CN116496931B - Enterobacter cloacae, preparation method thereof, culture medium and application - Google Patents

Enterobacter cloacae, preparation method thereof, culture medium and application Download PDF

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CN116496931B
CN116496931B CN202211738828.8A CN202211738828A CN116496931B CN 116496931 B CN116496931 B CN 116496931B CN 202211738828 A CN202211738828 A CN 202211738828A CN 116496931 B CN116496931 B CN 116496931B
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enterobacter cloacae
degradation
trichlorophenol
anaerobic
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CN116496931A (en
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王燕
仝泽方
马英辉
卢美欢
李利军
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Microbiology Institute Of Shaanxi
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F11/00—Treatment of sludge; Devices therefor
    • C02F11/02—Biological treatment
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/02—Separating microorganisms from their culture media
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/04—Preserving or maintaining viable microorganisms
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/36—Adaptation or attenuation of cells
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/045—Culture media therefor
    • 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
    • C02F2101/34—Organic compounds containing oxygen
    • C02F2101/345—Phenols
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Abstract

The invention discloses enterobacter cloacae and a preparation method, a culture medium and application thereof, wherein the preservation number of the enterobacter cloacae in China center for type culture collection is CCTCC NO: m20221208 is a facultative enterobacter cloacae strain which is separated and screened from the oil field sludge of the elm in Shaanxi province by the inventor, has high growth speed under aerobic condition and can rapidly degrade 2,4, 6-trichlorophenol under anaerobic condition. Compared with the existing chlorophenol biodegradation strain, the enterobacter cloacae prepared by the invention has better culture economic characteristics and anaerobic degradation characteristics, and the degradation rate of the 2,4, 6-trichlorophenol can reach more than 80 percent.

