CN107557317B - Streptococcus mitis, application thereof and product using streptococcus mitis - Google Patents
Streptococcus mitis, application thereof and product using streptococcus mitis Download PDFInfo
- Publication number
- CN107557317B CN107557317B CN201710862622.9A CN201710862622A CN107557317B CN 107557317 B CN107557317 B CN 107557317B CN 201710862622 A CN201710862622 A CN 201710862622A CN 107557317 B CN107557317 B CN 107557317B
- Authority
- CN
- China
- Prior art keywords
- streptococcus mitis
- product
- staphylococcus aureus
- results
- bacteriostatic
- 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
Images
Classifications
-
- 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
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention provides a streptococcus mitis and application thereof and a product using the streptococcus mitis, and relates to the technical field of microorganisms. The streptococcus mitis GDMCC60220 provided by the invention is used as probiotic bacteria, and is prepared into corresponding products which can be practically applied, so that the streptococcus mitis GDMCC60220 has great application potential in the field of medicine.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a streptococcus mitis and application thereof and a product using the streptococcus mitis.
Background
Probiotics (Probiotics) is a kind of active microorganisms beneficial to a host, and is a general term for active beneficial microorganisms which are planted in the intestinal tract and the reproductive system of a human body and can produce exact health effects so as to improve the microbial ecological balance of the host and play beneficial roles. The respiratory tract flora ecology is also a part of a human body micro-ecological system which is not neglected, and the probiotics inhabiting the respiratory tract can protect the respiratory tract from being interfered by the external environment and has certain beneficial effect on the whole body.
At present, the generation of drug-resistant bacteria caused by the abuse of antibiotics causes the reduction of the variety of the effective action of the antibiotics and the reduction of the service life, however, the probiotics effectively inhibit the growth and the reproduction of pathogenic bacteria through the probiotics or metabolites thereof, so that the problems can be well solved, and meanwhile, the probiotics also have the advantages of no toxicity, high efficiency, no residue, no drug resistance and the like.
Therefore, probiotics capable of effectively inhibiting bacteria are developed and are used for preparing corresponding products capable of being practically applied, and the application potential in the field of medicine is very huge.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The first purpose of the invention is to provide a streptococcus mitis GDMCC60220, the second purpose of the invention is to provide the application of the streptococcus mitis in preparing bacteriostatic products, and the third purpose of the invention is to provide a bacteriostatic product comprising one or more of the streptococcus mitis, fermentation broth of the streptococcus mitis or metabolite of the streptococcus mitis, so as to relieve the technical problems of serious abuse of antibiotics and blank research on probiotics in respiratory tracts in the prior art.
The invention provides streptococcus mitis with a collection number of GDMCC 60220.
Further, the streptococcus mitis has broad spectrum resistance and/or long-term antagonistic stability.
The invention also provides application of the streptococcus mitis in preparation of antibacterial products.
Further, the bacteriostasis is inhibition of one or more of staphylococcus aureus, pseudomonas aeruginosa or escherichia coli.
Further, the product is a pharmaceutical or cleaning product.
The invention also provides a bacteriostatic product, and the active component of the bacteriostatic product comprises the streptococcus mitis.
The invention also provides a bacteriostatic product, and the active component of the bacteriostatic product comprises the fermentation liquor of the streptococcus mitis.
The invention also provides a bacteriostatic product, and the active component of the bacteriostatic product comprises the metabolite of the streptococcus mitis.
Further, the bacteriostasis is inhibition of one or more of staphylococcus aureus, pseudomonas aeruginosa or escherichia coli.
Further, the product is a pharmaceutical or cleaning product.
Experiments prove that the streptococcus mitis GDMCC60220 provided by the invention has broad-spectrum resistance and long-term antagonistic stability and can play a significant role in inhibiting staphylococcus aureus, pseudomonas aeruginosa or escherichia coli. The streptococcus mitis GDMCC60220 provided by the invention is used as probiotic bacteria, and is prepared into corresponding products which can be practically applied, so that the streptococcus mitis GDMCC60220 has great application potential in the field of medicine.
