CN115093989A - Streptococcus salivarius QS-20 with antioxidant activity and application thereof - Google Patents
Streptococcus salivarius QS-20 with antioxidant activity and application thereof Download PDFInfo
- Publication number
- CN115093989A CN115093989A CN202210562002.4A CN202210562002A CN115093989A CN 115093989 A CN115093989 A CN 115093989A CN 202210562002 A CN202210562002 A CN 202210562002A CN 115093989 A CN115093989 A CN 115093989A
- Authority
- CN
- China
- Prior art keywords
- streptococcus salivarius
- strain
- antioxidant
- preparation
- streptococcus
- 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.)
- Granted
Links
- 241000194024 Streptococcus salivarius Species 0.000 title claims abstract description 49
- 230000003078 antioxidant effect Effects 0.000 title claims abstract description 22
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 claims abstract description 11
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 10
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 7
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 7
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims abstract description 7
- 241000588724 Escherichia coli Species 0.000 claims abstract description 6
- 241000607142 Salmonella Species 0.000 claims abstract description 6
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 6
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 6
- 238000004321 preservation Methods 0.000 claims abstract description 6
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 238000002360 preparation method Methods 0.000 claims description 8
- 239000003963 antioxidant agent Substances 0.000 claims description 7
- 239000000022 bacteriostatic agent Substances 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 238000000855 fermentation Methods 0.000 claims description 3
- 230000004151 fermentation Effects 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 230000009471 action Effects 0.000 claims description 2
- 239000002773 nucleotide Substances 0.000 claims description 2
- 125000003729 nucleotide group Chemical group 0.000 claims description 2
- 239000007858 starting material Substances 0.000 claims 3
- 230000002000 scavenging effect Effects 0.000 claims 1
- 239000002253 acid Substances 0.000 abstract description 9
- 230000003647 oxidation Effects 0.000 abstract description 5
- 238000007254 oxidation reaction Methods 0.000 abstract description 5
- 241000894006 Bacteria Species 0.000 abstract description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 4
- 244000052616 bacterial pathogen Species 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 210000000941 bile Anatomy 0.000 abstract 1
- 230000002349 favourable effect Effects 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 description 11
- 239000003833 bile salt Substances 0.000 description 8
- 210000004251 human milk Anatomy 0.000 description 7
- 235000020256 human milk Nutrition 0.000 description 7
- 238000009630 liquid culture Methods 0.000 description 7
- 230000012010 growth Effects 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- 241000194017 Streptococcus Species 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 241000192125 Firmicutes Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 230000003698 anagen phase Effects 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 235000010855 food raising agent Nutrition 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 241000186000 Bifidobacterium Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 241000191940 Staphylococcus Species 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000002856 computational phylogenetic analysis Methods 0.000 description 2
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 229940039696 lactobacillus Drugs 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- KEQXNNJHMWSZHK-UHFFFAOYSA-L 1,3,2,4$l^{2}-dioxathiaplumbetane 2,2-dioxide Chemical compound [Pb+2].[O-]S([O-])(=O)=O KEQXNNJHMWSZHK-UHFFFAOYSA-L 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- -1 DPPH free radical Chemical class 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 241000192132 Leuconostoc Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000002390 adhesive tape Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- 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
-
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/245—Salivarius
-
- 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)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Public Health (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Epidemiology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Molecular Biology (AREA)
- Toxicology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a streptococcus salivarius QS-20 with antioxidant activity and application thereof. The classification name of the streptococcus salivarius QS-20 isStreptococcus salivariusThe strain is preserved in China general microbiological culture Collection center with the preservation number of CGMCC 7.456, and can effectively remove oxidation substances such as DPPH free radicals, hydrogen peroxide and the like; also has good acid-producing and acid-resisting properties and bile salt-resisting capability, is favorable for the permanent planting of the bacteria in intestinal tracts, andit has excellent inhibitory activity against various pathogenic bacteria such as Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Salmonella and Pseudomonas aeruginosa. Therefore, QS-20 of streptococcus salivarius has wide application scenes.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to streptococcus salivarius QS-20 with antioxidant activity and application thereof.
Background
Human milk is rich in microorganisms (including bacteria, viruses and fungi) which play an important role not only in the initiation of the neonatal gut immune system, the development of the infant gut immune function and the programmed process, but also in protecting the infant from diseases and infections during the growth process. At present, microorganisms which have been isolated and identified from breast milk are mainly staphylococcus, streptococcus, enterococcus, lactobacillus, bifidobacterium, leuconostoc and the like. Wherein, staphylococcus, streptococcus, bifidobacterium and lactobacillus are the dominant flora in the breast milk and have important probiotic function. Therefore, the research and development of the human milk streptococcus have important significance.
