CN110791450B - Strain Am101 capable of degrading multiple beta-lactam antibiotics and application thereof - Google Patents

Strain Am101 capable of degrading multiple beta-lactam antibiotics and application thereof Download PDF

Info

Publication number
CN110791450B
CN110791450B CN201911161751.0A CN201911161751A CN110791450B CN 110791450 B CN110791450 B CN 110791450B CN 201911161751 A CN201911161751 A CN 201911161751A CN 110791450 B CN110791450 B CN 110791450B
Authority
CN
China
Prior art keywords
strain
beta
lactam antibiotics
degradation
degrading
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201911161751.0A
Other languages
Chinese (zh)
Other versions
CN110791450A (en
Inventor
李艳菊
王梓诺
顾金刚
杨正礼
张爱萍
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Beijing Institute of Technology BIT
Original Assignee
Beijing Institute of Technology BIT
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Beijing Institute of Technology BIT filed Critical Beijing Institute of Technology BIT
Priority to CN201911161751.0A priority Critical patent/CN110791450B/en
Publication of CN110791450A publication Critical patent/CN110791450A/en
Application granted granted Critical
Publication of CN110791450B publication Critical patent/CN110791450B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • A—HUMAN NECESSITIES
    • A62—LIFE-SAVING; FIRE-FIGHTING
    • A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
    • A62D3/00—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances
    • A62D3/02—Processes for making harmful chemical substances harmless or less harmful, by effecting a chemical change in the substances by biological methods, i.e. processes using enzymes or microorganisms
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
    • 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
    • A—HUMAN NECESSITIES
    • A62—LIFE-SAVING; FIRE-FIGHTING
    • A62D—CHEMICAL MEANS FOR EXTINGUISHING FIRES OR FOR COMBATING OR PROTECTING AGAINST HARMFUL CHEMICAL AGENTS; CHEMICAL MATERIALS FOR USE IN BREATHING APPARATUS
    • A62D2101/00—Harmful chemical substances made harmless, or less harmful, by effecting chemical change
    • A62D2101/20—Organic substances
    • A62D2101/28—Organic substances containing oxygen, sulfur, selenium or tellurium, i.e. chalcogen
    • 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
    • C02F2101/38—Organic compounds containing nitrogen

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Environmental & Geological Engineering (AREA)
  • Hydrology & Water Resources (AREA)
  • Business, Economics & Management (AREA)
  • General Chemical & Material Sciences (AREA)
  • Water Supply & Treatment (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Toxicology (AREA)
  • Emergency Management (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Soil Sciences (AREA)
  • Mycology (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a Bacillus tequilensis Am101 capable of degrading various beta-lactam antibiotics and application thereof. The strain is stored in China general microbiological culture Collection center (CGMCC) at 11 months and 18 days in 2019, and the preservation number is CGMCC NO. 18965. The strain Am101 can degrade various beta-lactam antibiotics simultaneously, and can efficiently degrade penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium. The strain Am101 can be applied to degradation treatment of beta-lactam antibiotics remained in livestock and poultry manure, can also be applied to remediation of water bodies and soil polluted by the beta-lactam antibiotics, and has wide application prospect.

