CN108635371B - Application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm - Google Patents
Application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm Download PDFInfo
- Publication number
- CN108635371B CN108635371B CN201810054101.5A CN201810054101A CN108635371B CN 108635371 B CN108635371 B CN 108635371B CN 201810054101 A CN201810054101 A CN 201810054101A CN 108635371 B CN108635371 B CN 108635371B
- Authority
- CN
- China
- Prior art keywords
- quantum dots
- carbon
- fluorescent quantum
- escherichia coli
- formation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 229910052799 carbon Inorganic materials 0.000 title claims abstract description 43
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 41
- 239000002096 quantum dot Substances 0.000 title claims abstract description 38
- 241000588724 Escherichia coli Species 0.000 title abstract description 23
- 230000015572 biosynthetic process Effects 0.000 title abstract description 12
- 230000005764 inhibitory process Effects 0.000 title abstract description 7
- 239000012528 membrane Substances 0.000 claims description 11
- 241001052560 Thallis Species 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 238000001027 hydrothermal synthesis Methods 0.000 claims description 10
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 9
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 9
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 241000186660 Lactobacillus Species 0.000 claims description 7
- 229940039696 lactobacillus Drugs 0.000 claims description 7
- 239000002245 particle Substances 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 4
- 238000009630 liquid culture Methods 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 230000035484 reaction time Effects 0.000 claims description 3
- 230000003068 static effect Effects 0.000 claims description 3
- 239000000725 suspension Substances 0.000 claims description 2
- 239000006872 mrs medium Substances 0.000 claims 1
- -1 polytetrafluoroethylene Polymers 0.000 claims 1
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims 1
- 239000004810 polytetrafluoroethylene Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 7
- 235000013305 food Nutrition 0.000 abstract description 6
- 230000003214 anti-biofilm Effects 0.000 abstract description 3
- 231100000956 nontoxicity Toxicity 0.000 abstract description 3
- 238000012545 processing Methods 0.000 abstract description 3
- 230000002147 killing effect Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 15
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 11
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 11
- 238000002360 preparation method Methods 0.000 description 10
- 230000005284 excitation Effects 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 4
- 230000005540 biological transmission Effects 0.000 description 4
- 235000011187 glycerol Nutrition 0.000 description 4
- 239000002086 nanomaterial Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000032770 biofilm formation Effects 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- 238000002329 infrared spectrum Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 238000004020 luminiscence type Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000002715 modification method Methods 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 208000017667 Chronic Disease Diseases 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 239000012901 Milli-Q water Substances 0.000 description 1
- 235000005811 Viola adunca Nutrition 0.000 description 1
- 240000009038 Viola odorata Species 0.000 description 1
- 235000013487 Viola odorata Nutrition 0.000 description 1
- 235000002254 Viola papilionacea Nutrition 0.000 description 1
- 244000172533 Viola sororia Species 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 238000012864 cross contamination Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000011866 long-term treatment Methods 0.000 description 1
- 210000005265 lung cell Anatomy 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/44—Elemental carbon, e.g. charcoal, carbon black
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Inorganic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Pest Control & Pesticides (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Microbiology (AREA)
- Agronomy & Crop Science (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides an application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm, wherein the carbon fluorescent quantum dots have excellent anti-biofilm activity and no toxicity to normal cells; provides an effective way for effectively killing the Escherichia coli biomembrane in food, medical supplies and food and medical processing environments.
Description
Technical Field
The invention belongs to the technical field of nano materials and biology, and particularly relates to application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm.
Background
Biofilm refers to a compact structure formed by microbial populations adsorbed to the surface of some objects or interacting with each other and embedded in an extracellular matrix composed of polysaccharides, proteins and nucleic acids (Nature Reviews Microbiology,14(9), 563-575). Bacterial biofilms are widely present in natural environments, medical devices and industrial equipment. The biological membrane has strong stress resistance and great harm (Current opinion in microbiology,16(5), 580-589). The formation of biofilms can cause cross-contamination of food, medical supplies, and food, medical processing environments and equipment (Biomaterials,2011,32(23): 5459-. The formation of biofilms increases the probability of disease and increases the difficulty of disease treatment (Science,301(5629), 105-. For example, biofilms increase the risk of chronic disease in humans, increase the difficulty of organ transplantation and even cause death in industry, and biofilms can damage various industrial facilities, such as filter clogging, pipeline corrosion, etc. (Nature nanotechnology,7(8), 530-. However, eliminating and inhibiting biofilm formation is currently a major challenge in the industry as well as the biomedical industry.