Description

Enterobacter cloacae, preparation method thereof, culture medium and application
Technical Field
The invention belongs to the technical field of environmental microbial degradation, and particularly relates to enterobacter cloacae and a preparation method and application thereof.
Background
Chlorophenols are a class of toxic and environmentally persistent organic pollutants that are primarily derived from improper emissions during use, and are widely used in the wood preservation, metal rust protection, preservative, pesticide, herbicide industries. At present, the existence of chlorophenols is detected in the environments of soil, groundwater, wastes, sludge and the like, and the chlorophenols are easy to accumulate in biological circles through food chains because of certain water solubility, so that the chlorophenols have posed a great threat to the environment and human health. The european union, united states, japan, china, and many other countries have listed chlorophenols as preferential contaminants.
Conventional methods for removing chlorophenols contaminants include physical methods (adsorption method, air stripping, membrane separation), chemical methods (wet oxidation method, supercritical oxidation method, high-frequency ultrasonic method, high-voltage pulse discharge, tiO2 membrane photocatalysis) and microbiological methods. The microbial method is more important because of the characteristics of less secondary pollution, economy and the like. In nature, microorganism resources are abundant, corresponding degrading bacteria are easy to find in places where organic matters are concentrated, the existing strains capable of degrading chlorophenols are mainly concentrated in corynebacteria, pseudomonas, acetic acid bacteria, bacillus and the like, aerobic degradation of low chlorophenols is mainly used, and reports of anaerobic degradation of the occasional chlorophenols are also researches on the aspects of concentrated community structure diversity. In general, the microbial degradation of polychlorophenone (more than trichlorophenol) requires dechlorination under anaerobic conditions to form low-chlorophenol or phenol, and then ring-opening degradation.
Anaerobic dechlorination refers to the use of anaerobic microorganisms under anaerobic or anoxic conditions as an organic carbon source or hydrogen (H) 2 ) And taking 2,4,6-TCP as a respiratory chain end electron acceptor as an electron donor, and removing chlorine substituents on benzene rings and generating energy for the growth of microorganisms. In this process, 2,4,6-TCP is gradually converted into organic products (phenol) with lower chlorine content (mono-chlorophenol, dichlorophenol) and even complete dechlorination, the toxicity is greatly reduced, the degradability is improved, and the organic products are more easily decomposed by microorganisms so as to realize complete mineralization of pollutants (into carbon dioxide and water). Anaerobic dechlorination is therefore the first step for polychlorophenone and is the critical rate limiting step for the complete degradation of polychlorophenone.
Microbial agents capable of performing anaerobic dechlorination degradation on polychlorinated phenols in the prior reports are mainly concentrated in Dehalococcoides genus and Desulfobacillus genus, and mainly comprise anaerobic bacteria. However, the anaerobic bacteria have the disadvantages of high culture conditions, complicated operation procedures, high maintenance cost and low degradation efficiency. Therefore, the microbial resource with the dechlorination function is further excavated, the functional diversity of dechlorination microbes is enriched, and the method has great significance for the treatment of actual chlorophenol pollution.
Disclosure of Invention
In order to solve the problems, the invention aims to explore and develop a microbial agent capable of degrading 2,4, 6-trichlorophenol and culturing at low cost and enrich microbial resources with dechlorination function.
Based on the above purpose, on one hand, the invention utilizes 2,4, 6-trichlorophenol as the sole carbon source to carry out gradual gradient stress domestication, combines the nutrient medium to carry out anaerobic-aerobic alternate culture to separate and screen a facultative strain which can grow fast aerobically and degrade 2,4, 6-trichlorophenol fast from sludge, and the preserving information of the facultative strain is as follows:
strain name: OST-12;
latin name: enterobacter cloacae
Preservation unit: china center for type culture Collection;
preservation number: cctccc NO: m20221208;
preservation time: 2022, 8 and 1.
Furthermore, the 16S rRNA nucleotide sequence of the enterobacter cloacae provided by the invention is SEQ ID No. 1.
In another aspect, the invention also provides a screening method of enterobacter cloacae, which comprises the following steps:
(1) Preparing a culture medium;
(4) Sludge activation and gradual domestication aerobic-anaerobic alternate culture;
(5) Separating and screening degradation strains;
the sludge activation and gradual domestication aerobic-anaerobic alternate culture is to mix activated sludge with an activation culture medium, fill sterile nitrogen, seal and stand to prepare an activated sludge culture;
the activated sludge culture is alternately subjected to anaerobic culture in a domestication degradation culture medium and aerobic culture in a liquid nutrition culture medium;
the concentration of 2,4, 6-trichlorophenol in the domesticated degradation culture medium is gradually increased when the culture is alternately performed for the same activated sludge culture, namely aerobic culture and anaerobic culture.
Further, the concentration of the 2,4, 6-trichlorophenol in the domesticated degradation culture medium is gradually increased from 10mg/L to 100mg/L according to the domestication requirement during alternate culture.
Further, the separation and screening of the degradation strain is to dilute and spread the culture solution obtained by gradual domestication aerobic-anaerobic alternate culture on a degradation flat plate containing 100mg/L of 2,4, 6-trichlorophenol degradation screening flat plate culture medium, culture in an anaerobic box, after bacterial colonies grow out, pick single colonies on the flat plate on a nutrient agar culture medium flat plate, carry out aerobic culture, respectively pick the colonies into a liquid domestication degradation culture medium containing 100mg/L of 2,4, 6-trichlorophenol, fill sterile nitrogen, stand in the anaerobic box for culture, centrifugally remove supernatant, pour the supernatant into a liquid domestication degradation culture medium containing 100mg/L of 2,4, 6-trichlorophenol which is fresh for sterilization, and continuously carry out liquid phase measurement on the degradation rate of the final 2,4, 6-trichlorophenol after anaerobic culture.