Drawings
FIG. 1A is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 2-1, 2-2, 2-3, 2-4, 2-5, and 2-6 according to example 1 of the present invention;
FIG. 1B is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 2-7, 2-8, 2-9, 2-10, 2-11, and 2-12 according to example 1 of the present invention;
FIG. 1C is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 2-13, 2-14, 2-15, 2-16, 2-17, and 2-18, according to example 1 of the present invention;
FIG. 1D is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 2-19, 2-20, 2-21, 2-22, 2-23, and 2-24 according to example 1 of the present invention;
FIG. 1E is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 3-1, 3-2, 3-3, 3-4, 3-5, and 3-6 according to example 1 of the present invention;
FIG. 1F is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 3-13, 3-14, 3-15, 3-16, 3-17, and 3-18 according to example 1 of the present invention;
FIG. 1G is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 4-1, 4-2, 4-3, 4-4, 4-5, and 4-6 according to example 1 of the present invention;
FIG. 1H is a graph showing the results of screening for Staphylococcus aureus antagonism by probiotics 4-7, 4-8, 4-9, 4-10, 4-11 and 4-12 according to example 1 of the present invention;
FIG. 1I is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 4-13, 4-14, 4-15, 4-16, 4-17, and 4-18 according to example 1 of the present invention;
FIG. 1J is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 4-19, 4-20, 4-21, 4-22, 4-23, and 4-24 according to example 1 of the present invention;
FIG. 1K is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 5-1, 5-2, 5-3, 5-4, 5-5, and 5-6 according to example 1 of the present invention;
FIG. 1L is a graph showing the results of screening for Staphylococcus aureus resistance by probiotics 5-7, 5-8, 5-9, 5-10, 5-11, and 5-12 according to example 1 of the present invention;
FIG. 2 is a graph showing the results of gram staining Streptococcus mitis provided in example 1 of the present invention;
FIG. 3 is a diagram showing the results of morphological observation of Streptococcus mitis provided in example 1 of the present invention;
FIG. 4A is a graph showing the results of Escherichia coli pathogen antagonism of Streptococcus mitis after one month of culture as provided in example 2 of the present invention;
FIG. 4B is a graph showing the antagonistic effect of Streptococcus mitis on Escherichia coli pathogenic bacteria after three months of culture according to example 2 of the present invention;
FIG. 4C is a graph showing the antagonistic effect of Streptococcus mitis on Escherichia coli pathogenic bacteria after six months of culture according to example 2 of the present invention;
FIG. 4D is a graph showing the antagonistic effect of Streptococcus mitis against pathogenic bacteria of Escherichia coli after twelve months of culture according to example 2 of the present invention;
FIG. 5A is a graph showing the pathogen antagonism of Streptococcus mitis against Pseudomonas aeruginosa by Streptococcus mitis after one month of culture as provided in example 2 of the present invention;
FIG. 5B is a graph showing the pathogen antagonism of Streptococcus mitis against Pseudomonas aeruginosa after three months in culture, as provided in example 2 of the present invention;
FIG. 5C is a graph showing the pathogen antagonism of Streptococcus mitis against Pseudomonas aeruginosa after six months in culture, as provided in example 2 of the present invention;
FIG. 5D is a graph showing the pathogen antagonism of Streptococcus mitis against Pseudomonas aeruginosa after twelve months of culture as provided in example 2 of the present invention;
FIG. 6A is a graph showing the pathogen antagonistic effect of Streptococcus mitis on Staphylococcus aureus after one month of culture according to example 2 of the present invention;
FIG. 6B is a graph showing the pathogen antagonistic effect of Streptococcus mitis on Staphylococcus aureus after three-month culture according to example 2 of the present invention;
FIG. 6C is a graph showing the pathogen antagonistic effect of Streptococcus mitis on Staphylococcus aureus after six months of culture according to example 2 of the present invention;
FIG. 6D is a graph showing the pathogen antagonistic effect of Streptococcus mitis on Staphylococcus aureus after twelve months of culture according to example 2 of the present invention.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the accompanying drawings, and it should be understood that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Streptococcus mitis (Streptococcus mitis) was deposited at 7.8.2017 in Guangdong province, and the deposit unit was located at Aphance, Michelia, Mi.
In the present invention, Streptococcus mitis has broad spectrum resistance and/or long-term antagonistic stability.
The invention also provides application of the streptococcus mitis in preparation of antibacterial products.
In the present invention, the bacteriostasis is inhibition of one or more of staphylococcus aureus, pseudomonas aeruginosa or escherichia coli.
