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Lactobacillus salivarius LS97 and application thereof

A technology of Lactobacillus salivarius and strains, which is applied in the field of microbiology, can solve the problems of blank strain resources and achieve excellent results and alleviate dental caries

Active Publication Date: 2020-02-21
JIANGSU WECARE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it should be pointed out that there are many reports on the breeding of strains that can antagonize the oral caries bacteria Streptococcus mutans, but there are almost no reports on the screening of strains that can efficiently inhibit the cousin Streptococcus, and the resources of related strains are almost blank

Method used

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  • Lactobacillus salivarius LS97 and application thereof
  • Lactobacillus salivarius LS97 and application thereof
  • Lactobacillus salivarius LS97 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The breeding of embodiment 1 Lactobacillus salivarius LS97 obtains

[0022] 1) Strain screening

[0023] Extract 5mL of the diluent of healthy infant digestive tract advance solution from Bama Town, Hechi City, Guangxi Zhuang Autonomous Region, China (the crowd had never taken antibiotics, had no history of probiotics, and had no history of gastrointestinal diseases before collection), and was serially diluted 10 times to 10 -6 , 200 μL of each dilution concentration was spread on the selective MRS medium, anaerobically cultured at 37°C for 48-72h, and different single colonies on the plate were isolated and obtained. The above-mentioned different single colonies were picked with a sterilized inoculation loop, streaked and purified on the MRS solid medium, and cultured anaerobically at 37°C for 48 hours to obtain a total of 11 pure lactic acid bacteria strains, which were named A1-A11.

[0024] 2) The test of isolated bacterial strains against cousin Streptococcus

...

Embodiment 2

[0042] Embodiment two strain identification:

[0043] According to the bacterial strain that embodiment one obtains, identification method is as follows:

[0044] The obtained strain A5 was identified by 16s rDNA sequencing and physiological and biochemical identification. The 16s rDNA universal primer sequence 27F was: 5'-AGAGTTTGATCCTGGCTCAG-3' (Seq ID No: 1), and 1492R was: 5'-GGTTACCTTGTTACGACTT-3' (Seq ID No: 2). The 16S rDNA gene sequence of strain A5 was amplified and sequenced, and the PCR amplification product was sent to Shanghai Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing. The nucleotide sequence of 16S rDNA of strain A5 is shown in Seq ID No: 3 . After 16s rDNA gene comparison, the similarity rate with the Lactobacillus salivarius strain in Genebank was 100%. Combined with the physiological and biochemical identification results, strain A5 was identified as a strain of Lactobacillus salivarius, named Lactobacillus salivarius LS97, and its physiologi...

Embodiment 3

[0048] Embodiment Lactobacillus trisalivarius LS97 is to the inhibitory ability analysis of Streptococcus mutans and Streptococcus sanguis growth

[0049] Similarly, the double-layer cup and saucer method was used to investigate the inhibition of Lactobacillus salivarius LS97 on the growth of Streptococcus mutans UA159 (purchased from ATCC) and Streptococcus sanguinis (Streptococcus sanguinis SK36, purchased from ATCC). Pour the agar medium into the plate, fix the aseptic Oxford cup in a suitable position after solidification, then mix 0.7% (w / v) BHI medium with the mixture containing Streptococcus mutans or Streptococcus sanguis (10 7 CFU / mL) was poured on the lower medium. After coagulation, take out the Oxford cup, and add 100 μL of Lactobacillus culture supernatant to each well. Incubate the plate above at 37°C (5% CO 2 ) overnight, and a previously reported strain of Streptococcus salivarius with the ability to inhibit the above two strains was used as a control strain ...

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Abstract

The invention relates to the field of microorganisms, and in particular to lactobacillus salivarius LS97 and an application thereof. The strain is preserved in the China General Microbiological Culture Collection Center on Dec.10, 2018, and the preservation number of the strain is CGMCC NO.16922. The strain is classified and named as Lactobacillus salivarius, and the preservation address of the strain is Institute of Microbiology, Chinese Academy of Sciences on No.3, No.1 Courtyard, Beichen West Road, Chaoyang District, Beijing. The lactobacillus salivarius LS97 can effectively inhibit growthof streptococcus sobrinus.

Description

technical field [0001] The invention relates to the field of microbes, in particular to a Lactobacillus salivarius LS97 and an application thereof. Background technique [0002] Cousin Streptococcus (Streptococcus sobrinus) used to be known as distant Streptococcus or Streptococcus pilosa, so named because it has a hairy layer under the electron microscope. For a long time, cousin Streptococcus has been considered as an important cariogenic bacteria. Compared with Streptococcus mutans, Cousin Streptococcus has stronger acid production and acid resistance, and is more closely related to high caries. Therefore, it has been paid more and more attention by researchers. [0003] At present, the methods to prevent and treat dental caries caused by cousin streptococcus mainly include drug method, diet control method and mechanical removal method, etc., for example, the use of compounds containing fluoride, phenolic compounds or strong oxidants can be used to kill the cousin strept...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P1/02C12R1/225
CPCA61K35/747A61P1/02C12R2001/225C12N1/205
Inventor 方曙光吴明科朱建国王思清占英英
Owner JIANGSU WECARE BIOTECHNOLOGY CO LTD
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