In a third aspect, the present invention also provides a domesticated degradation medium, the components of the domesticated degradation medium comprising: ammonium chloride, sodium thiosulfate, potassium dihydrogen phosphate, sodium pyruvate, calcium chloride dihydrate, magnesium chloride dihydrate, cysteine, trace element liquid, vitamin liquid, distilled water and 2,4, 6-trichlorophenol.
Preferably, the domestication degradation culture medium comprises the following components in percentage by mass;
the adding ratio of the trace element liquid to the vitamin liquid to the distilled water is 1:4:200 by volume ratio.
In a fourth aspect, the invention also provides a degradation screening plate culture medium, which is obtained by adding agar according to 1.5-2% in a domestication culture medium basis.
In a fifth aspect, the present invention also provides an activation medium comprising the components of: sodium lactate, sodium bicarbonate, sodium chloride, magnesium chloride hexahydrate, calcium chloride dihydrate, yeast extract, ammonium chloride, potassium dihydrogen phosphate, trace element liquid and distilled water.
Furthermore, the invention also provides application of the enterobacter cloacae in degrading 2,4, 6-trichlorophenol, and the degradation rate can reach more than 80% through degradation test.
By combining the technical scheme and the embodiment, the enterobacter cloacae OST-12 provided by the invention has at least the following beneficial effects or advantages:
1. the enterobacter cloacae OST-12 provided by the invention belongs to facultative anaerobism, has low nutrition requirement, can grow rapidly in a common aerobic culture medium, can accumulate higher biomass, can efficiently degrade trichlorophenol in an anaerobic environment, enriches the diversity of 2,4, 6-trichlorophenol degrading microorganism families, and is beneficial to low-cost culture in application.
2. According to the enterobacter cloacae OST-12 provided by the invention, the degradation rate of the 2,4, 6-trichlorophenol can reach more than 80% through the degradation rate test.
Drawings
FIG. 1 is a graph showing the comparison of the degradation rates of 2,4, 6-trichlorophenol by 8 bacteria selected in example 1.
FIG. 2 shows the morphology and microscopic morphology of the Enterobacter cloacae OST-12 plates.
FIG. 3 is a tree of the Enterobacter cloacae OST-12.
FIG. 4 shows the carbon source utilization characteristics of Enterobacter cloacae OST-12.
FIG. 5 shows the degradation profile of Enterobacter cloacae OST-12 versus 2,4, 6-trichlorophenol.
Detailed Description
The following describes the technical aspects of the present invention with reference to examples, but the present invention is not limited to the following examples.
The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products available on the market without the manufacturer's attention.
Example 1 preparation of Medium
Sludge activation medium: 3g of sodium lactate, 2g of sodium bicarbonate, 1g of sodium chloride, 0.5g of magnesium chloride hexahydrate, 0.015g of calcium chloride dihydrate, 0.5g of yeast extract, 0.3g of ammonium chloride, 0.3g of potassium chloride, 0.2g of potassium dihydrogen phosphate, 1mL of trace element liquid and 1000mL of distilled water.
Domestication degradation culture medium: ammonium chloride 0.5g, sodium thiosulfate 2.0g, potassium dihydrogen phosphate 5.44g, sodium pyruvate 0.5g, calcium chloride dihydrate 0.032g, magnesium chloride dihydrate 0.18g, cysteine 0.4g, trace element liquid 5mL, vitamin liquid 20mL (filtered and sterilized separately and added during use) and distilled water 1000mL.2,4, 6-trichlorophenol is gradually increased to 100mg/L from 10mg/L per 10mg/L level according to domestication requirement, and is added after sterilization.
Degradation screening plate medium: agar is added according to 1.5-2% based on acclimatization medium.
The liquid nutrient media used in this example included: 10g of peptone, 3g of beef extract, 5g of sodium chloride and 1000mL of distilled water, and 15-20g of agar are added into the liquid culture medium to obtain a nutrient agar culture medium plate.
The culture medium is prepared by mixing the above materials in conventional method, and sterilizing at 121deg.C for 15min.
The microelement liquid comprises the following components: 1g of sodium chloride, 0.1g of zinc chloride, 0.01g of boric acid, 0.12g of nickel chloride hexahydrate, 1.35g of ferric chloride hexahydrate, 0.1g of manganese chloride tetrahydrate, 12.8g of nitrilotriacetic acid, 0.024g of cobalt chloride hexahydrate, 0.25g of cupric chloride dihydrate, 0.1g of calcium chloride dihydrate, 0.024g of sodium molybdate dihydrate, 0.026g of sodium thiosulfate and 1000mL of distilled water.
The vitamin liquid comprises the following components: 5.0mg of para aminobenzoic acid, 5.0mg of vitamin B, 10.0mg of vitamin B, 9.0 mg of vitamin B, 0.1mg of vitamin B, 1L of distilled water and a 0.22 mu m sterilization filter.
Example 2 screening of 2,4, 6-trichlorophenol degrading Strain
The sludge sample adopted by the invention is from oilfield sludge of an extended oilfield somewhere in elm city of Shaanxi province.
(1) Sludge activation and gradual domestication-alternate culture
Mixing appropriate amount of sludge with 50 times of sterilized activation culture medium, placing in serum bottle, charging sterile nitrogen, covering with rubber plug, and standing at 37deg.C for 5 days. Inoculating activated sludge culture into domesticated degradation culture medium according to 20% ratio, wherein the initial concentration of 2,4, 6-trichlorophenol is 10mg/L, charging sterile nitrogen, sealing, and anaerobic culturing in anaerobic incubator at 37deg.C for 5 days. Inoculating the obtained culture solution into a liquid nutrient culture medium according to the proportion of 20%, and performing shaking-table aerobic culture at 37 ℃ and 120r/min for 24 hours. Inoculating the obtained culture solution into a domestication degradation culture medium according to the proportion of 20%, performing secondary domestication, wherein the concentration of 2,4, 6-trichlorophenol is 20mg/L, charging sterile nitrogen, sealing, performing anaerobic culture in an anaerobic incubator at 37 ℃ for 5 days, inoculating the obtained culture solution into a liquid nutrition culture medium according to the proportion of 20%, and performing shaking table aerobic culture at 37 ℃ for 24 hours at 120 r/min. And then continuously carrying out domestication of 2,4, 6-trichlorophenol with gradually increased concentration until the domestication concentration reaches 100mg/L, and carrying out aerobic culture of the liquid nutrient medium between each two stages of domestications.