Description

Strain Am101 capable of degrading multiple beta-lactam antibiotics and application thereof
Technical Field
The invention relates to a strain Bacillus tequilensis (Am 101) capable of degrading various beta-lactam antibiotics, belonging to the technical field of microorganisms.
Background
The beta-lactam antibiotics refer to a large class of antibiotics containing beta-lactam rings in molecular structures, including penicillins taking 6-APA as a mother nucleus, cephalosporins taking 7-ACA as a mother nucleus and the like, have the characteristics of wide antibacterial spectrum, strong antibacterial action, low price and the like, are widely applied to the fields of animal husbandry, fishery, clinical medicine and the like, and are the most widely used antibiotics in the prior art.
During the fermentation production process of the beta-lactam antibiotics, a large amount of bacterial residues and waste water are generated, and if the beta-lactam antibiotics are treated improperly and discharged into the environment, residues in soil, water and food can be caused; in the using process of the antibiotics, only a small part of the antibiotics can be metabolized by human bodies and animals, and most of the antibiotics can enter the environment along with excrement in the form of drug prototypes or metabolites, so that drug-resistant bacteria and resistance genes are easily induced to be generated, and the ecological environment and the human health are threatened. Therefore, the removal of the antibiotic residues in the environment has become a problem to be solved in sustainable development of antibiotic industry and environmental management.
The method for treating the residual antibiotics in the environment by biodegradation is an environment-friendly, low-consumption and high-efficiency method, and becomes one of effective approaches for treating antibiotic pollution. However, few strains capable of rapidly and efficiently degrading beta-lactam antibiotics are reported at present, and particularly, strains capable of simultaneously degrading multiple beta-lactam antibiotics are not reported. Therefore, the patent provides a strain Am101 capable of degrading various beta-lactam antibiotics simultaneously.
Disclosure of Invention
The invention aims to provide a strain Am101 capable of degrading various beta-lactam antibiotics and application thereof.
The beta-lactam antibiotic degradation strain Am101 provided by the invention is Bacillus tequilensis (Bacillus tequilensis) and is named as Bacillus tequilensis Am101(Bacillus tequilensis Am 101). The strain is stored in China general microbiological culture Collection center (CGMCC) at 11 months and 18 days in 2019, and the preservation number is CGMCC NO. 18965. And (4) storage address: xilu No.1, Beijing, Chaoyang, Beijing, and institute for microbiology, China academy of sciences.
The strain Am101 is obtained by screening livestock and poultry manure, has the capability of degrading beta-lactam antibiotics, is wide in degradation spectrum and high in degradation efficiency, and can degrade beta-lactam antibiotics such as penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium when the pH value is 5-9; in addition, the co-metabolism carbon source or co-metabolism nitrogen source is added, so that the accelerated degradation of penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium antibiotics can be promoted, and the strain Am101 can be applied to the degradation treatment of beta-lactam antibiotics in the environment.
The strain Am101 can degrade penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium, can eliminate the biological toxicity of the four antibiotics and degradation products thereof to gram-positive indicator bacteria, has a detoxification effect, and can be used for degradation removal and harmless treatment of beta-lactam antibiotics in the environment.
The application of the strain Am101 or the fermentation liquor thereof in degradation of beta-lactam antibiotics is provided.
The strain Am101 or fermentation liquor thereof or a microbial inoculum containing the strain Am101 or the fermentation liquor is applied to the treatment of residual beta-lactam antibiotics in the degradation of livestock and poultry manure, bacterial residues and sludge solid waste, and the application to the remediation of water bodies or soil polluted by the beta-lactam antibiotics.
The invention has the beneficial effects that:
1. the strain Am101 has wide degradation spectrum, can simultaneously degrade various beta-lactam antibiotics such as penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium, and has high degradation speed and degradation rate of 100 percent.
2. The strain Am101 is obtained by screening livestock and poultry manure, and the strain and the prepared microbial inoculum thereof can be used for degradation treatment of residual beta-lactam antibiotics in the livestock and poultry manure, and are convenient to use.
3. The strain Am101 and the microbial inoculum thereof provided by the invention can be applied to remediation of water and soil polluted by beta-lactam antibiotics, and have the advantages of simple and convenient process, low cost and high efficiency.
Drawings
FIG. 1 is a colony morphology map of the strain Am 101.
FIG. 2 is a phylogenetic tree of strain Am 101.
FIG. 3 is a graph showing the growth of the strain Am101 on a beef extract peptone medium.
FIG. 4 is a degradation diagram of the strain Am101 with respect to penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium as the sole carbon and nitrogen source.
Detailed Description
Example 1:
screening of beta-lactam antibiotic degrading strain Am101
1. Material preparation
The bacteria sample source is as follows: fresh pig manure and chicken manure of certain livestock and poultry breeding enterprises.
Antibiotics: penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium.