In the present invention, the product is a pharmaceutical or cleaning product.
The medicament may be, for example, but not limited to, an oral medicament or an inhalant; the cleaning product may be, for example, but not limited to, a disinfecting liquid, a washing liquid, or a soap.
In addition, the invention also provides a bacteriostatic product, and the active ingredients of the bacteriostatic product comprise any one of the following substances (a) to (c):
(a) the streptococcus mitis provided by the invention;
(b) the invention provides a fermentation liquor of streptococcus mitis;
(c) the metabolite of streptococcus mitis provided by the invention.
In the present invention, the bacteriostasis is inhibition of one or more of staphylococcus aureus, pseudomonas aeruginosa or escherichia coli.
In the present invention, the product is a pharmaceutical or cleaning product.
The medicament may be, for example, but not limited to, an oral medicament or an inhalant; the cleaning product may be, for example, but not limited to, a disinfecting liquid, a washing liquid, or a soap.
At present, the research on respiratory tract probiotics and probiotic preparations thereof is relatively few at home and abroad, and the research on humanized antagonistic probiotic strains is not found, so that the probiotic strain streptococcus mitis GDMCC60220 has broad-spectrum resistance, good antagonistic effect and good long-term antagonistic stability, and has antagonistic action on pathogenic bacteria Escherichia coli, pseudomonas aeruginosa and staphylococcus aureus.
In order to facilitate a further understanding of the present invention, the technical solutions of the present invention will now be described in detail with reference to the preferred embodiments.
Unless otherwise specified, the instruments or reagents used in the examples of the present invention are as follows:
constant temperature culture shaking table (Shanghai-Hengscientific instruments Co., Ltd.), constant temperature incubator (Shanghai Jing hong experiment equipment Co., Ltd.), sterile operating table (Sujing group Suzhou Antai air technology Co., Ltd.)
Sheep blood (solarbio), nutrient agar (Qingdao Gaokuan Haibo biotechnology, Inc.), brain heart infusion culture medium (Beijing Luoqiao technology, Inc.), serum (Hangzhou Sijiqing bioengineering materials, Inc.), broth culture medium (Beijing Obo star biotechnology, Inc.)
Example 1 isolation and identification of Streptococcus mitis
1. Isolation and culture of streptococcus mitis
Collecting pharyngeal parietal flora of 4-6 year-old healthy children, dissolving in 1mL BHI liquid culture medium, diluting the bacterial liquid by 1 × 103And (3) taking 200 mu L of bacterial liquid, coating the bacterial liquid on a 15 volume percent sheep blood culture medium, and culturing the bacterial liquid in a constant-temperature incubator at 37 ℃ for 24 hours.
The strain is different from probiotic microorganisms in human intestinal excreta in the previous research, is taken from human pharynx test, and is more convincing and reliable from the physiological position aspect on the aspect of inhibiting common bacterial infection of respiratory tract.
2. Preservation of Streptococcus mitis
Selecting a needle-point single colony with a grass green hemolytic ring by using a sterile toothpick, streaking the single colony on a sheep blood culture medium with the volume fraction of 15%, and culturing the single colony for 24 hours in a constant-temperature incubator at 37 ℃; colonies on the plates were collected and cultured overnight in 3mL of 5% fetal bovine serum BHI medium, mixed with 40% by volume of glycerol at a volume ratio of 1:1, and frozen at-80 ℃.
3. Screening of Streptococcus mitis
(1) Cultivation of pathogenic bacteria
Selecting staphylococcus aureus as pathogenic bacteria, selecting single pathogenic bacteria colony, culturing in 5mL broth culture medium, culturing overnight at 37 deg.C and 220rpm, and diluting with fresh culture solution until the concentration reaches 5-6 × 106cfu/mL for use.
