CN112391305A

CN112391305A - Streptococcus salivarius F286 and uses thereof

Info

Publication number: CN112391305A
Application number: CN201910754930.9A
Authority: CN
Inventors: 申剑; 李娜
Original assignee: Shanghai Jiaotong University
Current assignee: Shanghai Jiaotong University
Priority date: 2019-08-15
Filing date: 2019-08-15
Publication date: 2021-02-23
Anticipated expiration: 2039-08-15
Also published as: CN112391305B

Abstract

The invention provides streptococcus salivarius F286 with a preservation number of CGMCC No. 17579. In the invention, streptococcus salivarius F286 is obtained by the inventor through human breast milk and is perfused into 8-week-old sterile mice growing in a sterile bag, and the same breast milk is used for carrying out secondary perfusion on each mouse every other day. After the completion of the gavage, the mice were further kept in a sterile bag for 8 weeks, and the mice were subjected to bacterial isolation using the feces of the mice at the 8 th week. The streptococcus salivarius F286 can be used as probiotic preparation, food additive, or effective component of medicine, especially for infantile breast milk, breast milk substitute, infant formula, dairy product or related product lacking streptococcus.

Description

Streptococcus salivarius F286 and uses thereof

Technical Field

The invention relates to a bioengineering bacterium, in particular to streptococcus salivarius F286 and application thereof.

Background

Human breast milk not only provides nutrition for the newborn but also human symbiotic bacteria, and studies have shown that milk of healthy mothers contains 10²–10⁵CFU/ml bacteria. According to each pure breast-fed infantThe average daily intake is calculated by 800 ml of breast milk, which is provided to them 10 days⁴–10⁸And (4) bacterial cells. Bacteria in breast milk colonize the newborn intestinal tract immediately after birth, are the "pioneer flora" of the newborn intestinal tract, and play a key role in the development of the newborn immune system.

Existing research has focused mainly on isolating bifidobacteria (bifidobacteria) and lactobacilli (lactobacilli) from breast milk and investigating the immunomodulatory effects of these traditional probiotics. However, not all breast milk contains viable bifidobacteria and lactobacilli which can be isolated in culture, and studies of the flora structure of breast milk in larger, larger populations have shown that the bacterium which is most abundant in breast milk and which is detectable in breast milk of different people is Streptococcus (Streptococcus). Jost et al collected fresh milk from 7 mothers, only bifidobacteria were isolated from 2 of them, lactobacilli from 1 mother, but streptococci from all 7 mothers; moreover, the literature indicates that, among the bacteria isolated from human breast milk, bifidobacteria account for only 1.7% and streptococci for 17.9%. In addition, streptococcus is also the predominant bacterium in the gut of newborns within 2 weeks after birth. These results suggest that breast milk streptococcus is a human commensal bacterium that most mothers transmit to their offspring through milk and has important probiotic effects. Therefore, there is a need to provide probiotic formulations of streptococcus in human breast milk.

Disclosure of Invention

In view of the above-mentioned drawbacks of the prior art, it is an object of the present invention to provide streptococcus salivarius F286 for solving the problem of streptococcus deficiency in some breast milk, breast milk substitute, infant formula, dairy products or related products in the prior art, and its use.

In order to achieve the above objects and other related objects, the invention provides streptococcus salivarius F286 with a preservation number of CGMCC No. 17579.

In the invention, streptococcus salivarius F286 is obtained by the inventor through human breast milk and is perfused into 8-week-old sterile mice growing in a sterile bag, and the same breast milk is used for carrying out secondary perfusion on each mouse every other day. After the completion of the gavage, the mice were further kept in a sterile bag for 8 weeks, and the mice were subjected to bacterial isolation using the feces of the mice at the 8 th week.

The strain is determined to be streptococcus salivarius through 16S rRNA gene amplification sequence development tree analysis, and according to the international naming rule: the strain is named by the genus name, the species name and the strain name, the genus name, the species name and the strain name are respectively streptococcus, streptococcus salivarius strains and F286, the strain is named as streptococcus salivarius F286, the strain is sent to a common microorganism center of China Committee for microorganism preservation management in 2019 and 19 months, and the preservation number is as follows: CGMCC No. 17579.

