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Lactobacillus plantarum exopolysaccharide and preparation method thereof

A technology of Lactobacillus plantarum and exopolysaccharides, which is applied in the field of Lactobacillus plantarum exopolysaccharides and its preparation, can solve the problems of lack of Lactobacillus plantarum exopolysaccharides and difficult preparation of Lactobacillus plantarum exopolysaccharides, and achieve purity and yield rate-boosting effect

Inactive Publication Date: 2016-08-03
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention provides an exopolysaccharide of Lactobacillus plantarum to solve the technical problem of lacking the exopolysaccharide of Lactobacillus plantarum in the prior art
[0007] The present invention also provides a method for preparing the above-mentioned exopolysaccharide of Lactobacillus plantarum to solve the technical problem that the above-mentioned exopolysaccharide of Lactobacillus plantarum is difficult to prepare in the prior art

Method used

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  • Lactobacillus plantarum exopolysaccharide and preparation method thereof
  • Lactobacillus plantarum exopolysaccharide and preparation method thereof
  • Lactobacillus plantarum exopolysaccharide and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0049] 1.1 Preparation of exopolysaccharide of Lactobacillus plantarum

[0050] (1) Take Lactobacillus plantarum with the preservation number CCTCCM2014170, ferment it in MRS medium, the fermentation time is 24h, anaerobic culture at 37°C;

[0051] (2) Use 2 times the volume of absolute ethanol to precipitate the supernatant of the fermentation broth for 48 hours, centrifuge at 12,000×g for 10 minutes, discard the supernatant, and dialyze the precipitate with a dialysis bag with a molecular weight cut-off of 8,000-14,000 Da for 3 days, and change the ultrapure water twice a day. Freeze-drying to obtain crude exopolysaccharide;

[0052] (3) Use 50mM Tris-HCl, 10mM MgSO 4 ·7H 2 O dissolve the crude polysaccharide with a final concentration of 5 mg / mL, and use nuclease DNAsetype-I with a final concentration of 2.5 μg / mL to hydrolyze the nucleic acid at 37°C for 6 hours;

[0053] (4) The hydrolyzed crude polysaccharide was hydrolyzed with protease Pronase E at a final concentra...

Embodiment 2

[0071] The exopolysaccharide of Lactobacillus plantarum is composed of xylose and galactose connected by glycosidic bonds, wherein the molar content of galactose is 98%. The number of elution peaks of the exopolysaccharide of Lactobacillus plantarum through space exclusion chromatography is one.

[0072] The preparation method of the above-mentioned exopolysaccharide of Lactobacillus plantarum comprises the following steps:

[0073] 1) Take the Lactobacillus plantarum fermentation liquid, separate the solid from the liquid and get the supernatant;

[0074] 2) adding ethanol to the supernatant in step 1), ethanol precipitation, solid-liquid separation to take the precipitate, dialysis with a dialysis bag with a molecular weight cut-off of 8KDa, and drying the contents of the dialysis bag to obtain crude polysaccharide;

[0075] 3) To contain 40mM Tris-HCl, 5mM MgSO 4 ·7H 2 The solution of O is used as the crude polysaccharide solution, dissolves the crude polysaccharide desc...

Embodiment 3

[0081] The exopolysaccharide of Lactobacillus plantarum is composed of xylose and galactose connected by glycosidic bonds, wherein the molar content of galactose is 98.3%.

[0082] The preparation method of the above-mentioned exopolysaccharide of Lactobacillus plantarum comprises the following steps:

[0083] 1) Take the Lactobacillus plantarum fermentation liquid, separate the solid from the liquid and get the supernatant;

[0084] 2) adding ethanol to the supernatant in step 1), ethanol precipitation, solid-liquid separation to take the precipitate, dialysis with a dialysis bag with a molecular weight cut-off of 14KDa, and drying the contents of the dialysis bag to obtain crude polysaccharide;

[0085] 3) to contain 60mM Tris-HCl, 15mM MgSO 4 ·7H 2 The solution of O is used as the crude polysaccharide solution, dissolves the crude polysaccharide described in step 2), and then adds nuclease DNAsetype-I to a final concentration of 3 μg / mL for hydrolysis;

[0086] 4) Then a...

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Abstract

The invention provides a lactobacillus plantarum exopolysaccharide. According to the exopolysaccharide, sugar units in the molecular structure are only xylose and galactose, the galactose accounts for 98%, and the exopolysaccharide belongs to a lactobacillus plantarum exopolysaccharide of a brand new structure. Meanwhile, the invention provides a method for preparing the exopolysaccharide. The method comprises the following steps: precipitating polysaccharides in lactobacillus plantarum fermentation solution by ethanol, dialyzing to remove small molecules, removing nucleic acid and protein impurities by the enzymolysis methods; precipitating impurity proteins by trichloroacetic acid, performing solid-liquid separation, taking the supernatant, and performing dialysis, thereby obtaining the purified target polysaccharide. According to the method, due to the property of the target polysaccharide and impurity component design process parameters of lactobacillus plantarum fermentation solution, the temperature and pH conditions are mild, purification methods capable of possibly breaking polysaccharide cross-linked bonds such as ultrafiltration are avoided, and integrity of polysaccharide glycosidic bonds and secondary structures is effectively guaranteed. The polysaccharide prepared by the method is high in purity and yield and lays a foundation for subsequent performance analysis and industrial application.

Description

technical field [0001] The invention relates to the technical field of biological separation engineering, and further relates to the separation and purification of microbial fermentation products, in particular to a plant Lactobacillus exopolysaccharide and a preparation method thereof. Background technique [0002] Lactic acid bacteria exopolysaccharide (Exopolysaccharide, EPS), generally refers to the carbohydrate compound secreted outside the cell during the metabolism of lactic acid bacteria. Since the exopolysaccharide of lactic acid bacteria has good rheology and can give fermented food a good taste and flavor, it can be used as a gelling agent, stabilizer, preservative and emulsifier in the food industry. It has been widely used in dairy products, Production of beverages, pastries, candies, ice cream and other foods. In addition, studies have shown that EPS has the functions of anti-pathogenic bacteria, anti-tumor, lowering cholesterol, delaying aging, regulating the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12R1/25
CPCC12P19/04
Inventor 魏华张志鸿陶雪莹徐锋夏慧玲许恒毅万翠香
Owner NANCHANG UNIV
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