(2) Streptococcus mitis antagonistic screening
Perforating a sheep blood culture medium containing probiotics with a volume fraction of 15% by using a perforator, placing a probiotic cake on a nutrient agar culture medium coated with 200 mu L of the prepared pathogenic bacteria liquid, culturing for 24 hours at 37 ℃, and observing the bacteriostasis condition, wherein the result is shown in a figure 1A-1L, and the figure 1A is the screening result of antagonistic resistance of probiotics 2-1, 2-2, 2-3, 2-4, 2-5 and 2-6 to staphylococcus aureus. FIG. 1B shows the results of screening for Staphylococcus aureus resistance by probiotics 2-7, 2-8, 2-9, 2-10, 2-11, and 2-12. FIG. 1C shows the results of screening for Staphylococcus aureus resistance by probiotics 2-13, 2-14, 2-15, 2-16, 2-17, and 2-18. FIG. 1D shows the results of screening for Staphylococcus aureus resistance by probiotics 2-19, 2-20, 2-21, 2-22, 2-23, and 2-24. FIG. 1E shows the results of screening for Staphylococcus aureus resistance by probiotics 3-1, 3-2, 3-3, 3-4, 3-5, and 3-6. FIG. 1F shows the results of screening for Staphylococcus aureus resistance by probiotics 3-13, 3-14, 3-15, 3-16, 3-17, and 3-18. FIG. 1G shows the results of screening for Staphylococcus aureus antagonistic activity against probiotic 4-1, probiotic 4-2, probiotic 4-3, probiotic 4-4, probiotic 4-5 and probiotic 4-6. FIG. 1H shows the results of screening for Staphylococcus aureus resistance by probiotics 4-7, 4-8, 4-9, 4-10, 4-11, and 4-12. FIG. 1I shows the results of screening for Staphylococcus aureus resistance by probiotics 4-13, 4-14, 4-15, 4-16, 4-17, and 4-18. FIG. 1J shows the results of screening for Staphylococcus aureus resistance by probiotics 4-19, 4-20, 4-21, 4-22, 4-23, and 4-24. FIG. 1K shows the results of screening for Staphylococcus aureus antagonistic activity against probiotic 5-1, probiotic 5-2, probiotic 5-3, probiotic 5-4, probiotic 5-5 and probiotic 5-6. FIG. 1L shows the results of screening for Staphylococcus aureus resistance by probiotics 5-7, 5-8, 5-9, 5-10, 5-11, and 5-12. As can be seen from the result graph, the probiotic 4-2 has the most significant antagonistic effect on Staphylococcus aureus, and therefore, subsequent experiments were selected.
4. Identification of Streptococcus mitis
(1)16srDNA analysis
Extracting the probiotic genome and amplifying the 16srDNA fragment according to the instructions of a gene extraction kit and a 16srDNA sequence amplification kit of Takara company.
① cells of 0.5-2.0E +09 (for Gram-positive Bacteria with a special cell wall structure and an old growth state, it is recommended that the initial amount of cells should not exceed 1.0E + 09) were collected by 1.5mL Tube, centrifuged at 12,000rpm for 2 minutes, and the supernatant (cell culture broth) was discarded.
② mu.L of Buffer BS was added to resuspend the cells, 50. mu.L of Lysozyme (20mg/mL) was added, the mixture was pipetted well and incubated in a 37 ℃ water bath for 60 minutes (the mixing could be reversed every 20 minutes during incubation).
③ 12,000,000 rpm for 5 minutes, and the supernatant was discarded.
④ mu.L of Buffer GL, 20. mu.L of protease K (20mg/mL) and 10. mu.L of RNase A (10mg/mL) were added, pipetted thoroughly and mixed, incubated in a 56 ℃ water bath for 10 minutes, at which time the solution should be clear and clear (if the solution is not clear and clear, the lysis time can be extended to 30 minutes and the mixing was performed every 5 minutes.) 200. mu.L of Buffer GB and 200. mu.L of 100% ethanol were added and mixed thoroughly.
The treated cells were performed as follows:
1. the Spin Column was mounted on a Collection Tube, the solution was transferred to the Spin Column, centrifuged at 12,000rpm for 2 minutes, and the filtrate was discarded.
2. mu.L of Buffer WA WAs added to the Spin Column, centrifuged at 12,000rpm for 1 minute, and the filtrate WAs discarded.
3. mu.L of Buffer WB was added to Spin Column, centrifuged at 12,000rpm for 1 min, and the filtrate was discarded. Note: the Buffer WB had added a specified volume of 100% ethanol. BufferWB was added around the Spin Column wall to help completely flush out salt adhering to the wall.