Beef extract peptone medium: 5.0g of beef extract, 10.0g of peptone, 5.0g of NaCl, distilled water, pH7.4-7.6, and sterilizing at 121 ℃ for 15 min. Agar is added into the solid culture medium by 10g-15 g.
Culture medium II: 1.5g of beef extract, 6.0g of peptone, 6.0g of yeast extract powder, 1.0g of glucose and 1L of water, and sterilizing at 115 ℃ for 20 min. Agar is added into the solid culture medium by 10g-15 g.
Inorganic salt culture medium: (NH)4)2NO4 2.0g，K2HPO4 0.5g，KH2PO4 0.5g，MgSO4·7H2O0.5g，NaCl 0.2g，CaCl20.1g，MnSO4·H2O 0.01g，FeSO4·7H2O0.01 g and distilled water 1L, sterilizing at 121 deg.C for 15-20 min. Agar is added into the solid culture medium by 10g-15 g.
Martin's medium: KH (Perkin Elmer)2PO4 1.0g，MgSO4·7H20.5g of O, 10.0g of glucose, 5.0g of peptone and 1L of distilled water, wherein the pH is natural, and the sterilization is carried out for 15min at 115 ℃. Agar is added into the solid culture medium by 10g-15 g.
2. Strain screening
(1) Strain acclimation
Liquid domestication: taking pig manure or chicken manure according to the proportion of 5 percent (weight ratio), adding the pig manure or the chicken manure into an inorganic salt liquid culture medium which takes beta-lactam antibiotics as the only carbon-nitrogen source and has the contents of 50mg/L, 100mg/L and 150mg/L respectively, wherein penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium are added independently in an antibiotic adding mode, and the 4 antibiotics are added according to the ratio of 1:1:1:1, are added at 30 ℃, are transferred once every 48 hours by a 160-inch shaker at 180r/min, are transferred into an inorganic salt culture medium with the same concentration, and are transferred for 2-3 times for later use.
Solid domestication: by adopting a composting method, 200g of fresh pig manure is mixed with different beta-lactam antibiotics, the concentration of the antibiotics is set to be 50mg/Kg and 100mg/Kg, the addition mode of 4 beta-lactam antibiotics is the same as that of the liquid domestication, and the 4 antibiotics and the liquid domestication are independently added according to the ratio of 1:1:1: 1. Culturing at constant temperature of 30 ℃, and sampling after one week. Taking 5g of domesticated pig manure sample, adding an inorganic salt liquid culture medium with antibiotic as the only carbon-nitrogen source and the contents of 50mg/L and 100mg/L respectively, and carrying out shake cultivation at 30 ℃, 160-jar fermentation and 180r/min for 48h for later use.
(2) Separating and purifying strains
Respectively collecting the bacteria liquid enriched in the inorganic salt liquid culture medium in the step (1), and using sterile normal saline according to the ratio of 10-1,10-2,10-3Diluting in gradient concentration, spreading 200ul of diluent on inorganic salt solid culture medium with the same antibiotic and carbon and nitrogen source as separation culture medium, culturing at 28-30 deg.C for 3d-7d, collecting single colony with obvious growth, purifying by scribing method to obtain different strains, and storing at 4 deg.C.
(3) Strain screening
Screening by adopting an antibacterial ring method. Inoculating the strain obtained in the step (2) into a beef extract peptone (bacteria) or Martin's (fungi) liquid culture medium for activated culture, then respectively inoculating the activated bacterial liquid into an inorganic salt liquid culture medium taking penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium as unique carbon-nitrogen sources according to the inoculation amount of 3-10 percent and into an inorganic salt liquid culture medium compounding the 4 antibiotics according to the ratio of 1:1:1:1, setting no-inoculated bacteria as a reference, carrying out shake culture under the conditions of 28-37 ℃ and 150-170 r/min, taking a sample once a day, measuring the size of a zone of inhibition of micrococcus luteus, continuously measuring 6d, selecting a strain with high degradation speed and high degradation rate, and obtaining a strain with wide degradation spectrum and capability of simultaneously degrading penicillin potassium, Amoxicillin, cefuroxime sodium, ceftiofur sodium 4 kinds of beta-lactam antibiotics strain Am 101.
Secondly, identifying strain morphology and molecular biology
1. Observation of morphological characteristics of bacterial colonies
Am101 bacterial colony is round, the edge is smooth and wet, the bacterial colony grows for 1d on beef extract peptone culture medium, the size is 0.1-0.2mm, gram is positive, the thallus is rod-shaped, and the bacterial colony morphology of the bacterial strain Am101 is shown in figure 1.
2. Molecular biological characterization of the Strain CM1
2.1 extraction of the genome
The genomic DNA of the strain Am101 is extracted by using a bacterial genomic DNA rapid extraction kit produced by Biotechnology engineering, Inc. The genomic DNA of the strain Am101 was then sent to Biotechnology engineering (Shanghai) GmbH for PCR amplification and sequencing.
2.2 sequencing results and phylogenetic Tree construction
The 16S rDNA gene sequence of the strain Am101 is 1420bp in length after sequencing. The 16S rDNA sequence of the strain Am101 was found to have high homology with some strains of Bacillus tequilensis with a similarity of up to 99.93% compared to the ezBioCloud database. Gene sequences of strains with close similarity were selected and phylogenetic trees were constructed using MEGA 6.0 software, see FIG. 2. FIG. 2 shows that the strain Am101 and the Bacillus tequilensis KCTC 13622 strain are gathered in the same branch, and the strain Am101 is identified as Bacillus tequilensis (Bacillus tequilensis) and named as Bacillus tequilensis Am101(Bacillus tequilensis Am101) by combining the morphological characteristics of colonies. The strain is preserved in China general microbiological culture Collection center (CGMCC) at 11 months and 18 days in 2019, and the preservation number is CGMCC NO. 18965. And (4) storage address: xilu No.1, Beijing, Chaoyang, Beijing, and institute for microbiology, China academy of sciences.