In the prior art, the effect of eliminating the biofilm by using antibiotics alone is often poor, and the intervention of surgical treatment is often needed, thereby causing high cost of long-term treatment (Cold Spring Harbor peroxides in media, 3(4), a 010306). And the development of the novel antibiotics is difficult and slow, and is far behind the emergence speed of drug-resistant microorganisms. Therefore, there is an urgent need to develop alternative strategies to combat biofilms. With the rapid development of nanobiotechnology, a large number of nanomaterials having unique biological characteristics have been developed and used to inhibit the formation of various biofilms. For example, nanoparticles containing metal/metal oxide nanoparticles (ACS nano,11 (9)), 9330-; although they have good anti-biofilm activity, nanomaterials are generally toxic to microorganisms themselves as well as human cells and have poor biocompatibility (ACS nano,2017,11(9): 9330-9339). In order to solve the problem, people adopt a chemical or physical modification method to improve the surface characteristics of the nano material so as to achieve the aim of effective biofilm resistance and no toxicity to cells; however, these modification methods are complicated in steps and are often not suitable for industrial production.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm; the carbon fluorescent quantum dots prepared by taking lactobacillus plantarum thalli as a raw material achieve the purpose of inhibiting an escherichia coli biological membrane; the carbon fluorescent quantum not only has excellent anti-biofilm activity, but also has no toxicity to normal cells.
In order to achieve the purpose, the technical scheme of the invention is as follows:
an application of carbon fluorescent quantum dots in inhibiting the formation of escherichia coli biofilm.
Preferably, the carbon fluorescent quantum dots are prepared by taking lactobacillus as a raw material.
The carbon fluorescent quantum dot is prepared by the following steps:
(1) inoculating lactobacillus into liquid culture medium, and standing for culture to obtain lactobacillus thallus.
(2) And (2) adding the thalli obtained in the step (1) into 20-50 mL of water for suspension, placing the thalli into a hydrothermal reaction kettle, placing the thalli into an oven for reaction, centrifuging to remove large particles, and filtering by using a 0.22-0.45-micrometer filter membrane to obtain the carbon fluorescent quantum dots.
Wherein the liquid culture medium in the step (1) is an MRS culture medium.
Further, in the step (1), the lactobacillus is lactobacillus plantarum (lactobacillus plantarum) preserved in a glycerin pipe, is separated from Yunnan pickle, has a strain number of LCC-605, is preserved in China center for type culture Collection, has a preservation number of CCTCC No. M2016491, and has an accession number of NCBI of: KX443590 with a preservation date of 2016, 9/18.
Preferably, the lactobacillus plantarum LCC-605 culture method is referred to the preparation method of the inventor's prior patent 201710025113.0, or is obtained by conventional means.
Further, in the step (1), the ratio of the lactobacillus inoculated in the liquid culture medium is 1-5%; the temperature of static culture is 25-37 ℃, and the time is 12-24 h; the centrifugal speed is 5000-8000 rpm, and the time is 10-15 min.
Preferably, the inoculation ratio of lactobacillus is 3%; (ii) a The temperature of static culture is 31 ℃, and the time is 12 h; the centrifugation speed was 5000rpm and the time was 10 min.
Preferably, in the step (2), the cell obtained in the step (1) is added to a water volume of 30mL and a filter pore size of 0.22. mu.m.
Further, in the step (2), the reaction temperature is 120-240 ℃, and the reaction time is 12-48 h; the centrifugal speed is 8000-12000 rpm, and the time is 10-15 min.
Preferably, in the step (2), the reaction temperature is 120 ℃, and the reaction time is 12 h; the centrifugation speed was 12000rpm for 10 min.
