CN114573118A - Method for removing o-dichlorobenzene, m-dichlorobenzene and p-dichlorobenzene from water by using mucor circinelloides and application of method - Google Patents
Method for removing o-dichlorobenzene, m-dichlorobenzene and p-dichlorobenzene from water by using mucor circinelloides and application of method Download PDFInfo
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Abstract
The invention discloses a method for removing o-dichlorobenzene, m-dichlorobenzene and p-dichlorobenzene in water by using mucor circinelloides and application thereof. The preservation name of the mucor circinelloides FNZJ3-2-2 isMucor circinelloidesFNZJ3-2-2, deposited in the China general microbiological culture Collection center, with the deposition address of No. 3 Xilu No. 1 Beijing, Chaoyang, the collection number of CGMCC No.9684, and the collection date of 2014, 09 and 19 days; the eutrophic wastewater is an inorganic salt liquid culture medium containing three mixed dichlorobenzene of ortho-dichlorobenzene, meta-dichlorobenzene and para-dichlorobenzene; the inorganic salt liquid culture medium consists of K2HPO4•3H2O；NaH2PO4；(NH4）2SO4(ii) a pH6.0-7.0. The single-factor test proves that the Mucor circinelloides FNZJ3-2-2 living bacteria have strong degradation effect on the o-dichlorobenzene, the m-dichlorobenzene and the three mixed dichlorobenzene in the eutrophic wastewater.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a method for removing o-dichlorobenzene, m-dichlorobenzene and p-dichlorobenzene in water by using mucor circinelloides and application thereof.
Background
Compared with the traditional chemical technology and physical technology for removing organic pollutants, the biodegradation has more economic and environmental feasibility, and becomes an environment-friendly and energy-saving sewage (waste water) and waste gas treatment technology with great prospect. Research shows that various microorganisms show good biodegradation effects on various single organic pollutants or combined organic pollutants, and the microorganisms can degrade the organic pollutants by using a unique carbon source or being metabolized together with other organic matters. The microorganism has strong adaptability and mutability, can mutate all organic pollutants to form corresponding degrading microorganisms, and has very high repairing application prospect in degrading the organic pollutants by using the microorganisms.
Persistent Organic Pollutants (POPs) such as organochlorine pesticides (OCPs), polybromodiphenyl ethers (PBDEs), halogenated hydrocarbons, Polycyclic Aromatic Hydrocarbons (PAHs), etc., are of great concern for environmental pollution. These contaminants pose a significant threat to human health even at low concentrations. To date, there have been many reports on the use of different biomasses for the removal of organic contaminants from aqueous solutions. Three kinds of dichlorobenzene, namely o-dichlorobenzene, m-dichlorobenzene and p-dichlorobenzene, are selected as typical organochlorine pollutants, and the degradation capability of mucor circinelloides FNZJ3-2-2 is researched. The mechanisms of dichlorobenzene biodegradation are generally oxidative and reductive dechlorination catalyzed by enzymes. Under aerobic conditions, the degradation reaction of chlorobenzene compounds basically follows a mechanism of ring opening and dechlorination. Firstly, under the action of dioxygenase, inserting oxygen atom into aromatic ring to form cyclic chlorodiol, then removing two hydrogen atoms by catalase and converting them into chlorocatechol. The reductive dechlorination refers to a process that a chlorobenzene compound removes a chlorine substituent while obtaining electrons and releases a chlorine anion, and the dechlorination of the chlorobenzene compound is a reductive dechlorination process under an anaerobic or anoxic condition.
Under the closed culture condition, the concentration of the residual volatile dichlorobenzene in the gas above the culture bottle is measured by headspace gas chromatography, and the degradation efficiency of dichlorobenzene is calculated according to the prepared dichlorobenzene volatilization standard curve.
CN104830702A discloses Mucor circinelloides FNZJ3-2-2 and application thereof in nickel electroplating bath solution for aluminum profiles. The aluminum profile nickel electroplating bath solution is a high-nickel solution of sulfuric acid and boric acid, and contains complex and strong-acid heavy metal components. The patent does not disclose that the strain can be applied to removal of dichlorobenzene, and the biodegradation, tolerance and resistance thereof are not yet clarified.
Disclosure of Invention
The invention mainly solves the technical problem of providing the application of the mucor circinelloides FNZJ3-2-2 in removing the o-dichlorobenzene, the m-dichlorobenzene and the p-dichlorobenzene in the eutrophic wastewater, and single-factor tests prove that the living thalli of the mucor circinelloides FNZJ3-2-2 have strong degradation effects on the o-dichlorobenzene, the m-dichlorobenzene and the p-dichlorobenzene in the eutrophic wastewater.
