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A caries-resistant Lactobacillus plantarum and its application

A technology for Lactobacillus plantarum and caries, applied in the field of microorganisms, can solve problems such as staying, and achieve the effects of good survival ability, good inhibitory effect, good self-aggregation and copolymerization ability

Active Publication Date: 2020-08-28
SICHUAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on the above reports, some Lactobacillus plantarum has a potential control effect on dental caries, but at present it only stays in the in vitro test, and the control effect of lactic acid bacteria on caries in vitro does not directly indicate that it also has the same effect in vivo, so these strains There is still a certain distance to be applied to actual production

Method used

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  • A caries-resistant Lactobacillus plantarum and its application
  • A caries-resistant Lactobacillus plantarum and its application
  • A caries-resistant Lactobacillus plantarum and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Isolation, screening and molecular biology identification of Lactobacillus plantarum K41

[0032] 1. Material preparation

[0033] The samples were collected from homemade kimchi at a farmhouse on Yan'an Road, Shunqing District, Nanchong City, Sichuan Province;

[0034] The universal primer pair was synthesized by Invitrogen (Shanghai), and the 27F / 1492R sequence is as follows:

[0035] 27F: AGAGTTTGATCMTGGCTCAG

[0036] 1492R: TACGGYTACCTTGTTACGACTT

[0037] PBS buffer solution: Potassium dihydrogen phosphate 0.20g, disodium hydrogen phosphate 1.15g, sodium chloride 8.0g, potassium chloride 0.2g, add 800mL pure water and stir to dissolve, adjust concentrated hydrochloric acid to the corresponding pH value, and dilute to 1L.

[0038] 2. Isolation and screening of plant lactic acid bacteria with inhibitory effect on Streptococcus mutans and their molecular biological identification

[0039] Cut the collected sample into pieces, weigh 1 g, and put it into 9 ...

Embodiment 2

[0045] Example 2 Aggregation ability test of Lactobacillus plantarum K41

[0046] 1) Self-aggregation test: K41 and UA159 were cultured overnight at 37°C, centrifuged at 8000r / min for 10min, and the supernatant was discarded to collect the bacteria. Wash the bacteria twice with PBS buffer (same as Example 1) and resuspend, adjust the absorbance value OD of the bacteria suspension 600 0.6±0.02 (A 0 ). Take 2mL of the bacterial suspension and culture it at room temperature, and measure the OD of the supernatant every 1h 600 (At), a total of 4h was measured, with 3 repetitions in each group.

[0047] Self-aggregation ability = (A 0 -At) / A 0 ×100%

[0048] 2) Co-aggregation test: the preparation method of Streptococcus mutans and lactic acid bacteria suspension is the same as the self-aggregation ability test. Take 1 mL of lactic acid bacteria suspension (Ax) and 1 mL of Streptococcus 1h Measure the OD of the supernatant 600 (A(x+y)), measured for 4 hours in total, with 3 ...

Embodiment 3

[0053] Example 3 Lactobacillus plantarum K41 inhibits exopolysaccharide test

[0054] Adjust the suspension of UA159 and K41 bacteria activated for three generations to OD 600 =1±0.02, inoculate UA159 with 2% inoculation amount in BHI liquid medium containing 2% (w / v) sucrose, add an equal volume of K41 bacterial suspension and mix well, and use MRS medium without bacterial liquid as blank For the control, cultured at 37°C for 24 hours, and each group had 3 replicates. Anthrone-sulfuric acid method was used to measure the content of exopolysaccharide, glucose was used as a standard curve, and then the content of exopolysaccharide was calculated from the standard curve.

[0055] The test results showed that: Lactobacillus plantarum K41 could significantly inhibit the formation of Streptococcus mutans exopolysaccharide, the inhibition rate of soluble exopolysaccharide was 44.06%, and the inhibition rate of insoluble exopolysaccharide was 90.70%.

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Abstract

The invention discloses a caries-resistant plant lactobacillus ( Lactobacillus plantarum ) K41 and its applications. The Lactobacillus plantarum K41 provided by the present invention was preserved in the General Microorganism Center of the China Microbiological Culture Collection Management Committee, with a preservation number of CGMCC No. 15462 and a preservation date of March 20, 2018. The strain was isolated from farmhouse-made kimchi in Yan'an Road, Shunqing District, Nanchong City, Sichuan Province. It grew well on MRS agar medium and could inhibit the growth of Streptococcus mutans UA159 cells, exopolysaccharide formation, hydroxyapatite degradation, and biofilm It has good self-polymerization and copolymerization ability, good tolerance to antibiotics, salt and acid, and is easy to colonize in the oral cavity. Animal experiments have shown that K41 can inhibit the demineralization of rat tooth enamel and the degree of dental caries, and its application in functional foods is of great significance to human health.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a plant lactic acid bacterium capable of controlling dental caries and its application in the preparation of functional food. Background technique [0002] Dental caries, also known as "tooth decay" or "tooth decay", is a common bacterial infectious disease in the human oral cavity. The World Health Organization has listed it as one of the three major human diseases to prevent and control, along with tumors and cardiovascular diseases. According to the Global Burden of Disease Study 2016 (GBD) data published by the world's authoritative medical journal "The Lancet", the global prevalence of permanent dental caries ranks first among all diseases, and the incidence rate ranks second , second only to upper respiratory tract infection, the incidence of deciduous dental caries ranks fifth. According to the results of the 4th national oral health epidemiological survey released...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L33/135C12R1/25
CPCA23L33/135A23V2002/00C12R2001/25C12N1/205A23V2400/169A23V2200/312
Inventor 孙群张国建李雨庆卢淼刘绒梅李洋
Owner SICHUAN UNIV
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