Disclosure of Invention
The invention aims to provide a streptococcus salivarius QS-20 with antioxidant activity and application thereof, wherein the streptococcus salivarius QS-20 has antioxidant activity, acid production characteristic, bile salt resistance and bacteriostatic ability.
In order to realize the purpose of the invention, the invention adopts the following technical scheme to realize:
the invention provides streptococcus salivarius QS-20 with antioxidant activity, and the classification of the streptococcus salivarius QS-20 isStreptococcus salivariusThe culture medium is preserved in China general microbiological culture Collection center, and the address is as follows: xilu No.1 Hospital No. 3, Chaojing, Chaoyang, with a preservation number of CGMCC 7.456, and a preservation date: 13 months 1 in 2022; the result of the viability test of the biomaterial is as follows: and (6) survival.
Furthermore, the streptococcus salivarius QS-20 is a white protruding colony, the surface is smooth, the diameter of the colony is 0.2-1mm, cells are spherical or oval, the diameter is not more than 1 mu m, and the cells are arranged in a chain shape.
Furthermore, the streptococcus salivarius QS-20 has antioxidant activity, acid production property, bile salt resistance and bacteriostatic ability.
Further, the nucleotide sequence of QS-20 of streptococcus salivarius is shown as SEQ ID No.1
The invention also provides application of QS-20 in streptococcus salivarius in preparing an antioxidant.
Further, the action time of the antioxidant is 12-36 h.
Furthermore, the streptococcus salivarius QS-20 achieves the effect of resisting oxidation by eliminating DPPH free radicals and hydrogen peroxide.
The invention also provides application of the streptococcus salivarius QS-20 in preparation of a leavening agent.
Further, the fermentation time of the leavening agent is 0-3 h.
The invention also provides application of the streptococcus salivarius QS-20 in preparation of a bacteriostatic agent.
Further, the bacteriostatic agent can inhibit escherichia coli, bacillus subtilis, staphylococcus aureus, salmonella and pseudomonas aeruginosa.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the probiotic is separated from the breast milk, and the streptococcus salivarius QS-20 with good oxidation resistance is obtained by taking DPPH clearance as a screening index, has good oxidation resistance activity, and can effectively remove oxidation substances such as DPPH free radicals and hydrogen peroxide; also has good acid-producing and acid-resisting properties, and provides basis for the application of the compound in fermented foods. In addition, the streptococcus salivarius QS-20 also has bile salt resistance, is beneficial to colonization in intestinal tracts and has good inhibitory activity on various pathogenic bacteria such as escherichia coli, bacillus subtilis, staphylococcus aureus, salmonella and pseudomonas aeruginosa. Therefore, QS-20 of streptococcus salivarius has wide application scenes.
Drawings
FIG. 1 is an electrophoretogram of 16S rDNA amplification PCR products of a part of strains.
FIG. 2 is a QS-20 strain phylogenetic tree analysis.
FIG. 3 is a colony morphology and a cell morphology map of QS-20 of Streptococcus salivarius.
FIG. 4 shows the growth and acidogenesis curves of QS-20 of Streptococcus salivarius.
FIG. 5 is a graph showing DPPH free radical scavenging effect of QS-20 in Streptococcus salivarius.
FIG. 6 is an acid resistance curve for Streptococcus salivarius QS-20.
FIG. 7 is a bile salt resistance curve for S.salivarius QS-20.
FIG. 8 is a graph showing the inhibitory effect of QS-20 of Streptococcus salivarius on bacteria.
Detailed Description
The technical solution of the present invention will be further described in detail with reference to the following specific examples.
In the following examples, unless otherwise specified, the experimental methods used were all conventional methods, and materials, reagents and the like used were all available from biological or chemical reagents companies.
Example 1: isolation screening and characterization of QS-20 of Streptococcus salivarius
1. Isolation and identification
Fresh breast milk is diluted by physiological saline according to different concentrations and coated on MRS solid culture medium, and is subjected to inverted culture at 37 ℃ for 48 h. Selecting a single bacterial colony with smooth and white or milk white thallus for gram staining and microscopic examination, selecting rod-shaped and gram-positive bacteria, and streaking and purifying for 2-3 times on an MRS solid culture medium. And selecting a streaked and purified single colony, inoculating the single colony to an MRS liquid culture medium, continuously culturing for 24-48 h at 37 ℃, mixing bacterial liquid and sterilized glycerol according to a ratio of 1: 1, and storing in a refrigerator at-80 ℃ for later use.