(2) Separation and screening of Enterobacter cloacae OST-12
The sludge culture solution of gradual domestication-alternate culture is diluted and coated on a degradation flat plate containing 100mg/L2,4, 6-trichlorophenol degradation screening flat plate culture medium, anaerobic box culture is carried out at 37 ℃, after bacterial drop is grown, 8 single bacterial colonies are picked on the flat plate and placed on a nutrient agar culture medium flat plate for aerobic culture for 24 hours, bacterial colonies are respectively picked into a domestication degradation culture medium containing 100mg/L2,4, 6-trichlorophenol for primary culture, sterile nitrogen is filled, the anaerobic box culture is carried out at 37 ℃ for 5 days, supernatant is removed by centrifugation, the anaerobic culture is carried out in a domestication degradation culture medium containing 100mg/L2,4, 6-trichlorophenol which is fresh, anaerobic culture is continued for 5 days, supernatant is removed by centrifugation, anaerobic culture is carried out in a degradation culture medium containing 100mg/L2,4, 6-trichlorophenol which is fresh, and the degradation rate of 2,4, 6-trichlorophenol of the last stage is measured by liquid phase after continuous anaerobic constant temperature culture is carried out for three stages, as shown in figure 1. And selecting one strain with the best degradation effect from 8 strains, namely OST-12.
The method for measuring the 2,4, 6-trichlorophenol in the separation screening comprises the following steps: using a dry sterile syringe, 2mL of the culture broth was centrifuged at 12000r/min, and the supernatant was passed through a 0.45 μm aqueous membrane and analyzed by HPLC. Mobile phase: 2% acetic acid: methanol=3:7 (v/v), flow rate 1mL/min, sample injection amount 10. Mu.L, detection wavelength 290nm, column C-18, column temperature 30 ℃.
The degradation rate is calculated as follows:
2,4, 6-trichlorophenol degradation rate = 2,4, 6-trichlorophenol determination content/100 (mg/L) in culture solution after separation and screening continuous anaerobic constant temperature culture
Example 3 bacterial strain morphology and molecular characterization
In this example, the strain morphology and molecular sequence of Enterobacter cloacae OST-12 prepared in example 2 were identified, and the nutrient agar medium plate used in this example was the same medium as in example 1.
The strain OST-12 is coated on a nutrient agar culture medium plate, and cultured for 12 hours at 37 ℃ and has the morphology shown in figure 2, namely, mucus-type colonies, round, convex and neat in edge. Gram staining under microscope to gram negative Brevibacterium, no spores.
Extracting 16SrDNA of the strain OST-12 by using a 16S rDNA bacterial strain identification PCR kit, carrying out PCR amplification, and delivering an amplified product to Shanghai worker for sequencing to obtain the 16S rRNA nucleotide sequence of the strain OST-12, which is shown in a gene sequence table of SEQ ID No. 1. Then, a blast tool in NCBI website is utilized to search homologous sequences, clustalx is utilized to compare, mega is utilized to construct a phylogenetic tree, as shown in a figure 3, the result shows that the strain has the closest evolutionary distance with enterobacter cloacae, the sequence similarity reaches more than 99%, and the strain is named enterobacter cloacae OST-12.
Example 4 application detection test
In this example, the degradation efficiency of 2,4, 6-trichlorophenol by Enterobacter cloacae OST-12 prepared in example 1 was tested, and the liquid nutrient medium and the liquid acclimation degradation medium used in this example were the same as those used in example 1.
The 96-well microplates are used for carrying out various carbon source utilization characteristics conditions of the strain OST-12, the test result is shown in figure 4, the metabolic preference of the strain in the early stage is mainly concentrated on alcohols, acids and esters, and after 24 hours of culture, the utilization rate of amino acids, saccharides and amine carbon sources begins to rise, wherein the utilization rate of the amine carbon sources is continuously higher within 96 hours. The strain OST-12 is sensitive to carbohydrate and amino acid carbon sources through experiments, so that the strain OST-12 can be used as an electron donor in the later period and is beneficial to dechlorination.
Activating strain OST-12 at 37 ℃ with nutrient agar slant for 12h, inoculating into liquid nutrient medium, shaking at 37 ℃ with speed of 12r/min for 18h, centrifuging, pouring into sterilized liquid domestication degradation medium containing 2,4, 6-trichlorophenol 100mg/L, charging sterile nitrogen gas, sealing, standing for culturing in anaerobic culture at 37 ℃ for 24h, sucking 2mL of culture solution, centrifuging 12000r/min, taking supernatant, passing through water phase membrane of 0.45 μm, and testing degradation rate of 2,4, 6-trichlorophenol by liquid phase machine to form degradation curve of strain OST-12 to 2,4, 6-trichlorophenol, as shown in figure 5.
According to figure 5, the degradation rate of enterobacter cloacae OST-12 to 2,4, 6-trichlorophenol prepared by the invention is stabilized to the highest level in 7d, and the highest degradation rate can reach 90.68+/-5%.
The present invention may be better implemented as described above, and the above examples are merely illustrative of preferred embodiments of the present invention and not intended to limit the scope of the present invention, and various changes and modifications made by those skilled in the art to the technical solution of the present invention should fall within the scope of protection defined by the present invention without departing from the spirit of the design of the present invention.