4. And repeating the operation step 3.
5. Spin Column was mounted on the Collection Tube and centrifuged at 12,000rpm for 2 minutes.
6. The Spin Column was placed on a new 1.5mL centrifuge tube, 50-200. mu.L of sterilized distilled water or Elution Buffer was added to the center of the Spin Column membrane, and the mixture was allowed to stand at room temperature for 5 minutes. Note: when the sterilized distilled water or the ElutionBuffer is heated to 65 ℃ for use, the elution efficiency is favorably improved.
7.12,000rpm for 2 minutes to elute the DNA. If a larger yield is required, the supernatant can be added to the center of the SpinColumn membrane again or 50-200 mu L of sterilized water or Elution Buffer is added, and after standing for 5 minutes at room temperature, the mixture is centrifuged at 12,000rpm for 2 minutes to elute DNA.
A front primer: 5'-GAGCGGATAACAATTTCACACAGG-3' (SEQ ID NO.1) and a reverse primer 5'-CGCCAGGGTTTTCCCAGTCACGAC-3' (SEQ ID NO.2), amplifying a 16srDNA fragment of probiotic 4-2, and performing sequencing by a censorship test: GCCGTGCGCGTGCTATACATGCAAGTAGAACGCTGAAGGAGGAGCTT GCTTCTCCGGATGAGTTGCGAACGGGTGAGTAACGCGTAGGTAACCT GCCTGGTAGTGGGGGATAACTATTGGAAACGATAGCTAATACCGCATA ATAGCAGTTGTTGCATGACAACTGTTTGAAAGGTGCAATTGCACCACT ACCAGATGGACCTGCGTTGTATTAGCTAGTTGGTGGGGTAACGGCTCA CCAAGGCGACGATACATAGCCGACCTGAGAGGGTGATCGGCCACACT GGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGA ATCTTCGGCAATGGACGGAAGTCTGACCGAGCAACGCCGCGTGAGTG AAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTAAGAGAAGAACGAGT GTGAGAGTGGAAAGTTCACACTGTGACGGTATCTTACCAGAAAGGGA CGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTCCCGAGCG TTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTAGATAAGT CTGAAGTTAAAGGCTGTGGCTTAACCATAGTACGCTTTGGAAACTGTT TAACTTGAGTGCAAGAGGGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTG GCTTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGAT TAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTAG ACCCTTTCCGGGGTTTAGTGCCGTAGCTAACGCATTAAGCACTCCGCC TGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGG CCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAG AACCTTACCAGGTCTTGACATCCCTCTGACCGCTCTAGAGATAGAGCT TTCCTTCGGGACAGAGGTGACAGGTGGTGCATGGTGTCGTCAGCTCG TGTCGTGAGATTGTTTGGGTTAAGTCCCGCACGAGCGCACCCCCTATG TAGTGCATCATTTAGTGGGCACTCTAGCGAGACTGCGTTATAATCGAG AAAGGTGGGGGAATGACGTCCATTCATCATGCCCGATGACTGACTACACTGCTACATGGCCTGTACACGAGCTCCGCCAGGTCGGGTGACCGAAA G (SEQ ID NO.3), identified as Streptococcus mitis by alignment with GenBank.
(2) Morphological characteristics and physiological and biochemical identification
Gram-positive gram-staining was performed on the screened Streptococcus mitis, as shown in FIG. 2. The plate colony morphology results are shown in FIG. 3. The Streptococcus mitis can be obtained by analyzing the morphology, physiological and biochemical results and combining the 16SrDNA gene with the classification position of GenBank.
The bacterial strain Streptococcus mitis is deposited in Guangdong province microorganism culture collection (GDMCC for short, address: No. 59, No. 5, Lou of Mirabilitum 100, Guangzhou city) at 8.7.2017, the deposit number is GDMCC60220, and the classification is named as Streptococcus mitis.
Example 2 bacteriostatic function verification of Streptococcus mitis
1. Cultivation of pathogenic bacteria
Respectively picking single colonies of pathogenic bacteria such as Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, culturing in 5mL broth, culturing at 37 deg.C overnight at 220rpm, and diluting the broth to 5-6 × 106cfu/mL for use.
2. Streptococcus mitis antagonism assay
(1) Enrichment of Streptococcus mitis
Selecting single colony of streptococcus mitis GDMCC60220, streaking and inoculating to sheep blood plate, culturing at 37 deg.c for 24 hr, and further enriching and amplifying pure streptococcus mitis in sheep blood plate.
