system F is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100ml of sterilized culture medium system F to ensure that the concentration of the glucose is 3-3.2 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 50 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The results show thatThe degradation rate of sulfamethoxazole is 3.3%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 4.5-5.5 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system G, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is adopted for sealing, and the system is sterilized at the temperature of 121 ℃ for 20 minutes to obtain the sterilized culture medium system G. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/LKNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100ml of sterilized culture medium system G to ensure that the concentration of the glucose is 2.5-3G/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The results show that the degradation rate of sulfamethoxazole is 75.7%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 19-21 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system H, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system H is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salts and a carbon source, wherein the inorganic salts comprise 2.16 to E2.5g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively6002 and 1.2; the previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. To 100ml of the sterilized medium system H, 600. mu.l of sterilized glucose was added so that the glucose concentration became 3g/L, and 100. mu.l of sterilized CaCl2 solution was added. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 93.2%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 19-21 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system I, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system I is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4,3.85~4g/L CH3COOH. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively6002 and 1.2; the previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. To 100ml of sterilized medium system I, 100. mu.l of sterilized CaCl2 solution was added. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The results show that the degradation rate of sulfamethoxazole is 8.5%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 19-21 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system J, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system J is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO44.85-5 mL/L sodium lactate solution. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes. Separately culturing the denitrifying Micronitrites in advance in aerobic conditions with sterilized, for example, 100mL LB MediumOrganisms P.dentificans and S.oneidenesis MR-1 to OD600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensisMR-1 were washed three times with PBS and concentrated to 40 mL. To 100ml of sterilized medium system J was added 100. mu.l of sterilized CaCl2 solution. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The results show that the degradation rate of sulfamethoxazole is 37.1%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 19-21 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system K, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system K is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively6002 and 1.2; the previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 1.2mL of sterilized glucose into 100mL of sterilized culture medium system K to enable the glucose concentration to be 4.5-5 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Most preferably3mL of each concentrated suspension of P.dentificans was added to the degradation medium to make OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 87.7%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 19-21 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system L, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system L is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the medium comprises inorganic salts and carbon source, wherein the inorganic salts comprise 0.5g/L NH4Cl,0.1g/L MgSO4,4.65g/L Na2HPO4,2.44g/LKH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD, respectively, are cultured in advance under aerobic conditions with sterilized, for example 100mLLB medium600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. To 100ml of the sterilized medium system T, 600. mu.l of sterilized glucose was added so that the glucose concentration became 3g/L, and 100. mu.l of sterilized CaCl2 solution was added. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The results show that sulfamethoxazoleThe degradation rate of (2) was 34.0%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 19-21 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system M, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system M is sterilized at 121 ℃ for 20 minutes to obtain the sterilized culture medium system M. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 3.5-3.7 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100ml of sterilized culture medium system M to ensure that the concentration of the glucose is 3-3.2 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 94.2%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 19-21 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture mediumAdjusting the pH value of the system to 6.8 by NaOH and HCl, sealing by using a sealing film of an aseptic culture container, and sterilizing at 121 ℃ for 20 minutes to obtain a sterilized culture medium system N. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 7-7.1 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. To 100ml of the sterilized medium system N, 600. mu.l of sterilized glucose was added so that the glucose concentration became 3g/L, and 100. mu.l of sterilized CaCl2 solution was added. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 14.3%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 10-12 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system O, the pH of the system is adjusted to 7.4 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture container is sterilized at 121 ℃ for 20 minutes to obtain the sterilized culture medium system O. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100ml of sterilized culture medium system O to ensure that the concentration of the glucose is 3-3.2 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally, 0.1mL of each of the concentrated P.Denitrificans suspensions was added to the degradation medium to obtain OD6000.07-0.09, and adding 1mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.21 to 0.23. Placing on a constant temperature shaking bed at 30 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The results show that the degradation rate of sulfamethoxazole is 18.6%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 10-12 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system P, the pH of the system is adjusted to 7.4 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system P is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 2.16-2.5 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Denitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are cultured in advance under aerobic conditions with sterilized, for example, 100mL of LB medium, respectively600The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 600 mu L of sterilized glucose into 100ml of sterilized culture medium system P to ensure that the concentration of the glucose is 3-3.2 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally, 5mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.31-0.33, and adding 1mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.42 to 0.44. Placing on a constant temperature shaking bed at 30 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 68.9%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
In one embodiment, a culture medium, 19-21 mg/L sulfamethoxazole and 1mL of trace elements are added into a sterile anaerobic bottle to form a culture medium system Q, the pH of the system is adjusted to 6.8 by NaOH and HCl, a sealing film of a sterile culture container is used for sealing, and the sterile culture medium system Q is obtained after sterilization at the temperature of 121 ℃ for 20 minutes. Wherein the components of the culture medium comprise inorganic salt and a carbon source, wherein the inorganic salt comprises 3.5-3.7 g/L KNO3,0.5g/L NH4Cl,0.07~0.1g/L MgSO4,2~4.65g/L Na2HPO4,1~2.44g/L KH2PO4. The trace element composition (g/L) includes: na (Na)2-EDTA(7.30)、FeSO4·7H2O(2.50)、MnCl2·4H2O(0.02)、Na2MoO4·2H2O(0.242)、CuCl2·2H2O (0.135) and ZnCl2(0.34). Then 20g/LCaCl was prepared2Sterilized at 121 ℃ for 20 minutes and 500g/L glucose at 115 ℃ for 15 minutes. Preparation ofDenitrifying microorganisms P. Denitrificans and S. oneidensis MR-1 to OD are first cultured in sterilized, for example, 100mL LB medium under aerobic conditions, respectively600Are 2 and 1.2. The previously cultured denitrifying microorganisms P.dentitificans and S.oneidensis MR-1 were washed three times with PBS and concentrated to 40 mL. Adding 1.2mL of sterilized glucose into 100mL of sterilized culture medium system Q to ensure that the concentration of the glucose is 3-5 g/L, and then adding 100 mu L of sterilized CaCl2 solution. Finally, 3mL each of the concentrated P.Denitrificans suspensions was added to the degradation medium to OD6000.18-0.20, and adding 0.5mL of concentrated S.oneidensis MR-1 suspension to obtain the final OD6000.32 to 0.34. Placing on a constant temperature shaking bed at 35 ℃, carrying out anaerobic culture for 7 days, and carrying out a degradation experiment of sulfamethoxazole. The result shows that the degradation rate of sulfamethoxazole is 96.8%. Denitrifying microorganisms P. dentrificans were inoculated alone into degradation medium as a control.
TABLE 1 comparison of data for some examples of the invention
According to the data analysis in table 1 and the examples, it is found that when the concentration of paracoccus denitrificans is determined, the additional Shewanella promotes the degradation of sulfamethoxazole SMX, but even if the concentration of Shewanella is changed, the degradation efficiency of SMX is not greatly influenced. The degradation efficiency of SMX increases with decreasing pH, which is maximal at pH 6.8. Under acidic conditions, the activity of relevant enzymes for degrading SMX is increased, so that the degradation efficiency of SMX can be increased. At low concentrations of SMX, it has less inhibition of the microorganism, but at relatively high concentrations of SMX, the rate of the enzymatic reaction can be increased, but too high a concentration of SMX inhibits the activity of the microorganism. Therefore, in order to realize the enzymatic reaction rate which is low in inhibition and can be improved by achieving high substrate concentration, a balance needs to be found, and therefore the 20-22 mg/LSMX degradation effect is good. Sodium acetate, glucose, sodium lactate and sodium succinate are all common carbon sources, glucose is the most suitable carbon source for paracoccus denitrificans, and sodium lactate is the most suitable carbon source for Shewanella. Sodium succinate is beneficial for the denitrification process of the mixed bacteria system, but is basically utilized for SMX metabolism. When sodium lactate is used as a carbon source, the SMX degradation process is partially performed, because it is the best carbon source for sodium lactate and can also be used by paracoccus denitrificans, but Shewanella is the dominant one, and thus the effect is not particularly desirable. Glucose is the most suitable carbon source for paracoccus denitrificans, and sodium lactate generated by metabolism of the glucose under anaerobic conditions can be utilized by Shewanella, so that the optimal state of the two bacteria is further achieved, and the reaction rate of SMX degradation can be greatly accelerated. The degradation rate of SMX increases with increasing glucose concentration, and when the glucose concentration reaches a certain amount, increasing the glucose concentration does not increase the degradation rate of SMX. The effect of nitrogen source concentration on the degradation of SMX by microorganisms is critical to the concentration of carbon source. Although a high concentration of nitrogen source can promote the respiration of the microorganism and synthesize more cell material, under the condition of a certain carbon source concentration, the excessive nitrogen source can consume all carbon sources, so that no redundant carbon source is supplied to the microorganism for growth and metabolism. The degradation efficiency is greatly influenced by temperature, and when the temperature is lower than 35 ℃, the degradation efficiency is greatly reduced because the temperature influences the activities of two microorganisms, and in the embodiment of the invention, for example, when the temperature is 50 ℃, the degradation rate is only about 3.3 percent. The calcium chloride of the present invention is intended to accelerate the reaction rate and is one of inorganic salts because it is easily deposited in the medium if sterilized together with the medium, and affects its effect. Glucose sterilized alone is also prevented from settling in the medium, and glucose is also a carbon source, because if sterilized together with the medium, glucose also easily settles in the medium, affecting its effect.