At present, no document reports that Bacillus tequilensis (Bacillus tequilensis) has the function of degrading beta-lactam antibiotics at home and abroad. Therefore, the strain Am101 is a new functional strain capable of degrading beta-lactam antibiotics.
Example 2:
growth and degradation characteristics of strain Am101
Growth characteristics of strain Am101
The strain Am101 is inoculated in different solid culture mediums such as beef extract peptone culture medium, Martin culture medium, potato culture medium, inorganic salt culture medium and the like, and the strain Am101 can grow, can also grow in beef extract peptone culture medium containing penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium at different concentrations of 50-150mg/L, and is easy to culture.
A single-factor test is adopted to research the suitable culture condition of the strain Am101, the setting experiment comprises the temperature of 25-55 ℃, the pH value of 5-9, the concentration of beta-lactam antibiotics of 50mg/L-150mg/L and the culture time of 3-7 days, and the strain Am101 can grow under the conditions of 25-55 ℃, the pH value of 5-9 and the concentration of beta-lactam antibiotics of 50mg/L-150 mg/L.
Inoculating the strain Am101 into a beef extract peptone liquid culture medium, placing the beef extract peptone liquid culture medium under the condition of 28-30 ℃, carrying out shaking culture at 180r/min at 150-.
Secondly, degrading beta-lactam antibiotics by using strain Am101
1. Degradation of beta-lactam antibiotics as unique carbon and nitrogen source by strain Am101
(1) Activating and culturing the strain Am 101: inoculating the strain Am101 into a beef extract peptone liquid culture medium, and culturing for 1d at 30-37 ℃ for later use.
(2) Activating and culturing indicator bacteria:
micrococcus luteus ACCC No.41016 was selected as an indicator bacterium. And inoculating micrococcus luteus stored on the test tube slant to a No. II liquid culture medium, and culturing in a constant-temperature shaking table at 30 ℃ and 160r/min for 24h for later use.
(3) Preparation of inorganic salt culture medium with beta-lactam antibiotics as unique carbon-nitrogen source
And respectively adding 100mg/L of penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium into the sterilized and cooled inorganic salt liquid culture medium to obtain the antibiotic unique carbon-nitrogen source inorganic salt liquid culture medium.
(4) Inoculation and culture
Taking 40ml of the Am101 bacterial liquid activated in the step (1), 8000r/min, centrifuging for 5min, discarding the supernatant, taking the precipitated thallus, suspending the thallus by using 40ml of sterilized 0.9% sterile physiological saline, inoculating the thallus into the unique carbon-nitrogen source liquid culture medium of the antibiotic in the step (3) according to the inoculation amount of 5-15%, setting the non-inoculated bacteria as CK control, performing shake culture at the temperature of 28-37 ℃ and the temperature of 150r/min-180r/min, taking samples once every day, measuring the size of the inhibition zone of the activated Am101 bacterial liquid on micrococcus luteus, and continuously measuring for 5 d.
(5) And (3) determining the residual quantity of antibiotics: taking 100 μ l of activated indicator micrococcus luteus (ACCC No.41016) 200 μ l, uniformly coating on a beef extract peptone solid medium, placing 3-6 filter paper sheets with the diameter of 6mm on the medium, respectively dripping 4 μ l of the antibiotic degradation liquid prepared in the step (4) on the filter paper sheets, placing at 28-37 ℃ for culturing for 24-48h, measuring the diameter of the inhibition zone by using a vernier caliper, and calculating the degradation rate A%, wherein the calculation formula is as follows:
A％＝(Cckn-Cn)/Cckn*100％
wherein, CcknThe diameter of the zone of inhibition, C, in the control groupnThe diameter of the inhibition zone of the experimental group is shown, and n represents the time (d) for collecting the degradation liquid.
The results are shown in FIG. 4. According to the determination results, when 4 beta-lactam antibiotics of penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium are used as the unique carbon-nitrogen source, the degradation rate of the strain Am101 on the antibiotics can reach 100% within 1-4d, the degradation spectrum is wide, and the degradation efficiency is high. Can be used for the degradation treatment of beta-lactam antibiotics in the environment and the biological repair of polluted water and soil.
In addition, 4 antibiotics were mixed in a ratio of 1:1:1:1, when the total content is 100mg/L, the degradation rate can reach 100% through the degradation of the strain Am101 according to the method experiment. The bacterial strain Am101 can degrade 4 antibiotics of penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium simultaneously. The strain Am101 can be used for degradation treatment and biological repair of various beta-lactam antibiotics such as penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium in the environment in the presence of the same time.
2. Suitable conditions for degrading beta-lactam antibiotics by using strain Am101
And (3) setting a single-factor test to study the influence of different co-metabolic carbon sources, nitrogen sources and different pH values on the degradation of the beta-lactam antibiotics by the strain Am 101.