Further, the nucleotide sequence of the streptococcus salivarius F286 is shown as SEQ ID NO. 5.

The invention also provides application of the streptococcus salivarius F286 in preparing food, health-care products or medicines.

Further, the food or health product may be specific for infants, and may be, for example, a probiotic preparation, an infant formula milk powder, or a liquid beverage, such as a dairy product.

Further, the streptococcus salivarius F286 can be live or killed.

In another aspect of the invention, a food or health product is provided, which comprises streptococcus salivarius F286 or a streptococcus salivarius F286 lysate.

Further, the food or health product may be specific for infants, for example, it may be a probiotic preparation, an infant formula, or a liquid beverage, such as a dairy product.

Further, the streptococcus salivarius F286 can be live or killed.

Further, the content of streptococcus salivarius F286 in the food or health product is 10²CFU/ml-10¹⁰CFU/ml。

Further, the food or health care product has the functions of promoting the development of mammals, activating the immune function of the mammals and/or improving the immunity of the mammals.

Further, the food or health care product has the function of promoting peripheral blood mononuclear cells of mammals to secrete Interleukin (IL) -12 and Interleukin (IL) -10.

According to a further aspect of the invention there is provided a medicament comprising streptococcus salivarius F286 or a streptococcus salivarius F286 lysate as hereinbefore described.

Further, the streptococcus salivarius F286 can be live or killed.

Further, the medicine has the function of promoting the secretion of Interleukin (IL) -12 and Interleukin (IL) -10 by peripheral blood mononuclear cells of mammals.

Further, the medicament has the effects of promoting the development of mammals, activating the immune functions of the mammals and/or improving the immunity of the mammals.

Further, the medicament may be for use with infants and young children.

As described above, streptococcus salivarius F286 and uses thereof according to the present invention have the following beneficial effects:

the streptococcus salivarius F286 can be used as a probiotic preparation, a food additive or an effective component of a medicament, and particularly aims at the condition that streptococcus is lacked in infant breast milk, breast milk substitutes, infant formula milk, dairy products or related products. Streptococcus salivarius F286 is safe and effective. Experiments prove that the compound can effectively promote human peripheral blood mononuclear cells to secrete Interleukin (IL) -12 and Interleukin (IL) -10, and can also promote the expression of immune genes of nematodes and prolong the life of the nematodes.

The preservation information of the strain of the invention is as follows:

the strain name: streptococcus salivarius

The preservation number is as follows: CGMCC No. 17579;

the preservation date is as follows: 19/4/2019;

the name of the depository: china general microbiological culture Collection center;

the preservation unit is abbreviated as: CGMCC;

the address of the depository: xilu No.1 Hospital No.3, Beijing, Chaoyang, North.

Drawings

FIG. 1 shows the F286ERIC map of Streptococcus salivarius.

FIG. 2 shows the evolutionary tree of S.salivarius F286, based on the full-length sequence of the 16S rRNA gene.

FIG. 3 shows that the amount of Interleukin (IL) -12 and Interleukin (IL) -10 secreted by the F286 thalli cells of Streptococcus salivarius is stimulated by human Peripheral Blood Mononuclear Cells (PBMC), and the stimulation of the F286 thalli cells of Streptococcus salivarius is stronger than that of the existing commercial probiotic Lactobacillus rhamnosus GG (LGG).

FIG. 4 shows that the somatic cells of Streptococcus salivarius F286 significantly prolong the life of C.elegans (Caenorhabditis elegans), and the capacity of the somatic cells of Streptococcus salivarius F286 to prolong the life of nematodes is comparable to that of the existing commercial probiotic Lactobacillus rhamnosus GG (LGG).

FIG. 5 shows that the streptococcus salivarius F286 bacterial cells significantly improve the expression level of nematode (Caenorhabditis elegans) immunity genes, and the capability of the streptococcus salivarius F286 bacterial cells in improving the expression of the nematode immunity genes is equivalent to that of the existing commercial probiotic lactobacillus rhamnosus GG (LGG).

