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Lactobacillus for producing lactic acid and H2O2 and application of lactobacillus

A technology of H2O2 and Lactobacillus, applied in the field of microorganisms, can solve the problems of low adhesion, weak ability, and inability to inhibit vaginal pathogenic bacteria well, and achieve the effect of inhibiting reproduction, good stability, and quickly restoring the vaginal microenvironment

Active Publication Date: 2020-05-01
SHANGHAI SINE PHARMA LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by this invention is in order to overcome Lactobacillus gasseri and Lactobacillus crispatus in the prior art to produce lactic acid and H 2 o 2 Defects such as weak ability, low adhesion ability and inability to well inhibit the reproduction of vaginal pathogenic bacteria provide a kind of lactic acid and H 2 o 2 Lactobacillus and its application

Method used

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  • Lactobacillus for producing lactic acid and H2O2 and application of lactobacillus
  • Lactobacillus for producing lactic acid and H2O2 and application of lactobacillus
  • Lactobacillus for producing lactic acid and H2O2 and application of lactobacillus

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Embodiment 1

[0036] The preparation of embodiment 1 culture medium

[0037] 1. Lactobacillus selective medium (Rogosa SL Agar) preparation:

[0038] (1) Weigh 22.5g of Rogosa SL Agar medium powder into a 500mL Erlenmeyer flask, add 300mL of deionized water, stir fully with magnetic force, then heat and boil for 1min under rapid stirring, and seal the bottle mouth;

[0039] (2) Put the above-mentioned sealed Erlenmeyer flask into a biological safety cabinet, and irradiate it with ultraviolet rays for more than 20 minutes;

[0040] (3) When the temperature of the medium drops to 50-60°C, open the bottle, add 396 μL of glacial acetic acid, put it in a microwave oven and boil for 2-3 minutes, and irradiate with ultraviolet light for more than 20 minutes;

[0041] (4) After the temperature of the culture medium has dropped to room temperature, pour it into a petri dish. Pour about 10-20mL of culture medium into each dish according to the size of the petri dish. After cooling and solidifying, m...

Embodiment 2

[0053] Example 2 Isolation, purification and enrichment culture of Lactobacillus gasseri and Lactobacillus crispatus strains

[0054] 1. Isolation, purification and enrichment culture of Lactobacillus strains

[0055] Several healthy women of childbearing age without vaginal infection or any intestinal disease were recruited to participate in providing samples. The participants all passed the health examination of the physical examination center, and provided information about their age (21-30), menstrual cycle and Information on other health behaviors, etc. Starting 2 weeks before the sample collection, all participants avoided all types of formulas containing probiotics. The samples were collected using the Port.A-Cd system of BD Company in the United States. Two sterile cotton swabs were used to collect the vaginal side wall of the subjects 1 / 3 of the secretion, put it into a sterile tube, and quickly transport it to the laboratory biosafety cabinet with an ice pack, rins...

Embodiment 3

[0065] Example 3 Determination of metabolites of Lactobacillus gasseri and Lactobacillus crispatus

[0066] 1. Determination of lactic acid content in the fermentation broth of Lactobacillus gasseri and Lactobacillus crispatus

[0067] D-lactic acid detection kit (Sigma) and L-lactic acid detection kit (Sigma) were used to measure the isolated 7 strains of Lactobacillus gasseri (or 7 strains of Lactobacillus crispatus) and the control strain Lactobacillus delbrueckii DM8909, rhamnose Lactobacillus PB01, Lactobacillus rhamnosus GR-1, ATCC standard strain 33820 (purchased from ATCC) were fermented in MRS medium for 48 hours and then the content of lactic acid in the fermentation broth. The measurement results show:

[0068] (1) Among the 7 strains of Lactobacillus gasseri, B3 had the strongest lactic acid-producing ability, followed by B7. -1) The lactic acid output is shown in Table 1.

[0069] (2) Among the 7 strains of Lactobacillus crispatus, A3 had the strongest lactic a...

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Abstract

The invention discloses lactobacillus for producing lactic acid and H2O2 and an application of the lactobacillus. The lactobacillus is lactobacillus gasseri or lactobacillus crispatus, wherein the preservation number of the lactobacillus gasseri is CGMCC No. 15815 or CGMCC No. 15816; the preservation number of the lactobacillus crispatus is CGMCC No. 15813 or CGMCC No. 15814. The lactobacillus gasseri and lactobacillus crispatus provided by the invention have extremely strong lactic acid and H2O2 production ability, have good ability to adhere to vaginal epithelial cells, and can effectively inhibit reproduction of pathogenic bacteria like Gardner bacteria and Candida albicans in in-vivo and in-vitro experiments.

Description

technical field [0001] The invention relates to the field of microbes, in particular to a method for producing lactic acid and H 2 o 2 Lactobacillus and its application. Background technique [0002] There are a variety of microorganisms in the vagina of healthy women, and they form a vaginal micro-ecological system that restricts, coordinates and dynamically balances with the host and the environment. It is reported in the literature that the vaginal flora of healthy women is mainly composed of Lactobacillus, and a certain type of Lactobacillus is absolutely dominant. According to the difference of the dominant Lactobacillus species in the vaginal flora, women can be divided into 5 categories: (1) Lactobacillus crispatus-dominant female, (2) Lactobacillus gasseri (also known as Lactobacillus gasseri)-dominant female, (3) Lactobacillus jansnii-dominant female, (4) Lactobacillus inertii dominant women, (5) women with diverse vaginal flora composition. Under normal circums...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04A61K35/747A61P31/04A61P31/10A61P33/02A61P15/02C12R1/225
CPCC12N1/20C12N1/04A61P31/04A61P31/10A61P33/02A61P15/02A61K2035/115C12R2001/225C12N1/205Y02A50/30
Inventor 徐晓芬付美红王莎莎高远尹培军朱刚刚王苒君于鸿晶孙宁云文彬
Owner SHANGHAI SINE PHARMA LAB
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