Lactobacillus rhamnosus exopolysaccharide, and preparation method and application thereof

A technology of Lactobacillus rhamnosus and extracellular polysaccharide, applied in the field of microorganisms, can solve the problems of little research and the application prospect of probiotics in type 2 diabetes has not been widely explored, etc., and achieves a wide range of sources and good in vitro hypoglycemic activity. , Good in vitro hypoglycemic effect

Inactive Publication Date: 2021-04-16
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most data on the control of diabetes by probiotics are obtained directly from animal experiments, there are few studies on the specific mechanism of probiotics to regulate blood sugar, and the application prospects of probiotics in type 2 diabetes have not been widely explored
However, there is no report that exopolysaccharides extracted from probiotics with obvious α-glucosidase inhibitory activity also have similar in vitro hypoglycemic ability

Method used

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  • Lactobacillus rhamnosus exopolysaccharide, and preparation method and application thereof
  • Lactobacillus rhamnosus exopolysaccharide, and preparation method and application thereof
  • Lactobacillus rhamnosus exopolysaccharide, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: A screening method for a strain of Lactobacillus rhamnosus producing exopolysaccharides

[0058] The exopolysaccharide-producing Lactobacillus rhamnosus strains in this study were obtained from Xiangmanlou brand yoghurt in South China, and the culture was serially diluted with 0.85% normal saline (10 -1 -10 -7 ), and spread evenly on the MRS solid medium plate with a disposable spreader. The components of the MRS broth medium are (g L -1 ): Glucose (20), Tween 80 (1.08), MnSO 4 4H 2 O(0.05), MgSO 4 ·7H 2 O (0.2), Sodium Acetate Anhydrous (5), Triammonium Citrate (2), Casein (10), Beef Extract (10), Yeast Extract (4), K 2 HPO 4 (2) (pH5.7±0.2). Solid medium contains 1.5% agar. All materials were sterilized at 121°C for 20 minutes before use.

[0059] After culturing under anaerobic conditions at 37°C for 2 days, the strains with typical colony morphology were selected for Gram staining observation.

Embodiment 2

[0060] Example 2: Identification of the above-mentioned Lactobacillus rhamnosus strains producing exopolysaccharides

[0061] The Lactobacillus rhamnosus strain producing exopolysaccharide of the present invention has the following characteristics:

[0062] The single bacterium colony of bacterial strain described in the present invention on MRS solid culture medium is circular, smooth surface, milky white, and edge is neat, has drawing phenotype, and Gram's staining is purple-blue (such as figure 1 , as shown in 2).

[0063] The exopolysaccharide-producing Lactobacillus rhamnosus strain obtained through the above screening was tentatively named LB1lac10. In order to further determine the species of the strain, the genome of the strain was extracted by enzymatic hydrolysis, and the total DNA of the strain was extracted according to the steps of the Generay Biotech Bacteria Genomic DNA Extraction Kit (Generay Biotech Bacteria Genomic DNA Extraction Kit), and the sample was use...

Embodiment 3

[0066] Example 3: In vitro hypoglycemic ability of the above-mentioned Lactobacillus rhamnosus strain producing extracellular polysaccharides

[0067] The in vitro hypoglycemic ability of the above-mentioned Lactobacillus rhamnosus strain producing extracellular polysaccharides comprises the following steps:

[0068] Step 1: Inoculate LB1lac10 frozen at -80°C into liquid MRS broth medium and incubate at 37°C for 18 hours under anaerobic conditions. Then the second generation was activated for subsequent experiments, and the cultured LB1lac10 was centrifuged at 6000 rpm for 10 minutes at 4°C.

[0069] Viable bacterial suspension: After centrifugation, wash the LB1lac10 culture solution twice with phosphate buffered saline (PBS, 0.1M, pH=6.8), and adjust the strain concentration to 1×10 9 CFU / mL.

[0070] Inactivated bacteria suspension: Live bacteria (1×10 9 CFU / mL) was boiled at 100°C for 20 minutes.

[0071] Step 2: First, add 25 μL of 20 mmol / L PNPG (4-nitrophenyl-D-gluc...

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Abstract

The invention discloses a lactobacillus rhamnosus exopolysaccharide, and a preparation method and application thereof, and belongs to the technical field of microorganisms. According to the invention, a lactobacillus rhamnosus strain separated from Xiangshuangpao yoghurt is used as an original strain to produce exopolysaccharide, the exopolysaccharide is a uniform component, and the molecular weight of the exopolysaccharide is 88650 Da; and the lactobacillus rhamnosus exopolysaccharide comprises mannose, glucuronic acid, glucose, xylose, galactose and arabinose. The exopolysaccharide has a good in-vitro hypoglycemic effect, has good in-vitro alpha-glucosidase inhibition capacity through detection, and is simple in process, wide in market prospect, stable in thermodynamic property and suitable for large-scale production. Therefore, the polysaccharide is a potential active ingredient for preventing and treating diabetes mellitus. Besides, as a fermentation product of probiotics, compared with common chemical medicines in the field of blood sugar reduction, the fermentation product has the advantages of simple and easily available active ingredients, low price, no side effect and the like, and has great application value.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to an exopolysaccharide of Lactobacillus rhamnosus and a preparation method and application thereof. Background technique [0002] With the improvement of people's living standards, the incidence of diabetes is getting higher and higher due to excessive nutritional intake. Type 2 diabetes mellitus (T2 DM) is one of the most common chronic diseases in patients with abnormally high blood glucose levels. Due to the complex pathogenesis of this disease and many influencing factors, there is no effective prevention and control measures at present. [0003] Clinically used drugs for diabetes mainly include acarbose, voglibose and miglitol. Among these diabetes drugs, acarbose is an α-glucosidase inhibitor that can compete with oligosaccharides on the side of small intestinal epithelial cells to reversibly bind to α-glucosidase. Its activity can inhibit the hydrolysis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12N1/20C08B37/00A61K31/7088A61P3/10C12R1/225
Inventor 王家彬潘力王斌
Owner SOUTH CHINA UNIV OF TECH
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