Detailed Description

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.

Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.

Unless otherwise indicated, the experimental methods, detection methods, and preparation methods disclosed herein all employ techniques conventional in the art of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related arts. These techniques are well described in the literature, and may be found in particular in the study of the MOLECULAR CLONING, Sambrook et al: a LABORATORY MANUAL, Second edition, Cold Spring Harbor LABORATORY Press, 1989and Third edition, 2001; ausubel et al, Current PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, 1987and periodic updates; the series METHODS IN ENZYMOLOGY, Academic Press, San Diego; wolffe, CHROMATIN STRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; (iii) METHODS IN ENZYMOLOGY, Vol.304, Chromatin (P.M.Wassarman and A.P.Wolffe, eds.), Academic Press, San Diego, 1999; and METHODS IN MOLECULAR BIOLOGY, Vol.119, chromatography Protocols (P.B.Becker, ed.) Humana Press, Totowa, 1999, etc.

Materials and sources thereof:

sterile mice: shanghai Slek's laboratory animal center, C57BL/6J mice

Wilkins-Chalgren (WCH) medium: qingdao, Haibop; the goods number is: HB0261

M17 medium: qingdao, Haibop; the goods number is: HB0391

The NGM culture medium is prepared by the following reagents:

NaCl,Sigma S7653-1KG

Peptone，Sigma P6713-500G

Agar，Sigma 05038-500G

CaCl₂.2H₂O,Sigma C3881-500G

MgSO₄.7H₂O，Sigma M1880-500G

KH₂PO₄,Sigma P0662-500G

K₂HPO₄.3H₂O，Sigma,P9666-500G

Cholesterol，Sigma C8667-1G

the NGM culture medium configuration method comprises the following steps: weighing 3g of NaCl, 2.5g of Peptone and 20g of Agar, adding 975mL of single distilled water for dissolution, sterilizing by high-pressure steam at 121 ℃ for 20min, and cooling to about 65 ℃. Under sterile conditions, the following sterile solutions were added: 1mL of 1M CaCl₂，1mL 1M MgSO₄25mL of 1M potassium phosphate buffer (pH 6.0), 1mL of 5mg/mL Cholesterol. Mixing, subpackaging with sterile pipette into 6 cm culture dish (15 mL each), standing at room temperature for solidification to obtain NGM agar plate

Nematodes: american nematode Collection CGC (Camphorhabditis Genetics Centre, University of Minnesota, Minneapolis)

Escherichia coli OP 50: american nematode Collection CGC (Camphorhabditis Genetics Centre, University of Minnesota, Minneapolis)

Example 1 isolation of Streptococcus salivarius F286

100 microliters of freshly collected human breast milk was gavaged into 8-week-old sterile mice grown in sterile bags, and each mouse was gavaged twice every other day with the same breast milk. Mice were kept on sterile bags for 8 weeks after the gavage was completed. Bacterial isolation was performed with mouse feces from week 8.

1mL of sterile 0.01mol/L phosphate buffer (PBS, 0.1% L-Cysteine was added) was immediately added to the mouse feces, and the mixture was sufficiently shaken to make the feces sample uniformly turbid. Making the fecal suspension into 10^-2，10^-3，10^-4，10^-5，10^-6， 10^-7，10^-8，10^-9Dilution gradients, 100. mu.L each of the dilutions of each gradient were plated on two solid medium plates (1.2% agar) of Wilkins-Chalgren (WCH) and M17, respectively, and three plates were repeated for each dilution gradient. All plates were placed upside down in an anaerobic incubator and incubated at 37 ℃. After 48 hours of culture, the plate with moderate colony number and more separable single colonies is selected. Colonies were randomly picked on each plate of each medium according to their colony morphology and size, transferred to the corresponding new WCH and M17 solid medium plates, respectively, and numbered. Each single colony was streaked three times for purification. The purified single colony is picked into a liquid M17 culture medium, enrichment culture is carried out for 24 hours, and the obtained culture is preserved and subsequently identified.