(2) Determination of Streptococcus mitis antagonism and Long-term antagonism
The growth in Streptococcus mitis GDMCC60220 was scraped and vortexed thoroughly in 800. mu.L of BHI medium containing 5% fetal bovine serum for use. And respectively coating the three diluted pathogenic bacterium suspensions on nutrient agar plates, placing into an oxford cup, taking 200 mu L of streptococcus mitis in the oxford cup, culturing at 37 ℃ for 24 hours, observing the bacteriostasis condition, and measuring the bacteriostasis diameter. Streptococcus mitis GDMCC60220 was passaged once a day and antagonistic activity of Streptococcus mitis was measured at 0 month, 3 months, 6 months and 1 year, respectively. Antagonistic results against pathogenic bacteria Escherichia coli are shown in FIGS. 4A-4D, wherein FIG. 4A is the antagonistic results against pathogenic bacteria Escherichia coli by bradystreptococcus after one month of culture, FIG. 4B is the antagonistic results against pathogenic bacteria Escherichia coli by bradystreptococcus after three months of culture, FIG. 4C is the antagonistic results against pathogenic bacteria Escherichia coli by bradystreptococcus after six months of culture, and FIG. 4D is the antagonistic results against pathogenic bacteria Escherichia coli by bradystreptococcus after twelve months of culture. As can be seen from the result graph, the antibacterial effect on the pathogenic bacteria of the Escherichia coli is not changed in different passages, namely 0, 3, 6 and 12 months, and the Escherichia coli still has good antagonistic property after passage for 12 months, wherein the antibacterial diameter is about 19.5 mm. The antagonistic results against pseudomonas aeruginosa are shown in fig. 5A-5D, wherein fig. 5A is the antagonistic result against pseudomonas aeruginosa pathogenic bacteria by streptococcus mitis after one month of culture, fig. 5B is the antagonistic result against pseudomonas aeruginosa pathogenic bacteria by streptococcus mitis after three months of culture, fig. 5C is the antagonistic result against pseudomonas aeruginosa pathogenic bacteria by streptococcus mitis after six months of culture, and fig. 5D is the antagonistic result against pseudomonas aeruginosa pathogenic bacteria by streptococcus mitis after twelve months of culture. As can be seen from the result graph, the bacteriostatic effect on the pseudomonas aeruginosa pathogenic bacteria is not changed in different passages, namely 0 month, 3 months, 6 months and 12 months, and the pseudomonas aeruginosa pathogenic bacteria still have good antagonistic property after being passed for 12 months, wherein the bacteriostatic diameter is about 19.0 mm. The results of antagonism against staphylococcus aureus are shown in fig. 6A-6D, where fig. 6A is the results of antagonism against staphylococcus aureus pathogenic bacteria by bradystreptococcus after one month of culture, fig. 6B is the results of antagonism against staphylococcus aureus pathogenic bacteria by bradystreptococcus after three months of culture, fig. 6C is the results of antagonism against staphylococcus aureus pathogenic bacteria by bradystreptococcus after six months of culture, and fig. 6D is the results of antagonism against staphylococcus aureus pathogenic bacteria by bradystreptococcus after twelve months of culture. As can be seen from the result graph, the bacteriostatic effect on staphylococcus aureus pathogenic bacteria is not changed in different passages, namely 0, 3, 6 and 12 months, and the staphylococcus aureus pathogenic bacteria still have good antagonism after being subjected to passage for 12 months, wherein the bacteriostatic diameter is about 21.0 mm.