In order to solve the technical problems, the invention adopts a technical scheme that: application of mucor circinelloides FNZJ3-2-2 in removing o-, m-and p-dichlorobenzene in eutrophic wastewater, wherein the mucor circinelloides FNZJ3-2-2 under the deposit name ofMucor circinelloidesFNZJ3-2-2, preserved in China general microbiological culture Collection center, with the preservation address of No. 3 Xilu No. 1 Beijing, Chaoyang, the preservation number of CGMCC No.9684, and the preservation date of 2014, 09 months and 19 days;
the eutrophic wastewater is an inorganic salt liquid culture medium containing three mixed dichlorobenzene of ortho-dichlorobenzene, meta-dichlorobenzene and para-dichlorobenzene;
the inorganic salt liquid culture medium consists of K2HPO4·3H2O；NaH2PO4；(NH4)2SO4； pH6.0-7.0。
Further, the composition of the inorganic salt liquid culture medium is K2HPO4·3H2O，10-15g/L； NaH2PO4，1-5g/L；(NH4)2SO4，1-5g/L；pH6.0-7.0。
Further, the composition of the inorganic salt liquid culture medium is K2HPO4·3H2O, 13.755g/L (or K)2HPO4，10.5g/L)；NaH2PO4，4.5g/L；(NH4)2SO4，1g/L；pH7.0。
Furthermore, the adding ratio of the o-dichlorobenzene, the m-dichlorobenzene and the p-dichlorobenzene in the eutrophic wastewater is 1:1: 1.
Further, the application is as follows: adding a proper amount of living bacteria of mucor circinelloides FNZJ3-2-2 into the eutrophic wastewater containing o-, m-and p-dichlorobenzene so that the initial bacteria content in the reaction liquid after inoculation is 2-10 multiplied by 108Culturing the strain per mL for 24-72 h under the conditions of degradation temperature of 28-36 ℃ and rotation speed of 180 r/min.
More specifically, the nutrient-rich wastewater containing o-, m-and p-dichlorobenzene has an initial concentration of 25-125 mg/L and an initial degradation pH of 6.0-8.0.
Furthermore, the initial bacteria content of the added Mucor circinelloides FNZJ3-2-2 living bacteria is 6X 108one/mL.
More specifically, the nutrient-rich wastewater refers to a liquid culture medium containing 50mg/L of each of ortho-dichlorobenzene, meta-dichlorobenzene and para-dichlorobenzene.
Still further, the initial degradation pH was 7.0.
More particularly, the degradation temperature is 32 ℃.
In some embodiments, the invention features the use of Mucor circinelloides for the removal of one or more of ortho-, meta-, or para-dichlorobenzene from water.
In some preferred embodiments, the Mucor circinelloides accession number is CGMCC No. 9684.
In some embodiments, the present invention features a method for removing one or more of ortho-, meta-, or para-dichlorobenzene from water using Mucor circinelloides, comprising the steps of:
inoculating Mucor circinelloides to water containing o-dichlorobenzene, m-dichlorobenzene or p-dichlorobenzene with the pH of 6.0-8.0, and culturing at the temperature of 28-36 ℃ under shaking, preferably 29-33 ℃, and more preferably 31-33 ℃.
In some preferred embodiments, the shaking culture is a shaking culture, preferably with a shaking rotation speed of 120-250 rpm.
In some preferred embodiments, the shaking culture time is 24-72 h.
In some preferred embodiments, the pH of the water containing o-dichlorobenzene, m-dichlorobenzene, or p-dichlorobenzene is 5.8-6.2.
In some preferred embodiments, the pH of the water containing o-dichlorobenzene, m-dichlorobenzene, or p-dichlorobenzene is 6.3 to 6.7.
In some preferred embodiments, the pH of the water containing o-dichlorobenzene, m-dichlorobenzene, or p-dichlorobenzene is 6.3 to 7.3.
In some preferred embodiments, the content of the initial bacteria of the Mucor circinelloides after inoculation is 2 to 10X 108one/mL, preferably 10X 108one/mL.
In some preferred embodiments, the concentration of o-dichlorobenzene, m-dichlorobenzene, or p-dichlorobenzene in water is 25-125 mg/L, preferably 20-30 mg/L or 80-110 mg/L.
In some preferred embodiments, the weight ratio of o-dichlorobenzene, m-dichlorobenzene, or p-dichlorobenzene to water is (1-5): (1-5): (1-5), preferably (1-2): (1-2): (1-2).
In some preferred embodiments, the water further comprises an inorganic salt, preferably the inorganic salt comprises K2HPO4、NaH2PO4And (NH)4)2SO4At least one of (1). Said K2HPO4May be derived from K2HPO4·3H2O or K without water of crystallization2HPO4。
The research finds that the water contains K2HPO4、NaH2PO4And (NH)4)2SO4It is necessary to degrade ortho-, meta-or para-dichlorobenzene. In addition, the mucor circinelloides can well grow and degrade o-dichlorobenzene, m-dichlorobenzene or p-dichlorobenzene without adding extra magnesium, manganese and iron salts.