Example 2 identification of Streptococcus salivarius F286

(1) Extracting genome DNA: after 24 hours of anaerobic liquid culture, 3mL of the bacterial solution was collected, 9000g of the solution was centrifuged for 5 minutes, 475. mu.L of TE buffer (10mM Tris-HCl,1mM EDTA, pH 8.0) and 25. mu.L of lysozyme (lysozyme, 50mg/mL) were added, shaking and incubation were carried out at 37 ℃ for 1 hour, 5. mu.L (20. mu.g/mL) of proteinase K and 50. mu.L (20%) of SDS were added, shaking and mixing were carried out, and then incubation was carried out at 55 ℃ for 30 minutes. Adding phenol chloroform isoamyl alcohol (volume ratio 25:24:1) with the same volume of about 550 mu L, shaking and mixing uniformly, standing at 4 ℃, centrifuging at 14000rpm for 15min, taking supernatant, and repeatedly extracting with chloroform isoamyl alcohol (volume ratio 24:1) for 1-2 times. The supernatant was added with two volumes (about 800. mu.L) of absolute ethanol previously placed at-20 ℃ and 80. mu.L (3M) of sodium acetate, and left to stand in a refrigerator at-20 ℃ for 2 hours to precipitate DNA, and centrifuged at 14000rpm for 15min to collect DNA. After drying at low temperature in vacuo, the mixture was dissolved in 50. mu.l of TE buffer (Tris-HCl, pH8, 10 mM). Mu.l RNase (20mg/mL) was added thereto, and the mixture was gently mixed and incubated at 37 ℃ for 30min to digest RNA. DNA concentration was quantified using a microplate reader SpectraMax M5(Molecular Devices, San Francisco, USA) in combination with PicoGreen fluorescent dye (Thermo Fisher Scientific, Sunnyvale, USA).

(2) ERIC-PCR (Enterobacterial reliable interactive consensus sequence-PCR) fingerprinting: eThe upstream of the amplification primer of RIC-PCR is ERIC1(SEQ ID NO. 1: 5'-atgtaagctcctggggattcac-3'), the downstream is ERIC2((SEQ ID NO. 2: 5'-aagtaagtgactggggtgagcg-3'). 25 muL of PCR amplification system contains 20ng of bacterial genome DNA, the concentrations of four deoxynucleotides (dNTPs) are respectively 200mM,2.5U of TaKaRa rTaq DNA polymerase (Takara, Dalian, China),1 XPCR buffer (Mg2+ free),2mM MgCl₂Primers were 10pM each. The PCR procedure was: pre-denaturation at 95 ℃ for 7 min; denaturation at 95 deg.C for 30s, annealing at 52 deg.C for 1min, extension at 65 deg.C for 8min, and circulation for 30 times; finally, extension is carried out for 16min at 65 ℃. 400ng of ERIC-PCR product was subjected to 1.5% (w/v) agarose electrophoresis using a UVI gel imaging system (Tanon 3500, Tanon science)&Technology co., ltd., China) was photographed to obtain a fingerprint (fig. 1).

The ERIC map is the embodiment of the specific genome of the strain and can be used as the characteristic map of the streptococcus salivarius F286 strain.

(3) PCR amplification, cloning, sequencing and evolutionary position analysis of 16S rRNA gene full-length sequence of streptococcus salivarius F286CGMCC No.17579 strain:

primers used for PCR amplification of the full-length sequence of the 16S rRNA gene of the strain were 27f (SEQ ID NO. 3: 5'-agagtttgatcctggctcag-3') and 1492r (SEQ ID NO. 4: 5'-cggcttaccttgttacgactt-3'). A25. mu.L system included 0.75U rTaq DNA polymerase (Takara, Dalian, China),1 XPCR buffer (Mg2+ free),2mM MgCl₂10pmol of each primer, 200. mu.M of each of the four deoxynucleotides, and 10ng of bacterial genomic DNA as a template. The amplification procedure was as follows: pre-denaturation at 95 ℃ for 7 min; denaturation at 94 deg.C for 30s, annealing at 52 deg.C for 1min, and extension at 65 deg.C for 8min, and repeating the above steps for 25 times; finally, extension was carried out at 65 ℃ for 16 min.