Preferably, in the step (2), the hydrothermal reaction kettle is a polytetrafluoroethylene-lined high-temperature high-pressure hydrothermal reaction kettle, and the volume of the hydrothermal reaction kettle is 100-150 mL.
Has the advantages that: compared with the prior art, the invention has the following remarkable advantages: the formation of the biological membrane is obviously reduced along with the increase of the concentration of the carbon fluorescent quantum dots, and when the concentration of the carbon fluorescent quantum dots reaches 3-6 mg/ml, the biological membrane for escherichia coli is completely killed, and meanwhile, the biological membrane is free of toxicity to normal cells; provides an effective way for effectively killing the Escherichia coli biomembrane in food, medical supplies and food and medical processing environments.
Drawings
FIG. 1 is a transmission electron microscope image of carbon fluorescent quantum dots;
FIG. 2 is a schematic diagram of the particle size distribution Frequency (Frequency) of carbon fluorescent quantum dots;
FIG. 3 is a schematic infrared spectrum of carbon fluorescent quantum dots;
FIG. 4 is a graph showing the results of zeta potential (zeta potential) of carbon fluorescent quantum dots;
FIG. 5 is a bar graph of the effect of carbon fluorescent quantum dot concentration on the remaining percentage of stained E.coli biofilm;
FIG. 6 is a 3D image of carbon fluorescence quantum dot treated Escherichia coli biofilm cultured for 72h laser confocal observation, with an excitation wavelength of 488nm and carbon dot concentrations of 0, 0.1, 0.8 and 3 mg/mL;
FIG. 7 is a bar graph of cell counts of carbon fluorescence quantum dots versus cell viability of free E.coli cells;
FIG. 8 is a graph showing the evaluation of the toxicity (cell viability) of carbon fluorescence quantum dots to animal cells.
Detailed Description
The technical solution of the present invention will be further described with reference to the following examples. The starting materials and reagents used in the present invention and the like may be obtained by ordinary commercial purchase or preparation by the prior art, unless otherwise specified.
Example 1 preparation of carbon fluorescent quantum dots (CDs-LP):
inoculating lactobacillus plantarum LCC-605 preserved in a glycerol tube into a liquid MRS culture medium according to the proportion of 5%, standing and culturing at 37 ℃ for 24h, and centrifuging the obtained fermentation liquor at 8000rpm for 15min to obtain thalli for later use. The cells were suspended in 50mL of water and placed in a 150mL hydrothermal reaction vessel. Placing the mixture in an oven at 240 ℃ to react for 48 h. And after the reaction is finished, removing residues, centrifuging at 10000rpm for 12min to remove large particles, and filtering the residual liquid by using a 0.35-micrometer filter membrane to obtain the carbon fluorescent quantum dots with the performances of multicolor luminescence and the like.
Example 2 preparation of carbon fluorescent quantum dots (CDs-LP):
inoculating lactobacillus plantarum LCC-605 preserved in a glycerol tube into a liquid MRS culture medium according to the proportion of 1%, standing and culturing at 25 ℃ for 12h, and centrifuging the obtained fermentation liquor at 5000rpm for 15min to obtain thalli for later use. The cells were suspended in 20mL of water and placed in a 100mL hydrothermal reaction vessel. Placing the mixture in an oven at 120 ℃ to react for 18 h. And after the reaction is finished, removing residues, centrifuging at 8000rpm for 15min to remove large particles, and filtering the residual liquid with a 0.45-micrometer filter membrane to obtain the carbon fluorescent quantum dots with multicolor luminescence and other performances.
Example 3 preparation of carbon fluorescent quantum dots (CDs-LP):
inoculating lactobacillus plantarum LCC-605 preserved in a glycerol tube into a liquid MRS culture medium according to the proportion of 3%, standing and culturing at 31 ℃ for 12h, and centrifuging the obtained fermentation liquor at 5000rpm for 10min to obtain thalli for later use. The cells were suspended in 30m L water and placed in a 100mL hydrothermal reaction vessel. Placing the mixture in an oven at 120 ℃ for reaction for 12 h. After the reaction, the residue was removed, and after centrifugation at 12000rpm for 10min to remove large particles, the remaining liquid was filtered through a 0.22 μm filter for use.