In some preferred embodiments, the K is2HPO4The concentration of the water in the water is 10-11 g/L.
In some preferred embodiments, the NaH2PO4The concentration of the water in the water is 4-5 g/L.
In some preferred embodiments, the K is2HPO4The concentration of the water in the water is 0.5-1.5 g/L.
In some preferred embodiments, characterized in that the mucor circinelloides is in log phase.
In some preferred embodiments, the Mucor circinelloides is cultured in LB culture solution to a late logarithmic phase, centrifuged at 13000r/min for 10min at 4 ℃, sterilized water is added for 3 times, and the cells are kept as viable bacteria and preserved at 4 ℃ for later use.
The invention has the beneficial effects that:
single factor tests prove that the Mucor circinelloides FNZJ3-2-2 living bacteria have strong degradation effects on o-dichlorobenzene, m-dichlorobenzene and three kinds of mixed dichlorobenzene in the eutrophic wastewater.
Drawings
FIG. 1 is a graph showing the dichlorobenzene removal rate of the live bacteria FNZJ3-2-2 of the invention at different initial degradation pH values;
FIG. 2 is a graph showing the dichlorobenzene removal rate of the live bacteria FNZJ3-2-2 of the present invention at different degradation temperatures;
FIG. 3 is a graph showing the dichlorobenzene removal rate of the viable bacteria FNZJ3-2-2 of the present invention at different initial dichlorobenzene concentrations;
FIG. 4 is a graph showing the dichlorobenzene removal rate of the live bacterial strain FNZJ3-2-2 of the present invention at different initial bacterial contents.
Detailed Description
The following detailed description of the preferred embodiments of the present invention, taken in conjunction with the accompanying drawings, will make the advantages and features of the present invention more comprehensible to those skilled in the art, and will thus provide a clear and concise definition of the scope of the present invention.
Preservation of strains
Mucor circinelloides FNZJ3-2-2 is preserved in China general microbiological culture Collection center of China general microbiological culture Collection management Committee at 19.09.2014, with the preservation address of No. 3 Siro No. 1 of Beijing market, North Chen of the sunward region and the preservation number of CGMCC No. 9684. The separation and identification method of the strain is described in patent ZL 201510293835.5.
Example 1: an application of mucor circinelloides FNZJ3-2-2 in removing the mixed o-, m-and p-dichlorobenzene in the eutrophic waste water,
1. preparation of cells
1000mL of Mucor circinelloides LB culture solution in logarithmic phase is taken, centrifuged for 10min at 4 ℃ and 13000r/min, added with sterile water and repeatedly centrifuged for 3 times, and constant-weight wet cells are taken as live bacteria and stored at 4 ℃ for later use.
2. Biodegradation one-factor test of FNZJ3-2-2 live bacteria
Taking a proper amount of live bacteria of Mucor circinelloides FNZJ3-2-2 to ensure that the density of bacteria in the system is 2-10 multiplied by 108Per mL, add enriched dichlorobenzene containing three mixtures of o-, m-and p-dichlorobenzeneCulturing the nutrient wastewater for 24-72 h under the conditions that the pH value is 6.0-8.0, the degradation temperature is 28-36 ℃, the concentration of the three o-dichlorobenzene, m-dichlorobenzene and p-dichlorobenzene is 25-125 mg/L (the three o-dichlorobenzene and the three p-dichlorobenzene are 1:1:1 and are respectively 25-125 mg/L) and the rotation speed is 180r/min, and exploring a single-factor suitable condition for removing mixed dichlorobenzene pollutants from the FNZJ3-2-2 living bacteria in the nutrient-rich wastewater. The preset central values of the single factors are respectively as follows: pH 7.0, culture temperature 32 deg.C, initial bacteria content 6 × 108o/mL, o-m-p-three dichlorobenzene 1:1:1, 50mg/L each. The concentration of the residual volatile dichlorobenzene in the upper layer gas in the culture flask is measured by gas chromatography, and the biological removal rate of the paradichlorobenzene in the FNZJ3-2-2 strain is calculated.