The PCR product was purified according to the instructions of the Gel Extraction Kit 200(Omega, USA). The purified PCR product was ligated with the Vector pGEM-T Easy Vector (Promega, Madison, USA) according to the ligation kit instructions, and the ligation product was transformed into competent cells of the host bacterium E.coli DH5 alpha (Transgen, Beijing, China). Then, the cells were spread on LB-ampicillin (100. mu.g/mL) plates to which IPTG and X-Gal were added in advance at a predetermined concentration, cultured at 37 ℃ for 12 hours, and white spot positive clones were randomly selected and sequenced (Life Technologies, Shanghai, China). The full-length sequence of the 16S rRNA gene of the streptococcus salivarius F286 strain is shown in SEQ ID NO. 5:

BLAST (http:// www.ncbi.nlm.ni h.gov/BLAST) alignment of the obtained 16S rRNA gene sequences in the Genbank database was performed, and the bacteria having the most similar sequences to the known bacteria in the database were: streptococcus salivaria strain NCTC8618 has a similarity of 99.93%. A phylogenetic tree (Neighbor-joiningphylogenetic tree) was constructed using MEGA 5 software (Molecular evolution Genetics Analysis package), showing that the F286 strain is S.salivarius (FIG. 2).

Example 3 Heat-killed bacterial cells of Streptococcus salivarius F286 promote secretion of Interleukin 12(IL-12) and Interleukin 10(IL-10) by human Peripheral Blood Mononuclear Cells (PBMCs)

Streptococcus salivarius F286 and lactobacillus rhamnosus LGG were cultured in M17 (china, Qingdao, Haibo) and MRS (china, Qingdao, Haibo) liquid media, respectively, for 8 hours, to reach the plateau phase. The culture was centrifuged at 5,000g for 10min to collect the cells of Streptococcus salivarius F286 and Lactobacillus rhamnosus LGG. After resuspending the somatic cells with PBS, 5,000g of the cells were centrifuged for 10min, and the cells were washed 2 times to remove the bacterial culture medium. The cell concentration of the cells was adjusted to 10 with PBS⁸CFU/ml and 10⁹CFU/ml, water bath at 65 deg.C for 20min, killing thallus cells by heat, and freezing to-80 deg.C for use.

In 24-well plates, 2X 10 inoculations per well⁶After the addition of 20. mu.l of 1X 10 cells (PBMC) to individual Peripheral Blood Mononuclear Cells (PBMC)⁸CFU/ml or 1X 10⁹The final volume of the CFU/ml heat-killed streptococcus salivarius F286 thallus and the Lactobacillus rhamnosus LGG thallus is 1ml, and the ratio of cells to bacteria is respectively 1:1 and 1: 10 in a co-incubation system. In the negative control group, no bacterial cells were added, and 20. mu.l of PBS was added. The cell culture medium contains 10% fetal calf serum and 1% streptomycin and ampicillin1640 cell culture medium of penicillin. The mixture was incubated at 37 ℃ in a 5% carbon dioxide incubator for 24 hours. The cell culture supernatants were collected and tested for interleukin 10 and interleukin 12 concentrations using an ELISA kit.

As shown in FIG. 3, the bacterial cells of Streptococcus salivarius F286 stimulated more Interleukin (IL) -12 and Interleukin (IL) -10 secretion from human Peripheral Blood Mononuclear Cells (PBMC) (FIG. 3), while the bacterial cells of Lactobacillus rhamnosus LGG stimulated only a small amount of Interleukin (IL) -12 and Interleukin (IL) -10 secretion from human Peripheral Blood Mononuclear Cells (PBMC).

