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A strain of Lactobacillus plantarum nx-1 and its application in the preparation of hypoglycemic drugs

A Lactobacillus plantarum, hypoglycemic technology, applied in the field of microorganisms, can solve problems such as not necessarily suitable, probiotic strains relying on imports, lack of scientific research evidence for probiotic functions, etc. - Effects of glucosidase and alpha-amylase

Active Publication Date: 2022-04-01
广东南芯医疗科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many types of probiotic products in the domestic market, there is a lack of functional strains with independent intellectual property rights in my country
The probiotic strains used by domestic production enterprises have long relied on imports, and foreign strains may not be suitable for the intestinal physiological conditions of Chinese residents
In addition, the function of probiotics lacks strong scientific research evidence, which seriously affects the promotion of probiotics and their products

Method used

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  • A strain of Lactobacillus plantarum nx-1 and its application in the preparation of hypoglycemic drugs
  • A strain of Lactobacillus plantarum nx-1 and its application in the preparation of hypoglycemic drugs
  • A strain of Lactobacillus plantarum nx-1 and its application in the preparation of hypoglycemic drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Isolation, Identification and Preservation of Novel Lactobacillus plantarum NX-1

[0027] (1) Separation: After gradient dilution of the spicy cabbage fermentation liquid, inoculate them in TPY solid medium, MRS solid medium and BDS solid medium respectively, culture anaerobically at 37°C for 48 hours, pick a single colony on the plate and streak Pure colonies were isolated. The pure colonies on the plate were inoculated into MRS liquid medium, cultured anaerobically at 37°C for 12-16h, added 20% glycerol, and stored in a -80°C refrigerator.

[0028] (2) Morphological identification of bacterial strains: Gram-stained strains were observed under a microscope, Gram-positive bacteria were purple, and Gram-negative bacteria were red.

[0029] (3) Molecular biological identification of the strain: Genomic DNA was extracted from the obtained strain, and the full-length 16S rDNA fragment was amplified by PCR technology using 16S rDNA universal primers 27F and 1492R,...

Embodiment 2

[0035] The preparation of embodiment 2 novel Lactobacillus plantarum NX-1 fermentation supernatant (extracellular secretion), bacterium suspension (thalline), cell disruption supernatant (intracellular substance)

[0036] The new Lactobacillus plantarum NX-1 was activated and cultured and inoculated in MRS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermentation bacteria was adjusted to 10 9 CFU / mL, centrifuged at 4°C, 6000r / min for 10min to obtain the culture supernatant and cell pellet, the supernatant was filtered through a 0.22μm filter membrane to obtain the fermentation supernatant (extracellular secretion); the cell pellet was filtered through PBS twice After the first wash, the cells were resuspended with PBS, and the cell concentration was adjusted to 10 9 CFU / mL obtains bacterial suspension (thalline); Wash and resuspend concentration to be 10 9 The CFU / mL bacterial suspension was treated in an ice bath with an ultrasonic breaker, worki...

Embodiment 3

[0037] Example 3 Analysis of the inhibitory ability of novel Lactobacillus plantarum NX-1 to α-glucosidase activity

[0038] Mix 100 μL of 1 U / mL α-glucosidase (0.1M PBS, pH 6.8) with 50 μL of samples (non-inactivated and inactivated: fermentation supernatant, bacterial suspension, cell lysate supernatant) in 1.5 mL in a centrifuge tube. After pre-incubation for 10 min in a water bath at 37° C., 50 μL of 5 mM p-nitrophenyl-α-D-glucopyranoside (pNPG, 0.1 M PBS, pH 6.8) was added as a substrate. The enzymatic reaction was carried out in a water bath at 37°C for 30 min, and by adding 1 mL of 0.1M Na 2 CO 3 (distilled water) to terminate the reaction. 200 μL of the reaction solution was placed in a 96-well cell culture plate, and the α-glucosidase activity was determined by measuring the release of p-nitrophenol from pNPG at 405 nm. The solvent without sample was used as a control, the solvent without substrate was used as a blank, and acarbose was used as a positive control. ...

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Abstract

The invention discloses a strain of Lactobacillus plantarum NX‑1 and its application in the preparation of hypoglycemic drugs, belonging to the technical field of microorganisms. A strain of Lactobacillus plantarum NX-1 disclosed by the present invention has a preservation number of CGMCC No.20109. Lactobacillus plantarum NX‑1 inactivated and inactivated fermentation supernatants, bacterial suspensions, and supernatants of cell disruptions have strong inhibitory effects on α-glucosidase and α-amylase; and both can Significantly reduce the glucose content in the zebrafish hyperglycemia model. The Lactobacillus plantarum NX‑1 disclosed by the invention has great potential application prospects in the preparation of medicines for treating and / or preventing diabetes.

Description

technical field [0001] The invention relates to the technical field of microorganisms, and more specifically relates to a strain of Lactobacillus plantarum NX-1 and its application in the preparation of hypoglycemic drugs. Background technique [0002] Diabetes mellitus is a severe metabolic disease characterized by chronic hyperglycemia due to impaired insulin production (type 1 diabetes / T1DM) or insulin resistance (type 2 diabetes / T2DM). According to the statistics of the International Diabetes Federation (IDF), about 463 million adults aged 20-79 worldwide suffer from diabetes in 2019; it is estimated that by 2030, the number of diabetic patients will reach 578.4 million; by 2045, the number of diabetic patients will reach 700.2 million . The top three countries with the largest number of diabetic patients in 2019 are China, India and the United States, and the number of diabetic patients (20-79 years old) is 116.4 million, 77 million and 31 million respectively. In rec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/747A61P3/10C12R1/25
CPCA61K35/747A61P3/10C12R2001/25C12N1/205
Inventor 张召郑康帝赵笑笑
Owner 广东南芯医疗科技有限公司
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