Claims (2)

1. Enterobacter cloacaeEnterobacter cloacae) The method is characterized in that the preservation number of the enterobacter cloacae in China center for type culture collection is CCTCC NO: m20221208.
2. Use of enterobacter cloacae according to claim 1 for degrading 2,4, 6-trichlorophenol.
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Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998049106A1 (en) * 1997-04-25 1998-11-05 The University Of Iowa Research Foundation Fe(o)-based bioremediation of aquifers contaminated with mixed wastes
CN101389221A (en) * 2005-01-05 2009-03-18 罗伯特·霍拉迪 Silver/water, silver gels and silver-based compositions, method for fabricating and using the same
CN103627657A (en) * 2013-11-11 2014-03-12 华南理工大学 Composite microflora for degrading lignin wastewater and preparation method thereof
JP2014128264A (en) * 2012-11-29 2014-07-10 Eiken Chemical Co Ltd Medium for detecting enterohemorrhagic e. coli
CN104651291A (en) * 2015-02-10 2015-05-27 中国科学院天津工业生物技术研究所 Recombinant strain for producing phenol and application of strain
CN106119151A (en) * 2016-06-21 2016-11-16 盐城工学院 Bacterial strain and method for screening and separating thereof for degraded 2,4,6 trichlorophenol, 2,4,6,-Ts
CN106460013A (en) * 2014-05-13 2017-02-22 赢创德固赛有限公司 Method of producing organic compounds
CN106479916A (en) * 2016-09-29 2017-03-08 普洱学院 A kind of enterobacter cloacae bacterial strain and its application
CN113549565A (en) * 2021-03-25 2021-10-26 广州紫科环保科技股份有限公司 Enterobacter cloacae with benzene series degradation capability and application thereof
CN114853180A (en) * 2022-03-23 2022-08-05 陕西省微生物研究所 Biological agent, degradation agent, system and application for deeply degrading polychlorinated phenol

