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A kind of lactobacillus plantarum and application thereof

A technology of Lactobacillus plantarum and strains, which is applied in the field of microorganisms and can solve problems such as weak inhibitory ability

Active Publication Date: 2021-10-01
天益健康科学研究院(镇江)有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Lactobacillus plantarum 229v and Lactobacillus plantarum Lp-115 are two Lactobacillus plantarum strains certified by the US Food and Drug Administration (FDA) as generally recognized as safe (GRAS), Lactobacillus plantarum 229v and Lactobacillus plantarum Lp-115 has a certain ability of acid resistance and bile salt resistance, so it is widely used. However, the applicant finds that it is different in acid resistance, bile salt resistance, and the ability to inhibit the growth of pathogenic bacteria. For example, Lactobacillus plantarum NKK20 has different The adhesion rate of intestinal mucosa is significantly higher than that of Lactobacillus plantarum 229v; for another example, Lactobacillus plantarum 229v has weak inhibitory ability to Staphylococcus aureus ATCC25923; Lactobacillus plantarum LP-115 has weak inhibitory ability to Salmonella enteritidis ATCC 14028

Method used

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  • A kind of lactobacillus plantarum and application thereof
  • A kind of lactobacillus plantarum and application thereof
  • A kind of lactobacillus plantarum and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The screening of embodiment 1 acid resistance and bile salt lactobacillus plantarum

[0048] Dilute 100 µL of freshly collected healthy human intestinal flora suspension with a 10-fold gradient, spread it on MRS agar medium, put it into an anaerobic culture tank, and use ternary mixed gas (80% nitrogen, 10% hydrogen) and carbon dioxide 10%) to replace the gas in the tank so that the final oxygen concentration in the culture tank was ≤5%, and the culture tank was cultivated in a 37°C incubator for 48 hours. Select independent, bright, and full-bodied colonies, and use Matrix-assisted laser desorption / ionization time–of-flight mass spectrometry (MALDI-TOF MS, referred to as flight mass spectrometry) technology for high-throughput bacterial colonies. a preliminary identification.

[0049] The colonies initially identified as Lactobacillus plantarum by MALDI-TOF MS equipment were picked into liquid MRS medium and cultured at 37°C for 12 hours, and then placed on MRS agar m...

Embodiment 2

[0054] Example 2 Preservation of bacterial strains

[0055] After the screened Lactobacillus plantarum NKK20 was streaked on the MRS agar medium for 48 hours, a single colony was selected and placed in a 10ml liquid medium, which was MRS liquid containing 4.5% soybean lecithin and 2% calcium carbonate Medium. Place the culture medium at a constant temperature of 37°C for 12h-16h, and measure the OD of the culture medium 600 When the value is greater than or equal to 1.2, add glycerol solution to the culture medium at a volume ratio of 1:1. The concentration of the glycerol solution is 30%-40%. Pre-cool at 4°C for 2 hours, then place at -20°C for 4 hours, and finally transfer to -80°C refrigerator or store in liquid nitrogen.

Embodiment 3

[0056] Example 3 Intestinal Mucosa Adhesion Experiment

[0057] Lactobacillus plantarum NKK20, Lactobacillus plantarum 229v and Lactobacillus plantarum LP-115 were selected for intestinal mucosal adhesion experiment.

[0058] After each strain was cultured at 37°C for 3 hours, scanning electron microscope analysis showed that all the strains adhered to the intestinal mucosa, and the adhesion rate of Lactobacillus plantarum NKK20 to the intestinal mucosa was basically the same as that of LP-115, about 81%. significantly higher than Lactobacillus plantarum 229v, and the adhesion rate of each strain to the intestinal mucosa is shown in Table 2.

[0059] Scanning electron microscope (SEM) image of intestinal mucoadhesive Lactobacillus plantarum NKK20, see Figure 4 .

[0060]

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Abstract

The present invention relates to a plant Lactobacillus ( Lactobacillus plantarum ) NKK20 and its applications belong to the field of microorganisms. The strain is isolated from the healthy human intestine and has been preserved in the China Center for Type Culture Collection with the preservation number CCTCC NO: M2020596. The Lactobacillus plantarum strain has strong acid resistance, bile salt resistance and intestinal mucosal cell adhesion; it can significantly inhibit pathogenic bacteria such as Salmonella enteritidis and Staphylococcus aureus, and has the function of inhibiting gastrointestinal pathogenic bacteria; it can significantly inhibit gastrointestinal pathogens in yogurt fermentation. Reducing the cholesterol content in milk can be applied in the fields of medicine, functional food and food additives, and is of great significance and value to human health.

Description

technical field [0001] The invention belongs to the field of microorganisms, in particular to a plant lactobacillus, and also relates to the application of the strain. Background technique [0002] Gastrointestinal diseases mainly refer to general inflammatory gastrointestinal diseases (acute and chronic gastritis, acute and chronic appendicitis, etc.), peptic ulcer, gastric cancer, esophageal cancer, colorectal cancer and irritable bowel syndrome. Nowadays, the incidence of gastrointestinal diseases is high. In my country, there is a saying that "nine out of ten people have stomach problems". The long course of disease, difficult treatment, and repeated attacks are all characteristics of gastrointestinal diseases. Research by medical experts has found that the cause of gastrointestinal diseases is the imbalance between protective factors and attacking factors in the gastric mucosa, resulting in weaker protective factors in the gastrointestinal mucosa than attacking factors,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/04C12N11/10A23L33/135A23C9/123A61K35/747A61P31/04C12R1/25
CPCC12N1/20C12N1/04C12N11/10A23L33/135A23C9/1234A61K35/747A61P31/04A23V2002/00A23V2400/169A23V2200/3262A23V2200/3204
Inventor 贾长生汪智婷王信超李珮铷
Owner 天益健康科学研究院(镇江)有限公司
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