Example 4 somatic cells of Streptococcus salivarius F286 increase expression levels of nematode immunity genes and prolong nematode longevity

(1) Somatic cells of streptococcus salivarius F286 prolong life of nematodes

Streptococcus salivarius F286 and Lactobacillus rhamnosus LGG were cultured in M17 medium (China, Qingdao, Haibo) and MRS medium (China, Qingdao, Haibo) respectively for 8 hours at 37 ℃ anaerobic workstation (DG500, DWS, United Kingdom). Escherichia coli OP50 was cultured overnight at 37 ℃ in an aerobic condition. Centrifuging the cultured bacterial liquid for 10min at 15000 Xg, removing supernatant, collecting thallus of streptococcus salivarius F286, lactobacillus rhamnosus LGG and escherichia coli OP50, centrifuging for 10min under the condition of 15000 Xg by using sterile M9 buffer solution, and washing the thallus twice. The washed cells were adjusted to a cell concentration of 10 mg/100. mu.l (wet weight/volume) with M9 buffer solution and mixed well. 100. mu.l of bacterial resuspension containing 10mg of cells was added dropwise to a 6 cm diameter NGM (mNGM) plate without peptone addition.

After the nematodes growing to L3 on peptone-supplemented NGM plates and feeding on E.coli OP50, they were transferred to mNGM plates, feeding on Streptococcus salivarius F286, Lactobacillus rhamnosus LGG and E.coli OP50 respectively, and incubated at 25 ℃. Streptococcus salivarius F286, Lactobacillus rhamnosus LGG and Escherichia coli OP50 were individually plated on 5 parallel plates (20-25 plates) for a total of 100-125 nematode species for life testing and comparison. During the experiment, the nematode was considered dead in the absence of a mild stimulation of the platinum wire. Kaplan-Meier Survival Analysis was performed using OASIS 2(Online Application for surveyal Analysis 2) on-line software, and log-rank was used to test the difference in mean life span of nematodes fed to different bacterial strains.

The results are shown in fig. 4, where both somatic cells of streptococcus salivarius F286 and LGG cells of lactobacillus rhamnosus significantly prolonged the life span of the nematodes, compared to the standard food e.coli OP50 for nematodes; meanwhile, the two have no difference in the ability of prolonging the life of the nematode. The safety and the probiotics of the streptococcus salivarius F286 are illustrated.

(2) Extraction of RNA of nematode of thallus cell of streptococcus salivarius F286

After feeding Streptococcus salivarius F286, Lactobacillus rhamnosus LGG, or Escherichia coli OP50 on day 14, 500 nematodes were collected, the nematodes were lysed by TRIZOL reagent (Invitrogen) to extract RNA, the RNA was purified by RNeasy Mini Kit (Qiagen), and residual DNA was removed by DNaseI (Invitrogen) Kit.

(3) qPCR method for determining expression level of nematode immunity gene improved by somatic cells of streptococcus salivarius F286

The purified nematode RNA was used in SuperScriptTMfirst-Strand Synthesis System for RT-PCR (Invitrogen) reverse transcription kit, and oligo (dT) was used as a primer to synthesize the first cDNA Strand. Quantitative detection of nematode immunity genes was performed using SYBR Green Supermix (BIO-RAD) (Roche, LightCycler 96). Using housekeeping gene act-1 as reference gene and 2^-ΔΔCtThe relative expression level of the immune gene was calculated (see FIG. 5).

The results show that compared with the standard food Escherichia coli OP50 of nematodes, the somatic cells of Streptococcus salivarius F286 significantly improve the expression level of nematode immunity genes (cpr-1, cpr-5, lys-5, lys-7, clec-60, clec-85, C15C8.3), and the capacity of the somatic cells of Streptococcus salivarius F286 for improving the expression of the nematode immunity genes is equivalent to that of Lactobacillus rhamnosus LGG.

The above examples are intended to illustrate the disclosed embodiments of the invention and are not to be construed as limiting the invention. In addition, various modifications of the methods and compositions set forth herein, as well as variations of the methods and compositions of the present invention, will be apparent to those skilled in the art without departing from the scope and spirit of the invention. While the invention has been specifically described in connection with various specific preferred embodiments thereof, it should be understood that the invention should not be unduly limited to such specific embodiments. Indeed, various modifications of the above-described embodiments which are obvious to those skilled in the art to which the invention pertains are intended to be covered by the scope of the present invention.