The transmission electron microscope observation of the carbon fluorescent quantum dots obtained by the preparation method comprises the following steps:
diluting the filtered carbon quantum dots by 10 times by using deionized water, dripping 10 mu L of the diluted carbon quantum dots on a 400-mesh copper net, and observing the carbon quantum dots by using a transmission electron microscope (JEM-2100, JEOL Ltd., Japan); the image results are shown in FIG. 1, and the quantitative results are shown in FIG. 2.
Infrared spectrum scanning of the carbon fluorescent quantum dots obtained by the preparation method comprises the following steps:
the lyophilized carbon quantum dots were measured for Absorbance (Absorbance) at different wavelengths (Wavenumber) using FTIR (Nicolet iS50, Thermo Scientific, USA) and the results are shown in FIG. 3.
Potential detection of the carbon fluorescent quantum dots obtained by the preparation method comprises the following steps:
the obtained fluorescent carbon quantum dots are placed in a Zeta potential (Zeta potential) detection cell, the electric potential of the fluorescent carbon quantum dots is detected by using a DLS instrument, and the detection result is shown in figure 4.
Fluorescence spectrum scanning of the carbon fluorescent quantum dots obtained by the preparation method comprises the following steps:
the excitation Wavelength is selected from the range of 300-520 nm, and the emission Wavelength (wavelet) and the fluorescence intensity (PL intensity) are measured every 40 nm.
The observation result of a transmission electron microscope in the attached figure 1 shows that the carbon quantum dots are approximately spherical in structure and are uniformly distributed; figure 2 the results show that the average particle Diameter (Diameter) is about 3 nm; the infrared spectrum scan of figure 3 shows: the carbon quantum dot contains functional groups such as N-H, C-H, C ═ O, C-O-C and C-N; the potential measurements of FIG. 4 show that the carbon quantum dots are negatively charged (-22 mV); the fluorescence spectrum scanning result shows that the carbon quantum dots emit blue-violet fluorescence under the irradiation of ultraviolet light, the fluorescence emission spectrum has excitation wavelength dependence, and the peak value of the emission wavelength shifts from 420 to 540nm along with the increase of the excitation wavelength. In summary, the carbon fluorescent quantum dots obtained in example 3 have properties such as multicolor luminescence.
Example 4 inhibition of E.coli biofilm formation by carbon fluorescent Quantum dots (CDs-LP)
Preparation of experimental group Escherichia coli biofilm
Inoculating an Escherichia coli strain (E.coli DH5 alpha (pLGZ-901) which is originated from China Center for Industrial Culture Collection (CICC) at 37 ℃ and 180rpm and cultured overnight) into 1/5 LB culture medium in a ratio of 1:100, adding CDs-LP with different concentrations (0, 0.01, 0.025, 0.05, 0.1, 0.2, 0.4, 0.8, 1,3 and 6mg/ml) as a control group without adding CDs-LP, transferring the control group to a 96-well plate (100 mu L/well), and statically culturing the inoculated well plate 96 at 28 ℃ for 1-5 days to obtain the Escherichia coli biomembrane of the experimental group with different concentrations of carbon dots.
The formation of the E.coli biofilm of the experimental groups was characterized:
confocal observation: washing the cultured biological membrane with PBS for three times, adding dead/live dye for dyeing, observing under a laser confocal microscope, wherein the excitation wavelength is 488nm, the emission wavelengths are 500nm and 635nm respectively, the confocal observation result is shown in figure 6, and the quantity of the biological membrane is reduced sharply along with the increase of the concentration;
crystal violet dyeing: heating cultured biomembrane at 60 deg.C for 1 hr, and staining with crystal violet10min, then washed with Milli-Q water until no color eluted. After removing the residual liquid, the stained biofilm was photographed and then eluted with an eluent (a mixed solution of 10% methanol and 7.5% acetic acid, solvent is water). Then reading the light absorption value at 595nm by using an enzyme reader, and increasing the concentration of the used carbon spots, namely OD595The gradual decrease shows that the formation of the biofilm is gradually reduced, and when the concentration of the CDs-LP reaches 3-6 mg/ml, the Escherichia coli biofilm is completely killed. The results are shown in FIG. 5 and Table 1.