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4126809A2 (en) * 2020-04-01 2023-02-08 Cornell University Compositions and methods for inhibiting vibrio infection

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998049106A1 (en) * 1997-04-25 1998-11-05 The University Of Iowa Research Foundation Fe(o)-based bioremediation of aquifers contaminated with mixed wastes
CN101389221A (en) * 2005-01-05 2009-03-18 罗伯特·霍拉迪 Silver/water, silver gels and silver-based compositions, method for fabricating and using the same
JP2014128264A (en) * 2012-11-29 2014-07-10 Eiken Chemical Co Ltd Medium for detecting enterohemorrhagic e. coli
CN103627657A (en) * 2013-11-11 2014-03-12 华南理工大学 Composite microflora for degrading lignin wastewater and preparation method thereof
CN106460013A (en) * 2014-05-13 2017-02-22 赢创德固赛有限公司 Method of producing organic compounds
CN104651291A (en) * 2015-02-10 2015-05-27 中国科学院天津工业生物技术研究所 Recombinant strain for producing phenol and application of strain
CN106119151A (en) * 2016-06-21 2016-11-16 盐城工学院 Bacterial strain and method for screening and separating thereof for degraded 2,4,6 trichlorophenol, 2,4,6,-Ts
CN106479916A (en) * 2016-09-29 2017-03-08 普洱学院 A kind of enterobacter cloacae bacterial strain and its application
CN113549565A (en) * 2021-03-25 2021-10-26 广州紫科环保科技股份有限公司 Enterobacter cloacae with benzene series degradation capability and application thereof
CN114853180A (en) * 2022-03-23 2022-08-05 陕西省微生物研究所 Biological agent, degradation agent, system and application for deeply degrading polychlorinated phenol

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
Chun Chin Wang等.Biodegradation of 2,4,6-trichlorophenol in the presence of primary substrate by immobilized pure culture bacteria.Chemosphere.2000,第41卷(第12期),1873-1879. *
Reduced graphene oxide-supported metal organic framework as a synergistic catalyst for enhanced performance on persulfate induced degradation of trichlorophenol;Yanyan Xu等;Chemosphere;1-8 *
微氧条件下固定化颗粒污泥的氯酚降解及菌群结构;陈元彩;陈竹;蓝惠霞;付时雨;郭静;;华南理工大学学报(自然科学版)(09);128-132 *
氯酚菌的筛选及其对氯酚的降解;陈元彩, 蓝惠霞, 詹怀宇, 陈中豪;中国造纸(01);18-21 *
酚类有机污染物降解菌研究与应用;孙蕾;李卫忠;;甘肃科技纵横(01);31-32 *

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