(1) Extracting genome DNA by adopting a radix asparagi genome kit;
(2) 16S rDNA PCR amplification, adopting universal primer,
the upstream primer 27F: GAGAGTTTGATCCTGGCTCAG, and the adhesive tape is used for adhering the film to a substrate,
the downstream primer 1492R: TACGGCTACCTTGTTACGAC are provided.
The PCR reaction system is 50 ul: taq Reaction Buffer (10X) 5ul, PCR Dye 5ul, dNTPs 1ul, Taq Dymerase 0.5ul, DNA template 2ul, 27F 2ul, 1492R 2ul, water to make up to 50 ul.
The PCR amplification procedure was: pre-denaturation at 95 ℃ for 5 min; circulating for 30 times (denaturation at 95 deg.C for 30 s; annealing at 55 deg.C for 30 s; extension at 72 deg.C for 90 s); extension at 72 ℃ for 7min, and extension at the end for 20 ℃ for 3 min.
(3) Sequencing of PCR amplification products
The PCR product was detected by 1% agarose gel electrophoresis and sent to Qingdao Biotech for sequencing. The sequences were determined to be aligned for homology with the GenBank database by the BLAST program.
16 bacteria are separated from three different breast milk samples by a transparent circle method, 16S rDNA amplification is carried out to obtain target bands (figure 1) of about 1500 bp, and the nucleotide sequences are shown as SEQ ID No. 1.
Sequencing the target band and analyzing phylogenetic tree, and obtaining QS-20 strain and QS-20 strain from the phylogenetic tree shown in FIG. 2Streptococcus salivariusThe relationship is recent and therefore it belongs to the streptococcus salivarius.
Strain QS-20 is deposited, the unit of deposit of streptococcus salivarius QS-20: china general Committee for culture Collection of microorganismsMicrobial center (CGMCC); address: west road No.1, north zhou zhao yang district, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: 1 month and 13 days 2022; the classification is named as: salivary streptococcusStreptococcus salivarius，The preservation number is CGMCC 7.456, and the detection result of the viability of the biological material is as follows: and (6) survival.
2. Morphological observations
The QS-20 strain was observed for colony morphology and gram-stained cell morphology, as shown in FIG. 3. QS-20 strain white convex colony, smooth surface, 0.2-1mm diameter, spherical or oval cell with diameter not more than 1 μm, chain arrangement, and precipitation growth in liquid culture medium. The QS-20 strain can produce acid to form a transparent ring on an MRS culture medium added with calcium carbonate, the gram staining result shows that the QS-20 strain is gram positive bacteria, meanwhile, the catalase test of the strain is negative, and the result further shows that the QS-20 strain is streptococcus in the gram positive bacteria, which is consistent with the result of the phylogenetic tree analysis.
Example 2: biochemical Properties of Streptococcus salivarius QS-20
1. Growth and acid production curves of the QS-20 Strain of Streptococcus salivarius
Referring to fig. 4, the QS-20 strain can rapidly enter the logarithmic growth phase within a short time, has strong adaptability and short adaptation period, enters the exponential growth phase within 5-15h and then enters the stationary phase, and has no obvious decay phase within 52 h. In the exponential growth phase, the acid-producing property of streptococcus salivarius QS-20 is strong, and the pH value is obviously reduced, so that the streptococcus salivarius QS-20 is suitable for serving as a leavening agent.
2. Determination of antioxidant Activity of Streptococcus salivarius QS-20 Strain
The DPPH free radical is reduced by the antioxidant and becomes stable DPPH molecule, the color is changed from dark purple to light yellow, and the purple intensity of DPPH is inversely proportional to the antioxidant activity of the sample. DPPH has a stable structure, and if DPPH can be eliminated, the test strain has the chain reaction effects of reducing hydrogen peroxide and eliminating various free radicals. The QS-20 strain of streptococcus salivarius is inoculated in MRS broth liquid culture medium, and the antioxidant activity of fermentation supernatant is measured at different culture time.
As shown in FIG. 5, it can be seen that QS-20 strain has high DPPH clearance rate, which reaches more than 70% after 12h and more than 80% after 36h, and shows strong antioxidant activity.