TABLE 1 inhibition of E.coli biofilm formation by carbon fluorescent Quantum dots (CDs-LP)
Example 5 effect of carbon fluorescent quantum dots (CDs-LP) on growth of e.coli:
coli grown overnight was inoculated in 1/5 LB medium with (i.e., CDs-LP) or without (i.e., control) CDs-LP at a ratio of 1:100, and then transferred to 96-well plates (100. mu.L/well), and incubated in an incubator at 28 ℃ for 24h for plating cell counting, as shown in FIG. 7, the results of the experiment showed no significant difference in cell counting between the experimental group and the blank group, i.e., CDs-LP was not toxic to free E.coli.
Example 6 evaluation of safety of carbon fluorescent Quantum dots (CDs-LP)
Normal lung cells AT II were seeded in DMEM medium containing 10% FBS in 5% CO2The culture was carried out in a 37 ℃ incubator. Cultured cells were plated in 96-well plates (5X 10)4Individual cells/well) were cultured overnight and then treated with different concentrations of CDs-LP (0, 0.01, 0.05, 0.1, 0.5, 1,3 and 6mg/mL) for 24 h. Then adding 10 mu L of MTT with the concentration of 5mg/mL for culturing for 4h, then pouring out the liquid and adding 150 mu L of DMSO; the absorbance at 492nm was measured, and the results in FIG. 8 and Table 3 indicate that the CDs-LP is not toxic to cells at concentrations below 6 mg/mL.
TABLE 3 evaluation of cytotoxicity of carbon fluorescent Quantum dots (CDs-LP)
| CDs-LP(mg/ml) | 0 | 0.01 | 0.05 | 0.1 | 0.5 | 1 | 3 | 6 |
| Absorbance (OD)492) | 0.8116 | 0.8566 | 0.8358 | 0.7454 | 0.8577 | 0.8067 | 0.8016 | 0.8566 |
| Cell Activity (%) | 100 | 105.542 | 102.973 | 91.8452 | 105.673 | 99.3965 | 98.7669 | 105.54 |
Claims (4)
1. The carbon fluorescent quantum dot is prepared from lactobacillus plantarum LCC-605CCTCC No: M2016491 serving as a raw material, and is prepared by the following steps:
(1) inoculating lactobacillus plantarum LCC-605CCTCC No: M2016491 in a liquid culture medium according to a proportion of 1-5%, statically culturing, and centrifuging to obtain a thallus of lactobacillus;
(2) adding the thalli obtained in the step (1) into 20-50 mL of water for suspension, putting the thalli into a hydrothermal reaction kettle, placing the thalli into an oven for reaction, centrifuging to remove large particles, and filtering with a 0.22-0.45 mu m filter membrane to obtain the carbon fluorescent quantum dots;
in the step (1), the temperature of the static culture is 25-37 ℃, and the time is 12-24 h; the centrifugal speed is 5000-8000 rpm, and the time is 10-15 min.
2. Use according to claim 1, characterized in that: in the step (1), the liquid medium is an MRS medium.
3. Use according to claim 1, characterized in that: in the step (2), the reaction temperature is 120-240 ℃, and the reaction time is 12-48 h; the centrifugal speed is 8000-12000 rpm, and the time is 10-15 min.