(1) Under the condition that the central value of other single factors is not changed, the pH is changed to be 6.0, 6.5, 7.0, 7.5 and 8.0 respectively, and the result is shown in figure 1;
(2) under the condition that the central value of other single factors is not changed, the culture temperature is changed to be 28 ℃, 30 ℃, 32 ℃, 34 ℃ and 36 ℃ respectively, and the result is shown in figure 2;
(3) under the condition that the central value of other single factors is not changed, the concentrations of ortho-dichlorobenzene, meta-dichlorobenzene and para-dichlorobenzene (1: 1: 1) are respectively changed to be 25mg/L, 50mg/L, 70mg/L, 75mg/L, 100mg/L and 125mg/L, and the result is shown in figure 3;
(4) under the condition of other single-factor central value being unchanged, the initial bacteria concentration is changed to be 2.0 multiplied by 10 respectively8cell/mL, 4.0X 1082/mL, 6.0X 1082/mL, 8.0X 108one/mL, 10.0X 108The results are shown in FIG. 4.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all equivalent structural changes made by using the contents of the present specification and the drawings, or applied directly or indirectly to other related technical fields, are included in the scope of the present invention.
Claims (10)
1. Use of Mucor circinelloides for the removal of one or more of ortho-, meta-or para-dichlorobenzene from water.
2. Use according to claim 1, characterized in that: the preservation number of the mucor circinelloides is CGMCC No. 9684.
3. A method for removing one or more of ortho-dichlorobenzene, meta-dichlorobenzene or para-dichlorobenzene from water by using Mucor circinelloides through Mucor circinelloides, which is characterized by comprising the following steps:
water containing o-dichlorobenzene, m-dichlorobenzene or p-dichlorobenzene, the pH value of the water is 6.0-8.0, mucor circinelloides is inoculated, and the water is subjected to shaking culture at the temperature of 28-36 ℃, preferably 29-33 ℃, and more preferably 31-33 ℃.
4. The method according to claim 3, wherein the shaking culture is shaking culture, preferably with a shaking rotation speed of 120-250 rpm;
and or, the shaking culture time is 24-72 h.
5. The method according to claim 3 or 4, wherein the pH of the water containing o-dichlorobenzene, m-dichlorobenzene or p-dichlorobenzene is 5.8-6.2;
and/or the pH of the water containing o-dichlorobenzene, m-dichlorobenzene or p-dichlorobenzene is 6.3-6.7;
and/or the pH value of the water containing o-dichlorobenzene, m-dichlorobenzene or p-dichlorobenzene is 6.3-7.3.
6. The method according to any one of claims 3 to 5, wherein the initial content of Mucor circinelloides after inoculation is 2 to 10X 108one/mL, preferably 10X 108one/mL.
7. The method according to any one of claims 3 to 6, wherein the concentration of o-dichlorobenzene, m-dichlorobenzene, or p-dichlorobenzene in water is 25 to 125mg/L, preferably 20 to 30mg/L or 80 to 110 mg/L;
and or, the weight ratio of the o-dichlorobenzene, the m-dichlorobenzene or the p-dichlorobenzene in the water is (1-5): (1-5): (1-5), preferably (1-2): (1-2): (1-2).
8. The method according to any one of claims 3 to 7, wherein the water further comprises an inorganic salt, preferably wherein the inorganic salt comprises K2HPO4、NaH2PO4And (NH)4)2SO4At least one of (1).
9. The method of any one of claims 3-8, wherein K is2HPO4The concentration of the water in the water is 10-11 g/L;
and or, the NaH2PO4The concentration of the water in the water is 4-5 g/L;
and or, said K2HPO4The concentration of the water in the water is 0.5-1.5 g/L.
10. The method of any one of claims 3 to 9, wherein the mucor circinelloides is in log phase;
preferably, the Mucor circinelloides is subjected to LB culture solution culture to a post-logarithmic phase, centrifuged at 13000r/min for 10min at 4 ℃, added with sterile water and repeatedly centrifuged for 3 times, and constant-weight wet cells are used as live bacteria and stored at 4 ℃ for later use.
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| SU1652335A1 (en) * | 1989-06-08 | 1991-05-30 | Институт Биохимии И Физиологии Микроорганизмов Ан Ссср | Actinomyces strain streptomyces rochei, carrying out complete degradation of 2,4,6-trichlorophenol, or 2,4-dichlorophenol, or 2,6- dichlorophenol, or 2-chlorophenol |
| JP2010252673A (en) * | 2009-04-23 | 2010-11-11 | National Institute For Agro-Environmental Science | Cyclic dienic compound-degrading microorganism and cyclic dienic compound-degrading agent, apparatus for cleaning polluted environment, and method for degrading cyclic dienic compound and method for cleaning pollution substance |
| CN105483026A (en) * | 2015-12-31 | 2016-04-13 | 广东省生态环境与土壤研究所 | Mucor circinelloides H3 and application thereof in pyrene degradation |
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2022
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|---|---|---|---|---|
| SU1652335A1 (en) * | 1989-06-08 | 1991-05-30 | Институт Биохимии И Физиологии Микроорганизмов Ан Ссср | Actinomyces strain streptomyces rochei, carrying out complete degradation of 2,4,6-trichlorophenol, or 2,4-dichlorophenol, or 2,6- dichlorophenol, or 2-chlorophenol |
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