Sequence listing

<110> Shanghai university of transportation

<120> Streptococcus salivarius F286 and uses thereof

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1470

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

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gacgaacgct ggcggcgtgc ctaatacatg caagtagatc gctgaagaga ggagcttgct 60

cttcttggat gagttgcgaa cgggtgagta acgcgtaggt aacctgcctt gtagcggggg 120

ataactattg gaaacgatag ctaataccgc ataacaatgg atgacacatg tcatttattt 180

gaaaggggca attgctccac tacaagatgg acctgcgttg tattagctag taggtgaggt 240

aacggctcac ctaggcgacg atacatagcc gacctgagag ggtgatcggc cacactggga 300

ctgagacacg gcccagactc ctacgggagg cagcagtagg gaatcttcgg caatgggggc 360

aaccctgacc gagcaacgcc gcgtgagtga agaaggtttt cggatcgtaa agctctgttg 420

taagtcaaga acgagtgtga gagtggaaag ttcacactgt gacggtagct taccagaaag 480

ggacggctaa ctacgtgcca gcagccgcgg taatacgtag gtcccgagcg ttgtccggat 540

ttattgggcg taaagcgagc gcaggcggtt tgataagtct gaagttaaag gctgtggctc 600

aaccatagtt cgctttggaa actgtcaaac ttgagtgcag aaggggagag tggaattcca 660

tgtgtagcgg tgaaatgcgt agatatatgg aggaacaccg gtggcgaaag cggctctctg 720

gtctgtaact gacgctgagg ctcgaaagcg tggggagcga acaggattag ataccctggt 780

agtccacgcc gtaaacgatg agtgctaggt gttggatcct ttccgggatt cagtgccgca 840

gctaacgcat taagcactcc gcctggggag tacgaccgca aggttgaaac tcaaaggaat 900

tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc 960

ttaccaggtc ttgacatccc gatgctattt ctagagatag aaagttactt cggtacatcg 1020

gtgacaggtg gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc 1080

aacgagcgca acccctattg ttagttgcca tcattcagtt gggcactcta gcgagactgc 1140

cggtaataaa ccggaggaag gtggggatga cgtcaaatca tcatgcccct tatgacctgg 1200

gctacacacg tgctacaatg gttggtacaa cgagttgcga gtcggtgacg gcaagctaat 1260

ctcttaaagc caatctcagt tcggattgta ggctgcaact cgcctacatg aagtcggaat 1320

cgctagtaat cgcggatcag cacgccgcgg tgaatacgtt cccgggcctt gtacacaccg 1380

cccgtcacac cacgagagtt tgtaacaccc gaagtcggtg aggtaacctt ttggagccag 1440

ccgcctaagg tgggatagat gattggggtg 1470

Claims

1. A streptococcus salivarius F286, wherein: the preservation number is CGMCC No. 17579.

2. Streptococcus salivarius F286 as claimed in claim 1 wherein: the nucleotide sequence of the streptococcus salivarius F286 is shown in SEQ ID NO. 5.

3. Use of streptococcus salivarius F286 as claimed in any of claims 1 to 2 in the manufacture of a food, health product or pharmaceutical product.

4. Use according to claim 3, characterized in that: the streptococcus salivarius F286 is live or killed.

5. A food, health care product or pharmaceutical product, wherein the food, health care product or pharmaceutical product comprises streptococcus salivarius F286 or a streptococcus salivarius F286 lysate.

6. The food, health product or pharmaceutical product according to claim 5, wherein: the streptococcus salivarius F286 is live or killed.

7. The food, health product or pharmaceutical product according to claim 5, wherein: the content of streptococcus salivarius F286 in the food, health product or medicine is 10²CFU/ml-10¹⁰CFU/ml。

8. The food, health product or pharmaceutical product according to claim 5, wherein: the food, health product or medicine has functions of promoting mammal peripheral blood mononuclear cell to secrete Interleukin (IL) -12 and Interleukin (IL) -10.

9. The food, health product or pharmaceutical product according to claim 5, wherein: the food, health product or medicine is suitable for infant.

10. The food, health product or pharmaceutical product according to claim 5, wherein: the food or health product is a probiotic preparation, infant formula milk powder or liquid beverage.