4. Use according to claim 1, characterized in that: the hydrothermal reaction kettle is a high-temperature high-pressure hydrothermal reaction kettle with a polytetrafluoroethylene lining, and the volume of the reaction kettle is 100-150 mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810054101.5A CN108635371B (en) | 2018-01-19 | 2018-01-19 | Application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810054101.5A CN108635371B (en) | 2018-01-19 | 2018-01-19 | Application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108635371A CN108635371A (en) | 2018-10-12 |
| CN108635371B true CN108635371B (en) | 2020-12-25 |
Family
ID=63744067
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810054101.5A Active CN108635371B (en) | 2018-01-19 | 2018-01-19 | Application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108635371B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113413397A (en) * | 2021-06-10 | 2021-09-21 | 哈尔滨工业大学 | Application based on carbon quantum dots |
| CN114477138B (en) * | 2021-12-15 | 2023-07-07 | 浙江工业大学 | Preparation method of potato carbon quantum dot and degradable preservative film with high antibacterial activity |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058166B (en) * | 2017-01-13 | 2020-07-31 | 东南大学 | Lactobacillus plantarum for producing exopolysaccharides |
| CN107418566B (en) * | 2017-06-06 | 2020-06-30 | 东南大学 | Preparation method of carbon quantum dots and application of carbon quantum dots in biomembrane imaging |
-
2018
- 2018-01-19 CN CN201810054101.5A patent/CN108635371B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN108635371A (en) | 2018-10-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107418566B (en) | Preparation method of carbon quantum dots and application of carbon quantum dots in biomembrane imaging | |
| CN109207440B (en) | Vibrio bacteriophage and preparation method and application of bactericidal composition thereof | |
| Yunos et al. | Harvesting of microalgae (Chlorella sp.) from aquaculture bioflocs using an environmental-friendly chitosan-based bio-coagulant | |
| Chen et al. | Effects of soil pH, temperature and water content on the growth of Burkholderia pseudomallei | |
| CN107686832B (en) | Novel vibrio parahaemolyticus bacteriophage, and composition, preparation method and application thereof | |
| Chen et al. | Graphene oxide as an anaerobic membrane scaffold for the enhancement of B. adolescentis proliferation and antagonistic effects against pathogens E. coli and S. aureus | |
| CN108635371B (en) | Application of carbon fluorescent quantum dots in inhibition of formation of escherichia coli biofilm | |
| CN104830806A (en) | Salmonella bacteriophage being wide in lysis spectrum and bacterial inhibition application thereof | |
| CN108699533B (en) | Vibrio parahaemolyticus bacteriophage Vib-PAP-1 and use thereof for inhibiting proliferation of Vibrio parahaemolyticus | |
| CN108699532B (en) | Vibrio parahaemolyticus bacteriophage Vib-PAP-2 and use thereof for inhibiting proliferation of Vibrio parahaemolyticus | |
| KR101825439B1 (en) | Method for preparing Gram positive bacteria ghosts by the treatment with hydrochloric acid | |
| Shangguan et al. | Anti‐biofilm potential of kefir‐derived Lactobacillus paracasei L10 against Vibrio parahaemolyticus | |
| Kıvanç et al. | Biofilm formation of Candida Spp. isolated from the vagina and antibiofilm activities of lactic acid bacteria on the these Candida Isolates | |
| Abdolnabi et al. | Pathogenicity of Aeromonas hydrophila in giant freshwater prawn Macrobrachium rosenbergii, cultured in East Malaysia | |
| CN106754559A (en) | A kind of probiotics preparation method for effectively purifying water | |
| Sunitha et al. | Efficacy of probiotics in water quality and bacterial biochemical characterization of fish ponds | |
| KR101654253B1 (en) | Manufacturing method and the usage of eco-friendly aquatic fungicidal agent composed of the water containing detoxified sulfur for use in fish. | |
| WO2021023564A1 (en) | Marinomonas ef1 and rhodococcus ef1 for the production of secondary metabolites | |
| CN109022303B (en) | Composition with functions of dissolving algae and destroying microalgae air bags and application thereof | |
| Dayma et al. | Influence of low salinity stress on virulence and biofilm formation potential in Vibrio alginolyticus, isolated from the Gulf of Khambhat, Gujarat India | |
| CN111514308B (en) | PH-induced charge-inversion antibacterial gold nanorod and preparation method and application thereof | |
| Waturangi et al. | Antibiofilm activity of bacteria isolated from marine environment in Indonesia against Vibrio cholerae | |
| Mahjoub et al. | Inhibitory activity of native probiotic Bacillus vallismortis IS03 against pathogenic Vibrio harveyi under in vitro and in vivo conditions in Litopenaeus vannamei | |
| Stoyanova et al. | Bacterial Bulb Decay of Summer Snowflake/Leucojum Aestivum L. | |
| KR102612095B1 (en) | Anti-microbial composition comprising bioconversion product of Nostoc commune extract or extract thereof |
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 |