In conclusion, experiments prove that the streptococcus mitis GDMCC60220 provided by the invention has broad-spectrum resistance and long-term antagonistic stability, and can play a significant role in inhibiting staphylococcus aureus, pseudomonas aeruginosa or escherichia coli. The streptococcus mitis GDMCC60220 provided by the invention is used as probiotic bacteria, and is prepared into corresponding products which can be practically applied, so that the streptococcus mitis GDMCC60220 has great application potential in the field of medicine.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
SEQUENCE LISTING
<110> Shenyang medical college
<120> Streptococcus mitis, use thereof and products using the same
<160>3
<170>PatentIn version 3.5
<210>1
<211>24
<212>DNA
<213> Artificial sequence
<400>1
gagcggataa caatttcaca cagg 24
<210>2
<211>24
<212>DNA
<213> Artificial sequence
<400>2
cgccagggtt ttcccagtca cgac 24
<210>3
<211>1232
<212>DNA
<213> Streptococcus mitis
<400>3
gccgtgcgcg tgctatacat gcaagtagaa cgctgaagga ggagcttgct tctccggatg 60
agttgcgaac gggtgagtaa cgcgtaggta acctgcctgg tagtggggga taactattgg 120
aaacgatagc taataccgca taatagcagt tgttgcatga caactgtttg aaaggtgcaa 180
ttgcaccact accagatgga cctgcgttgt attagctagt tggtggggta acggctcacc 240
aaggcgacga tacatagccg acctgagagg gtgatcggcc acactgggac tgagacacgg 300
cccagactcc tacgggaggc agcagtaggg aatcttcggc aatggacgga agtctgaccg 360
agcaacgccg cgtgagtgaa gaaggttttc ggatcgtaaa gctctgttgt aagagaagaa 420
cgagtgtgag agtggaaagt tcacactgtg acggtatctt accagaaagg gacggctaac 480
tacgtgccag cagccgcggt aatacgtagg tcccgagcgt tgtccggatt tattgggcgt 540
aaagcgagcg caggcggtta gataagtctg aagttaaagg ctgtggctta accatagtac 600
gctttggaaa ctgtttaact tgagtgcaag aggggagagt ggaattccat gtgtagcggt 660
gaaatgcgta gatatatgga ggaacaccgg tggcgaaagc ggctctctgg cttgtaactg 720
acgctgaggc tcgaaagcgt ggggagcaaa caggattaga taccctggta gtccacgccg 780
taaacgatga gtgctaggtg ttagaccctt tccggggttt agtgccgtag ctaacgcatt 840
aagcactccg cctggggagt acgaccgcaa ggttgaaact caaaggaatt gacgggggcc 900
cgcacaagcg gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaggtct 960
tgacatccct ctgaccgctc tagagataga gctttccttc gggacagagg tgacaggtgg 1020
tgcatggtgt cgtcagctcg tgtcgtgaga ttgtttgggt taagtcccgc acgagcgcac 1080
cccctatgta gtgcatcatt tagtgggcac tctagcgaga ctgcgttata atcgagaaag 1140
gtgggggaat gacgtccatt catcatgccc gatgactgac tacactgcta catggcctgt 1200
acacgagctc cgccaggtcg ggtgaccgaa ag 1232
Claims (7)
1. The application of streptococcus mitis in preparing bacteriostatic products;
the collection number of streptococcus mitis is GDMCC 60220.
2. The use according to claim 1, wherein the bacteriostatic action is inhibition of one or more of staphylococcus aureus, pseudomonas aeruginosa or escherichia coli.
3. Use according to claim 1 or 2, wherein the product is a pharmaceutical or cleaning product.
4. A bacteriostatic product, wherein the active ingredient of the bacteriostatic product comprises streptococcus mitis as claimed in claim 1.
5. A bacteriostatic product, wherein the active ingredient of the bacteriostatic product comprises the fermentation broth of Streptococcus mitis of claim 1.
6. The product of any one of claims 4-5, wherein the bacteriostatic agent is one or more of Staphylococcus aureus, Pseudomonas aeruginosa, or Escherichia coli.