3. Determination of acid resistance of QS-20 Strain of Streptococcus salivarius
QS-20 strain is inoculated into MRS broth liquid culture medium with pH 3.0 for culturing for 3 and 5 hours, the liquid culture medium without thalli and with pH 3.0 is used as a blank, OD600nm is measured, the culture time is used as abscissa, the absorbance value is used as slave coordinate, and a broken line trend graph is drawn.
As can be seen in FIG. 6, the absorbance gradually increased within 3h, indicating that QS-20 strain can survive and grow; the absorbance at 5h was not changed much compared to 3h, indicating that the growth of the strain was inhibited. Therefore, the maximum survival tolerance time of the strain under the acidic condition is 0-3 h.
4. Determination of bile salt resistance of QS-20 strain of streptococcus salivarius
QS-20 strain is inoculated into MRS broth liquid culture medium containing different concentrations of bile salt (0.1%, 0.2%, 0.3%) and cultured for 3 and 5 hours, the liquid culture medium containing no thalli and different concentrations of bile salt is used as a blank, the absorbance of OD600nm is measured, the culture time is used as an abscissa, the absorbance value is used as an ordinate, and a broken line trend graph is drawn.
As shown in FIG. 7, QS-20 of streptococcus salivarius can survive and grow within 5h, and after 3h, the high concentration of bile salt can inhibit the growth of the strain to some extent.
Example 3: analysis of bacteriostatic Capacity of Streptococcus salivarius QS-20
Culturing Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Salmonella and Pseudomonas aeruginosa, and detecting the antibacterial activity of Streptococcus salivarius QS-20 on the strains.
The results in FIG. 8 show that the QS-20 strains of Streptococcus salivarius have inhibitory activity against Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Salmonella, and Pseudomonas aeruginosa, among which the inhibitory activity against Bacillus subtilis and Pseudomonas aeruginosa is stronger.
The above embodiments are only used to illustrate the technical solution of the present invention, and not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing embodiments, or equivalents may be substituted for some of the features thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Sequence listing
<110> Qingdao agricultural university
<120> streptococcus salivarius QS-20 with antioxidant activity and application thereof
<141> 2022-04-08
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1447
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
gggcggggtg tctttactgc agtagaacgc tgaagagagg agcttgctct tcttggatga 60
gttgcgaacg ggtgagtaac gcgtaggtaa cctgccttgt agcgggggat aactattgga 120
aacgatagct aataccgcat aacaatggat gacacatgtc atttatttga aaggggcaat 180
tgctccacta caagatggac ctgcgttgta ttagctagta ggtgaggtaa cggctcacct 240
aggcgacgat acatagccga cctgagaggg tgatcggcca cactgggact gagacacggc 300
ccagactcct acgggaggca gcagtaggga atcttcggca atgggggcaa ccctgaccga 360
gcaacgccgc gtgagtgaag aaggttttcg gatcgtaaag ctctgttgta agtcaagaac 420
gagtgtgaga gtggaaagtt cacactgtga cggtagctta ccagaaaggg acggctaact 480
acgtgccagc agccgcggta atacgtaggt cccgagcgtt gtccggattt attgggcgta 540
aagcgagcgc aggcggtttg ataagtctga agttaaaggc tgtggctcaa ccatagttcg 600
ctttggaaac tgtcaaactt gagtgcagaa ggggagagtg gaattccatg tgtagcggtg 660
aaatgcgtag atatatggag gaacaccggt ggcgaaagcg gctctctggt ctgtaactga 720
cgctgaggct cgaaagcgtg gggagcgaac aggattagat accctggtag tccacgccgt 780
aaacgatgag tgctaggtgt tggatccttt tcgggattca gtgccgcagc taacgcatta 840
agcactccgc ctggggagta cgaccgcaag gttgaaactc aaaggaattg acgggggccc 900
gcacaagcgg tggagcatgt ggtttaattc gaagcaacgc gaagaacctt accaggtctt 960
gacatcccga tgctatttct agagatagaa agttacttcg gtacatcggt gacaggtggt 1020
gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac 1080
ccctattgtt agttgccatc attcagttgg gcactctagc gagactgccg gtaataaacc 1140
ggaggaaggt ggggatgacg tcaaatcatc atgcccctta tgacctgggc tacacacgtg 1200
ctacaatggt tggtacaacg agttgcgagt cggtgacggc aagctaatct cttaaagcca 1260
atctcagttc ggattgtagg ctgcaactcg cctacatgaa gtcggaatcg ctagtaatcg 1320
cggatcagca cgccgcggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca 1380
cgagagtttg taacacccga agtcggtgag gtaacctttg gagccagccg cctaaggtca 1440
agatgtt 1447
Claims (10)
1. A strain of streptococcus salivarius QS-20 with antioxidant activity, wherein the classification of the streptococcus salivarius QS-20 is shown asStreptococcus salivariusThe strain is preserved in China general microbiological culture Collection center with the preservation number of CGMCC 7.456.
