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Lactobacillus helveticus strain and application thereof

A Lactobacillus and strain technology, applied in the field of microorganisms, can solve the problems of weak spleen and antidiarrheal function, hidden danger of drug-resistant gene proliferation, etc., to improve the structure and function of intestinal flora, no multidrug resistance, and safety. high effect

Active Publication Date: 2021-06-01
江苏斯卡露生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the technical problems in the prior art that probiotics can improve intestinal flora, invigorate the spleen and stop diarrhea, and have hidden dangers of drug-resistant gene diffusion, the present invention provides a strain of probiotics from natural pasture environment. After analyzing the morphological characteristics of the strain , physiological and biochemical characteristics, analysis and identification of molecular biological characteristics, the bacterial strain is Lactobacillus helveticus (Lactobacillus helveticus), it is named Lactobacillus helveticus-S

Method used

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  • Lactobacillus helveticus strain and application thereof
  • Lactobacillus helveticus strain and application thereof
  • Lactobacillus helveticus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Isolation, cultivation and identification of Lactobacillus helveticus-S

[0044] Screening of probiotics in soil collected from Yakeshi natural pasture in Hulunbuir, Inner Mongolia. Culture conditions: MRS medium, anaerobic culture at 37°C.

[0045] MRS medium is solid or liquid medium, and the formula of MRS liquid medium is: casein peptone 10.0g / L, beef extract 10.0g / L, yeast powder 5.0g / L, glucose 5.0g / L, sodium acetate 5.0g / L , diammonium citrate 2.0g / L, Tween 801.0g / L, K 2 HPO 4 2.0g / L, MgSO 4 ·7H 2 O 0.2g / L, MnSO 4 ·H 2 O 0.05g / L, CaCO 3 20.0g / L, pH6.8. MRS solid medium preparation method is to add agar 15.0g / L to MRS liquid medium.

[0046] The separation and screening method is as follows:

[0047] 1. Sampling: Use a sampling shovel to remove floating soil about 5cm from the surface, take 10-25g of soil samples at 5-25cm, and put them into plastic bags prepared in advance.

[0048] 2. Pour plate: The prepared MRS solid medium is sterilize...

Embodiment 2

[0066] Embodiment 2: gastric acid, bile salt tolerance of Lactobacillus helveticus-S

[0067] After the Lactobacillus helveticus-S strain was activated in the MRS liquid medium, it was transferred to the MRS liquid medium with a 5% inoculum size, and after 24 hours of anaerobic cultivation at 37°C, the thalline was collected by centrifugation at 6000rpm for 5 minutes, and washed with PBS (pH7.2 ) to wash the cells twice and finally resuspend the cells with PBS (pH7.2) and adjust the bacterial concentration to 1×10 8 CFU / mL. Take 50 ml of the above bacteria suspension and divide it into 5 parts on average, centrifuge at 6000rpm for 5 minutes to collect the bacteria, and use 10 ml of PBS (pH7.2), PBS (pH2.5), bile salt solution, artificial gastric juice, and artificial intestinal juice to weigh Suspended bacteria were anaerobically cultured at 37°C for 2 hours, plate counts were performed before and after culture, and the survival rate of bacteria was calculated. The results a...

Embodiment 3

[0076] Example 3: Hemolysis and drug resistance of Lactobacillus helveticus-S

[0077] 1. Hemolytic test: Lactobacillus helveticus-S was streaked and inoculated on a sheep blood plate, and cultured in a 37°C incubator for 24 hours to observe whether there was a hemolytic ring around the colony. No hemolytic ring appeared on the test blood plate, indicating that the Lactobacillus helveticus-S strain had no hemolytic property.

[0078] 2. Antibiotic susceptibility test: the concentration of Lactobacillus helveticus-S strain suspension was adjusted to 1×10 9 CFU / mL, take 1mL of the bacterial suspension and add it to a sterile petri dish, then add MRS solid medium that has been melted and cooled to about 45°C in the petri dish, mix well and place it horizontally. After solidification, the drug-sensitive paper was placed on the surface of the plate, and incubated in a 37°C incubator for 24 hours, and the diameter of the inhibition zone was measured with a vernier caliper. Drug su...

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Abstract

The invention provides a strain of probiotics from an environment, and through analysis and identification of morphological characteristics, physiological and biochemical characteristics and molecular biological characteristics of the strain, the strain is Lactobacillus helveticus and is named as Lactobacillus helveticus-S. The lactobacillus helveticus-S can resist acid and bile salt, so that the lactobacillus helveticus-S can be colonized in the digestive tract through oral administration; the lactobacillus helveticus-S has no hemolytic activity and high sensitivity to common antibiotics, so that the lactobacillus helveticus-S has no intestinal pathogenicity and does not diffuse antibiotic tolerance genes; animal model tests show that the lactobacillus helveticus-S can improve the structure and function of intestinal flora, improve the diversity of the intestinal flora and reduce the abundance of inflammation-related flora, and has the effects of treating diarrhea and loose stool and improving the symptom of spleen deficiency.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to an isolated probiotic strain and its use, in particular to a Lactobacillus helveticus-S strain isolated from a grassland pasture and its role in improving the structure and function of intestinal flora and treating spleen deficiency. use. Background technique [0002] Probiotics are normal flora in the intestinal tract, respiratory tract, reproductive tract, and mucous membrane system of humans and animals. Many research results have found that probiotics and intestinal mucosa together form a protective barrier to prevent the invasion of viruses and bacteria. Hentges et al. (1983) believed that the main function of the normal flora is to prevent the colonization of invading microorganisms in the intestinal tract. Gaskin et al. (1996) reported that normal flora can stimulate the development of animal mechanisms to enhance the body's immunity. Various antibacterial substances such...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P1/12A61P1/14A61P1/00A23L33/135C12R1/225
CPCA61K35/747A61P1/12A61P1/14A61P1/00A23L33/135A23V2002/00A23V2400/147A23V2200/32
Inventor 不公告发明人
Owner 江苏斯卡露生物科技有限公司
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