7. A product according to any of claims 4 to 5, wherein the product is a pharmaceutical or cleaning product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710862622.9A CN107557317B (en) | 2017-09-20 | 2017-09-20 | Streptococcus mitis, application thereof and product using streptococcus mitis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710862622.9A CN107557317B (en) | 2017-09-20 | 2017-09-20 | Streptococcus mitis, application thereof and product using streptococcus mitis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107557317A CN107557317A (en) | 2018-01-09 |
| CN107557317B true CN107557317B (en) | 2020-03-24 |
Family
ID=60982585
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710862622.9A Active CN107557317B (en) | 2017-09-20 | 2017-09-20 | Streptococcus mitis, application thereof and product using streptococcus mitis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107557317B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110904010B (en) * | 2019-12-17 | 2021-11-02 | 南方医科大学 | Arthrobacter phenantivorans L131 strain for antagonizing respiratory pathogenic bacteria and application thereof |
| CN111100821B (en) * | 2020-01-16 | 2023-07-14 | 沈阳医学院 | Streptococcus and application thereof |
| CN116042432B (en) * | 2022-07-14 | 2024-07-23 | 沈阳医学院 | Novel strain with broad-spectrum antibacterial property and purification of antibacterial substances thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103119153A (en) * | 2010-08-31 | 2013-05-22 | 公共健康高级研究中心 | Anticaries compositions and probiotics/prebiotics |
-
2017
- 2017-09-20 CN CN201710862622.9A patent/CN107557317B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103119153A (en) * | 2010-08-31 | 2013-05-22 | 公共健康高级研究中心 | Anticaries compositions and probiotics/prebiotics |
Non-Patent Citations (1)
| Title |
|---|
| 饲用抑菌活性乳酸菌共培养筛选、优化及抗性分析;徐秀伟;《中国优秀硕士学位论文全文数据库 农业科技辑》;20170515(第5（2017）期);正文第11页第1.2.1.1小节，第16页第2.1.1小节，表2-1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107557317A (en) | 2018-01-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107460150B (en) | Dentisiana streptococcus, application thereof and product using same | |
| Abdulkadir et al. | Screening and isolation of the soil bacteria for ability to produce antibiotics | |
| CN107557317B (en) | Streptococcus mitis, application thereof and product using streptococcus mitis | |
| US10221440B2 (en) | Method for determining the sensitivity of microorganisms to antimicrobial substances | |
| Macià et al. | Microbiological diagnosis of biofilm-related infections | |
| CN101120084A (en) | Methods and compositions for ultra-high throughput screening of natural products | |
| Bharti et al. | Heat stable antimicrobial activity of Burkholderia gladioli OR1 against clinical drug resistant isolates | |
| Lochhead et al. | Aspects of antagonisms between micro-organisms in soil | |
| CN116410897A (en) | Friedson escherichia and antibacterial application thereof | |
| CN114246868B (en) | Inhibitor of urinary tract infection pathogen urinary tract pathogenic escherichia coli | |
| Packialakshmi et al. | Role of effective microorganism in unfertile soil | |
| Al-Hamadany | Biofilm formation in MRSA Staphylococcus aureus isolated from diabetics' UTI | |
| Ukhty et al. | Isolation of endophytic fungi from the coastal plant terong pungo (Solanum sp.) and its antibacterial activity against oral pathogenic bacteria | |
| Al-Kabi et al. | Detection of Pseudomonas aeruginosa with Biofilm Formation in Burn Patients: A Study in Al-Hussein Teaching Hospital of Iraq | |
| RIZQOH et al. | Exploring the antibacterial activity of endophytic bacteria from Andaliman (Zanthoxylum acanthopodium DC.) against Bacillus subtilis | |
| CN118389335A (en) | Streptococcus with antibacterial effect and application thereof | |
| Hadke et al. | Anti-Microbial Activity of Antibiotic Produced From Fungi by Fermentation Process. | |
| CN116042432B (en) | Novel strain with broad-spectrum antibacterial property and purification of antibacterial substances thereof | |
| Seal et al. | Phenotypic and Molecular Characterizations of Haemolytic and Penicillin-Resistant Bacillus cereus and its Control by Plant Extracts | |
| CN118667685A (en) | Bacterial strain separated from healthy children respiratory tract pharyngeal swab and application thereof | |
| Azode et al. | Screening of antibiotic-producing bacteria isolated from soil gotten within gaba, Bwari FCT, Abuja | |
| Sartika | Screening of Endophytic Bacteria from Purwoceng Roots (Pimpinella pruatjan Molk.) Antibacterial Activity Against Staphylococcus aureus | |
| DARMESTI et al. | IDENTIFICATION OF ANTIBIOTIC RESISTANCE PATTERNS IN Escherichia coli BACTERIA FROM CLOACAL SWAB SAMPLES OF BROILER CHICKENS FROM FARM THAT USE PROBIOTIC Lactobacillus sp. | |
| US20110275114A1 (en) | Method and medium for detecting the presence or absence of methicillin resistant staphylococcus aureus (mrsa) in a test sample | |
| Parra et al. | Unknown Soil Isolates from Urban Park as Antibiotic Producers |
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 |