The co-metabolism carbon source is selected from corn starch, wheat bran, oat flour, soluble starch, sucrose, glucose and rice hull, and the co-metabolism nitrogen source is selected from NH4NO3、NH4Cl、NaNO3Beef extract, peptone and yeast extract powder, and the total content of beta-lactam antibiotics is 100 mg/L. Experiments show that the accelerated degradation of the beta-lactam antibiotics with the total content of 100mg/L, such as penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium, can be promoted by respectively adding the co-metabolism carbon source or the co-metabolism nitrogen source, and the degradation rate can reach 100%. Therefore, the strain Am101 or the fermentation liquor thereof can be treated in combination with a co-metabolism mode when the strain Am101 or the fermentation liquor thereof is used for degrading the beta-lactam antibiotics in the environment.
The strain Am101 has strong adaptability and good detoxification effect on beta-lactam antibiotics. Experiments with different pH values show that 4 antibiotics of penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium can be degraded by the strain Am101 within the pH value range of 5-9. When the addition ratio of the 4 antibiotics is 1:1:1:1, and the total content is 100mg/L, the strain Am101 can degrade the beta-lactam antibiotics by 100 percent through the degradation of the strain Am 101. Moreover, the antibacterial research result shows that the bacterial strain degrades degradation products of four beta-lactam antibiotics, the inhibition zone to the indicator bacteria becomes zero, the toxicity of the antibiotics is degraded and removed, and the effect of harmless treatment is achieved.
The strain Am101 can degrade penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium, can eliminate the biotoxicity of the four antibiotics and degradation products thereof to gram-positive indicator bacteria (micrococcus luteus), and has a biological detoxification effect, and the strain Am101 or fermentation liquor thereof or a microbial inoculum prepared from the strain Am101 or the fermentation liquor thereof can be used for biodegradation and harmless treatment of beta-lactam antibiotics such as penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium in the environments such as water, soil and the like.
3. Strain degradation beta-lactam antibiotics in livestock and poultry manure
Inoculating the strain Am101 into a beef extract peptone liquid culture medium for culture to obtain a fermentation broth, then respectively inoculating the fermentation broth into pig manure or chicken manure added with 4 beta-lactam antibiotics including penicillin potassium, amoxicillin, cefuroxime sodium and ceftiofur sodium according to the inoculation amount of 3-10%, carrying out solid fermentation for 7-10 days, and detecting the residual condition of the antibiotics by a bacteriostatic colony method. The detection result shows that the beta-lactam antibiotics in the chicken manure and the pig manure are well degraded. The inhibition zone of the chicken manure and the extracting solution thereof to the micrococcus luteus of the indicator bacterium becomes zero, which indicates that the biological toxicity of the above four beta-lactam antibiotics and the degradation products thereof to the micrococcus luteus is removed, thereby achieving the effect of harmless treatment. The strain Am101 or fermentation liquor thereof or a microbial inoculum prepared from the strain Am101 or the fermentation liquor thereof can be applied to the production of the beta-lactam antibiotics or the treatment of wastes of enterprises such as livestock and poultry manure, bacterial residues and sludge, and can also be applied to the remediation of soil and water bodies polluted by the beta-lactam antibiotics in the environment.
The above-mentioned examples are only preferred embodiments of the present invention, and are further additions and descriptions for the embodiment and the advantageous effects of the present invention, and it should be understood that the present invention is not limited to the specific embodiments illustrated, and any modification, replacement, etc. of the specific embodiments of the present invention by those skilled in the art are within the protection scope of the present invention.
Sequence listing
<110> Beijing university of science and technology
<120> strain Am101 capable of degrading various beta-lactam antibiotics and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1420
<212> DNA
<213> Bacillus tequilensis (Bacillus tequilensis)
<400> 1
gcaagtcgag cggacagatg ggagcttgct ccctgatgtt agcggcggac gggtgagtaa 60
cacgtgggta acctgcctgt aagactggga taactccggg aaaccggggc taataccgga 120
tggttgtttg aaccgcatgg ttcaaacata aaaggtggct tcggctacca cttacagatg 180
gacccgcggc gcattagcta gttggtgagg taacggctca ccaaggcaac gatgcgtagc 240
cgacctgaga gggtgatcgg ccacactggg actgagacac ggcccagact cctacgggag 300
gcagcagtag ggaatcttcc gcaatggacg aaagtctgac ggagcaacgc cgcgtgagtg 360
atgaaggttt tcggatcgta aagctctgtt gttagggaag aacaagtacc gttcgaatag 420
ggcggtacct tgacggtacc taaccagaaa gccacggcta actacgtgcc agcagccgcg 480
gtaatacgta ggtggcaagc gttgtccgga attattgggc gtaaagggct cgcaggcggt 540
ttcttaagtc tgatgtgaaa gcccccggct caaccgggga gggtcattgg aaactgggga 600
acttgagtgc agaagaggag agtggaattc cacgtgtagc ggtgaaatgc gtagagatgt 660
ggaggaacac cagtggcgaa ggcgactctc tggtctgtaa ctgacgctga ggagcgaaag 720
cgtggggagc gaacaggatt agataccctg gtagtccacg ccgtaaacga tgagtgctaa 780
gtgttagggg gtttccgccc cttagtgctg cagctaacgc attaagcact ccgcctgggg 840
agtacggtcg caagactgaa actcaaagga attgacgggg gcccgcacaa gcggtggagc 900
atgtggttta attcgaagca acgcgaagaa ccttaccagg tcttgacatc ctctgacaat 960
cctagagata ggacgtcccc ttcgggggca gagtgacagg tggtgcatgg ttgtcgtcag 1020
ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caacccttga tcttagttgc 1080
cagcattcag ttgggcactc taaggtgact gccggtgaca aaccggagga aggtggggat 1140
gacgtcaaat catcatgccc cttatgacct gggctacaca cgtgctacaa tggacagaac 1200
aaagggcagc gaaaccgcga ggttaagcca atcccacaaa tctgttctca gttcggatcg 1260
cagtctgcaa ctcgactgcg tgaagctgga atcgctagta atcgcggatc agcatgccgc 1320
ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac accacgagag tttgtaacac 1380
ccgaagtcgg tgaggtaacc ttttaggagc cagccgccga 1420