2. The process according to claim 1, wherein said QS-20 streptococcus salivarius are white protruding colonies with smooth surface, 0.2-1mm diameter colonies, spherical or oval cells with diameter not more than 1 μm, and chain-like arrangement.
3. The method according to claim 1, wherein the nucleotide sequence of QS-20 is SEQ ID No. 1.
4. Use of a streptococcus salivarius QS-20 as claimed in any of claims 1 to 3 in the preparation of an antioxidant.
5. The use of QS-20 in the preparation of an antioxidant according to claim 4, wherein the antioxidant has an action time of 12-36 hours.
6. Use of QS-20 in the preparation of an antioxidant according to claim 4, wherein the QS-20 has antioxidant activity by scavenging DPPH free radicals and hydrogen peroxide.
7. Use of a streptococcus salivarius QS-20 as claimed in any of claims 1 to 3 in the preparation of a starter culture.
8. The use of QS-20 in accordance with claim 6, for the preparation of a starter culture, wherein the starter culture has a fermentation time of 0-3 h.
9. Use of a streptococcus salivarius QS-20 as claimed in any of claims 1 to 3 in the manufacture of a bacteriostatic agent.
10. Use of QS-20 to streptococcus salivarius for the preparation of a bacteriostatic agent according to claim 9 wherein the bacteriostatic agent is capable of inhibiting escherichia coli, bacillus subtilis, staphylococcus aureus, salmonella and pseudomonas aeruginosa.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2022103636278 | 2022-04-08 | ||
| CN202210363627 | 2022-04-08 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115093989A true CN115093989A (en) | 2022-09-23 |
| CN115093989B CN115093989B (en) | 2023-06-06 |
Family
ID=83288671
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210562002.4A Active CN115093989B (en) | 2022-04-08 | 2022-05-23 | Streptococcus salivarius QS-20 with antioxidant activity and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115093989B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116574634A (en) * | 2023-03-13 | 2023-08-11 | 广东悦创生物科技有限公司 | Streptococcus salivarius thermophilus subspecies JF2 and application thereof in preparation of anti-inflammatory and lipid-relieving food and drug |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5468479A (en) * | 1991-06-28 | 1995-11-21 | Toshio Kawanishi | Compositions containing lactic acid bacterium Streptococcus salivarius |
| BRPI0605568A (en) * | 2006-12-06 | 2008-07-22 | Vera Fantinato | composition, food product, fermented milk, fermentation milk preparation process and use of streptococcus salivarius subsp salivarius bacteria |
| CN101538545A (en) * | 2009-04-08 | 2009-09-23 | 内蒙古蒙牛乳业（集团）股份有限公司 | Lactobacillus salivarius and food composition thereof |
| RU2567148C1 (en) * | 2014-12-24 | 2015-11-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Горский государственный аграрный университет" | Streptococcus salivarius VKPM V-11177 STRAIN USED FOR PRODUCTION OF FERMENTED MILK PRODUCTS AND PRO-BIOTIC PREPARATIONS |
| RU2698032C1 (en) * | 2018-11-08 | 2019-08-21 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Горский государственный аграрный университет" | Strain of lactobacilli streptococcus salivarius - producer of lactic acid and antibiotic substances |
| KR20200002008A (en) * | 2018-06-28 | 2020-01-07 | 주식회사 세바바이오텍 | Streptococcus salivarius sbk4 strain isolated from skin and functional cosmetic composition comprising Streptococcus salivarius sbk4 strain |
| JP6651202B1 (en) * | 2018-10-17 | 2020-02-19 | 株式会社北岡本店 | A novel lactic acid bacterium belonging to Streptococcus salivarius and its use |
| WO2022049244A1 (en) * | 2020-09-04 | 2022-03-10 | Universiteit Antwerpen | A novel probiotic streptococcus salivarius strain and its uses |
-
2022
- 2022-05-23 CN CN202210562002.4A patent/CN115093989B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5468479A (en) * | 1991-06-28 | 1995-11-21 | Toshio Kawanishi | Compositions containing lactic acid bacterium Streptococcus salivarius |
| BRPI0605568A (en) * | 2006-12-06 | 2008-07-22 | Vera Fantinato | composition, food product, fermented milk, fermentation milk preparation process and use of streptococcus salivarius subsp salivarius bacteria |