According to the invention, the Shewallenanoneidensis MR-1 is added in the process of degrading sulfamethoxazole by Paracoccus denitrificans, so that anaerobic denitrification is realized through the mutual cooperation of the two bacteria, the degradation efficiency is improved, and the degradation amount is ensured to be at a higher level. Meanwhile, the Shewanella is a common strain in underground water, and compared with methods for improving the degradation of sulfamethoxazole by photolysis treatment, hydrolysis treatment, chemical oxidation treatment and the like, the method saves cost and does not produce intermediate products with higher toxicity. In addition, the method of the invention is beneficial to human health and improves the environment.
Furthermore, it is to be understood that one or more method steps mentioned in the present invention does not exclude that other method steps may also be present before or after the combined steps or that other method steps may also be inserted between these explicitly mentioned steps, unless otherwise indicated; it should also be understood that unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention which may be practiced, nor is it intended to limit the relative changes or modifications to the scope of the invention which may be practiced without materially changing the technical details.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.

Claims (10)

1. A method for degrading sulfamethoxazole, which is characterized in that sulfamethoxazole is degraded by the synergistic effect of paracoccus denitrificans and Shewanella.
2. The method for degrading sulfamethoxazole according to claim 1, wherein the process of degrading sulfamethoxazole by the synergistic effect of paracoccus denitrificans and shewanella comprises at least the following steps:
providing a reactor;
adding a culture medium, mother liquor containing sulfamethoxazole and trace elements into the reactor, and sterilizing to obtain a sterilized culture medium system;
preparing paracoccus denitrificans suspension and Shewanella suspension;
adding the paracoccus denitrifican suspension, the shewanella suspension and sterilized calcium chloride into the sterilized culture medium system to degrade sulfamethoxazole.
3. The method for degrading sulfamethoxazole according to claim 2, wherein the reactor is a sterile anaerobic bottle.
4. The method for degrading sulfamethoxazole according to claim 3, wherein the components of the culture medium include a carbon source and inorganic salts.
5. The method for degrading sulfamethoxazole according to claim 4, wherein the carbon source is any one or more of glucose, sodium acetate, sodium lactate and sodium succinate, and the glucose is sterilized in a separate container.
6. The method for degrading sulfamethoxazole according to claim 4, wherein the composition of the inorganic salt comprises 0-7.2 g/L of potassium nitrate, 0-0.5 g/L of ammonium chloride, 0-0.1 g/L of magnesium sulfate, 0.5-4.65 g/L of disodium hydrogen phosphate and 1-2.44 g/L of potassium dihydrogen phosphate.
7. The method for degrading sulfamethoxazole according to claim 2, wherein the composition of the trace elements comprises disodium ethylenediamine tetraacetic acid, ferrous sulfate heptahydrate, manganese chloride, sodium molybdate, copper chloride crystals and zinc chloride.
8. The method for degrading sulfamethoxazole according to claim 2, wherein the initial OD of paracoccus denitrificans is6000.07 to 0.31, the initial OD of the Shewanella6000.22 to 0.42.
9. The method for degrading sulfamethoxazole according to claim 2, wherein the pH of the sterilized culture medium system is 6.8-7.4.
10. The method for degrading sulfamethoxazole according to claim 2, wherein the temperature condition in the process of degrading sulfamethoxazole is 20-50 ℃.
CN201911293348.3A 2019-12-16 2019-12-16 Method for degrading sulfamethoxazole Pending CN111018131A (en)

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