Claims (7)

1. Bacillus tequilensis for degrading beta-lactam antibioticsBacillus tequilensis) The strain Am101 has the preservation number of CGMCC NO.18965, and is preserved in China general microbiological culture Collection center in 11-18.2019.
2. The use of the Bacillus tequilensis strain Am101 of claim 1 in the degradation of β -lactam antibiotics.
3. The use according to claim 2, wherein the Bacillus tequilensis strain Am101 uses a co-metabolized carbon source or a co-metabolized nitrogen source to accelerate the degradation of the β -lactam antibiotic.
4. The use of claim 2, wherein the Bacillus tequilensis strain Am101 is used for degrading and eliminating the biological toxicity of the beta-lactam antibiotics and degradation products thereof to gram-positive indicator bacteria, thereby achieving the effects of detoxification and harmless treatment.
5. The use of the strain Am101 or the fermentation liquid thereof, or the microbial inoculum containing the strain Am101 or the fermentation liquid thereof according to claim 2, for degrading the residual beta-lactam antibiotics.
6. Use according to claim 5, characterized in that the antibiotic residual waste is selected from the group consisting of poultry manure, sludge and antibiotic residues.
7. The use of the strain Am101 or the fermentation liquid thereof or the microbial inoculum containing the strain Am101 or the fermentation liquid thereof as the raw material of the preparation of the fertilizer according to claim 2, for degrading and restoring the soil or water body polluted by the beta-lactam antibiotics.
CN201911161751.0A 2019-11-22 2019-11-22 Strain Am101 capable of degrading multiple beta-lactam antibiotics and application thereof Active CN110791450B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201911161751.0A CN110791450B (en) 2019-11-22 2019-11-22 Strain Am101 capable of degrading multiple beta-lactam antibiotics and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201911161751.0A CN110791450B (en) 2019-11-22 2019-11-22 Strain Am101 capable of degrading multiple beta-lactam antibiotics and application thereof