| CN101538545A (en) * | 2009-04-08 | 2009-09-23 | 内蒙古蒙牛乳业（集团）股份有限公司 | Lactobacillus salivarius and food composition thereof |
| RU2567148C1 (en) * | 2014-12-24 | 2015-11-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Горский государственный аграрный университет" | Streptococcus salivarius VKPM V-11177 STRAIN USED FOR PRODUCTION OF FERMENTED MILK PRODUCTS AND PRO-BIOTIC PREPARATIONS |
| KR20200002008A (en) * | 2018-06-28 | 2020-01-07 | 주식회사 세바바이오텍 | Streptococcus salivarius sbk4 strain isolated from skin and functional cosmetic composition comprising Streptococcus salivarius sbk4 strain |
| JP6651202B1 (en) * | 2018-10-17 | 2020-02-19 | 株式会社北岡本店 | A novel lactic acid bacterium belonging to Streptococcus salivarius and its use |
| RU2698032C1 (en) * | 2018-11-08 | 2019-08-21 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Горский государственный аграрный университет" | Strain of lactobacilli streptococcus salivarius - producer of lactic acid and antibiotic substances |
| WO2022049244A1 (en) * | 2020-09-04 | 2022-03-10 | Universiteit Antwerpen | A novel probiotic streptococcus salivarius strain and its uses |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116574634A (en) * | 2023-03-13 | 2023-08-11 | 广东悦创生物科技有限公司 | Streptococcus salivarius thermophilus subspecies JF2 and application thereof in preparation of anti-inflammatory and lipid-relieving food and drug |
| CN116574634B (en) * | 2023-03-13 | 2023-11-03 | 广东悦创生物科技有限公司 | Streptococcus salivarius thermophilus subspecies JF2 and application thereof in preparation of anti-inflammatory and lipid-relieving food and drug |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115093989B (en) | 2023-06-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108148789B (en) | Lactobacillus rhamnosus and application thereof in preparation of bacteriocin | |
| CN110157645B (en) | Lactobacillus salivarius Y4 and application thereof | |
| CN108611299B (en) | Lactobacillus plantarum for producing antibacterial peptide and application thereof | |
| CN111534452B (en) | Lactobacillus strain with broad-spectrum inhibition of multi-drug-resistance food-borne pathogenic bacteria and application thereof | |
| CN114196593B (en) | Lactobacillus plantarum P101 with high antioxidant activity and application thereof | |
| CN110878270B (en) | Lactobacillus paracasei subspecies paracasei and application thereof | |
| CN113846039A (en) | Bacillus belgii and application thereof | |
| CN115093989B (en) | Streptococcus salivarius QS-20 with antioxidant activity and application thereof | |
| CN113980853B (en) | Lactic acid-producing lactococcus garvieae WBT0008 and application thereof | |
| CN108004177B (en) | Lactobacillus paracasei capable of degrading nitrite and characteristic research thereof | |
| CN109897800B (en) | Selenium-rich enterococcus durans A8-1 and application thereof | |
| CN116478873B (en) | Bacillus bailii capable of inhibiting growth of various harmful bacteria in intestinal tracts of Min pigs | |
| CN110106113B (en) | Lactobacillus kefir MSR101 and application thereof | |
| CN114717150B (en) | Lactobacillus plantarum CRS33 and application thereof | |
| CN116590172A (en) | Pediococcus acidilactici GLP06, application and product thereof | |
| CN106591176B (en) | Lactobacillus pentosus and application thereof | |
| CN113717887B (en) | Goose-source lactobacillus plantarum and application thereof | |
| CN113061550B (en) | Lactobacillus new strain Z6 and application thereof in food | |
| CN113604387B (en) | Salt-tolerant and high-temperature-resistant lactobacillus reuteri and application thereof in prevention and treatment of pathogenic bacteria in livestock and poultry aquaculture | |
| CN111363703B (en) | Weissella strain with antibacterial and antioxidant activities and application thereof | |
| CN115322921A (en) | Bacillus having antagonistic action on pathogenic bacteria of cow mastitis and application thereof | |
| CN113215067A (en) | VBNC (viable but non-viable) state lactobacillus brevis CSHRR5-3 strain and application thereof | |
| KR101972494B1 (en) | Noverl microalgae having resistance against selenium | |
| CN113355265B (en) | Lactobacillus plantarum and application thereof | |
| CN106967630B (en) | Lactobacillus fermentum and application 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 |