Publications (2)

Publication Number Publication Date
CN110791450A CN110791450A (en) 2020-02-14
CN110791450B true CN110791450B (en) 2021-01-19

Family

ID=69445894

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201911161751.0A Active CN110791450B (en) 2019-11-22 2019-11-22 Strain Am101 capable of degrading multiple beta-lactam antibiotics and application thereof

Country Status (1)

Country Link
CN (1) CN110791450B (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114058529B (en) * 2020-07-30 2023-05-26 中国石油天然气股份有限公司 Oily pollutant degradation strain ZT2, microbial agent and application
CN115181691B (en) * 2022-06-21 2024-04-19 中国科学院烟台海岸带研究所 Bacillus DB-Q3 and application thereof
CN115141770B (en) * 2022-06-27 2023-08-29 安阳工学院 Bacillus tequilensis H1 capable of degrading COD in livestock wastewater and application thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2836589B1 (en) * 2012-04-13 2018-08-15 Chr. Hansen A/S Antibiotic sensitive bacillus strains having antimicrobial effect against e. coli and clostridium perfringens and having high sporulation capacity
CN107858312B (en) * 2017-12-07 2020-11-17 北京理工大学 Beta-lactam antibiotic degradation strain PGB1 and application thereof

Also Published As

Publication number Publication date
CN110791450A (en) 2020-02-14

Similar Documents

Publication Publication Date Title
CN110791450B (en) Strain Am101 capable of degrading multiple beta-lactam antibiotics and application thereof
WO2015184935A1 (en) Efficient bottom treatment bacillus, composite bottom treatment inoculant prepared using same and applications thereof
CN109576187B (en) Cyanide degradation strain and method for degrading cyanide by using same
CN110055187B (en) Pseudomonas proteorum and application thereof
CN111733113B (en) COD (chemical oxygen demand) degrading strain and application thereof
CN109837230B (en) Bacillus amyloliquefaciens Y1711, culture method and application thereof
CN112011486B (en) Synchronous nitrification and denitrification bacillus subtilis K8 and application thereof
CN108865940B (en) Heterotrophic nitrification-aerobic denitrification bacillus and composite bacterial preparation thereof
CN110656066B (en) Acinetobacter strain for shortcut nitrification and denitrification and application thereof
Shen et al. Combination of heterotrophic nitrifying bacterium and duckweed (Lemna gibba L.) enhances ammonium nitrogen removal efficiency in aquaculture water via mutual growth promotion
CN113862174B (en) Antibiotic degrading bacterium and application thereof in soil pollution remediation
CN114621885B (en) Bacillus subtilis for efficiently removing ammoniacal nitrogen and nitrite nitrogen and application of bacillus subtilis in aquaculture
CN115449489A (en) Oil reducing bacteria and composite microbial inoculum thereof, preparation method and application
CN107858312B (en) Beta-lactam antibiotic degradation strain PGB1 and application thereof
CN117448206A (en) Bacillus pumilus and application thereof in culture water body regulation
CN115386520B (en) Rhodococcus pyridine-philic RL-GZ01 strain and application thereof
CN111454861A (en) Bacillus amyloliquefaciens for efficiently purifying sewage, microbial agent and application
CN117467555A (en) Bacillus pricklyash strain and application thereof
CN106591181B (en) A kind of Mysore arthrobacterium and its application in purifying sea water cultivation nitrogenous effluent
CN114231448B (en) Burkholderia contaminated with indole degradation capability and application thereof
CN113755400B (en) Bacillus cereus for degrading aureomycin and application thereof
CN113846038A (en) Arthrobacter strain and application thereof in degradation of aristolochic acid
CN112479391A (en) Preparation method of degradation product of well site environment-friendly toilet and method for treating water-based solid waste
CN108441437B (en) Complex microbial inoculant and application thereof
JP5046665B2 (en) Novel microorganism and drainage purification method using the same

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant