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Method for extracting peptidoglycan from lactobacillus

A technology of peptidoglycan and lactobacillus, which is applied in the field of preparation of peptidoglycan extracted from Lactobacillus helveticus by physical and chemical methods, can solve the problems such as the lack of in-depth research on the immunological activity of PGN and the failure to reach a certain consensus, and the yield can be achieved. High, low cost and practical effect

Inactive Publication Date: 2021-06-25
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The probiotic activity of lactic acid bacteria has been accepted by the majority of consumers, and its anti-oxidation and function of improving the balance of intestinal flora have also been confirmed by many researchers. A certain consensus has not been reached, and no in-depth research has been conducted on the way through which PGN exerts its immunological activity

Method used

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  • Method for extracting peptidoglycan from lactobacillus
  • Method for extracting peptidoglycan from lactobacillus
  • Method for extracting peptidoglycan from lactobacillus

Examples

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specific Embodiment 1

[0031] Specific embodiment 1: the preparation method of Lactobacillus helveticus peptidoglycan:

[0032] (1) Preparation of bacteria

[0033] Lactobacillus helveticus was activated twice, and after continuous culture at 37°C for 24 hours to reach a stable period, the bacterial cells in the fermentation broth were centrifuged at 6000×g for 25 minutes to obtain them, and centrifuged and washed with physiological saline for 3 times.

[0034] (2) Lysis of bacteria

[0035] The obtained thallus was boiled in 4% SDS solution for 30 minutes to promote the lysis of the thallus and dissolution of intracellular components. Then the temperature was lowered at 4°C for 5 minutes, and the bacterial pellet was centrifuged at 6000×g for 25 minutes at room temperature. After removing the supernatant, it was washed with deionized water until the trace of SDS was reached.

[0036] (3) Increased cell wall permeability of bacteria

[0037] Ultrasonic crushing was used for the bacterial cell pre...

specific Embodiment 2

[0047] The extraction rate of peptidoglycan is calculated according to the following formula, and the extraction rate of peptidoglycan obtained in this example is 17%.

[0048] Extraction rate (%)=(peptidoglycan dry weight / cell dry weight)×100

specific Embodiment 3

[0050] The purity of peptidoglycan was verified by HPLC method (e.g. figure 2). Agilent analysis system was adopted, chromatographic column: TSK-GELG4000 PWXL chromatographic column (300×7.8mm), detection conditions: column temperature 37°C, flow rate 0.6ml / min, mobile phase: deionized water, PGN sample volume 20μl (1mg / ml), detector: UV225nm ultraviolet detector. The result is as figure 2 As shown, the peptidoglycan derived from the cell wall of Lactobacillus helveticus MB2-1 peaked at a time of 8.738 minutes. The purity of the peptidoglycan obtained in this example is 100%.

[0051] Purity (%)=(peak area of ​​peptidoglycan / total peak area of ​​detection sample)×100

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Abstract

The invention relates to a method for extracting peptidoglycan from Lactobacillus helveticus MB2-1. The method is characterized in that the permeability of cell walls is increased by physical and chemical methods, the physical method adopts an ultrasonic crushing method, and the chemical method adopts a trichloroacetic acid method to extract teichoic acid so as to increase the permeability of cell membranes. Fat substances on cell membranes are removed through degreasing of an organic reagent, protein substances in cells are removed through composite enzymatic hydrolysate, finally, impurities are removed through centrifugal cleaning, and freeze-drying is conducted to obtain peptidoglycan. The preparation method has the advantages that the extracted complete peptidoglycan with the probiotic effect is high in practicability, simple to operate, short in period, simple, convenient and reasonable in step, low in cost, high in yield and suitable for large-scale production.

Description

technical field [0001] The invention relates to a preparation method of peptidoglycan, in particular to a preparation method for extracting peptidoglycan of Lactobacillus helveticus by a physical and chemical method. Background technique [0002] As an important intestinal probiotic, lactic acid bacteria are one of the most important physiological flora in the intestinal tract of humans and animals. Lactic acid bacteria are not only non-toxic, harmless, and have no side effects, but also have various health functions, such as regulating intestinal flora, maintaining intestinal microecological balance, alleviating lactose intolerance, lowering cholesterol, lowering blood pressure, enhancing immunity, Oxidation, anti-tumor and anti-mutation, etc. Studies have shown that the probiotic effect of lactic acid bacteria can be realized through its own primary metabolism, and it is also related to the structure of lactic acid bacteria itself, such as closely related to the peptidogl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00
CPCC08B37/0003
Inventor 李伟王凯旋芮昕董明盛张秋勤陈晓红
Owner NANJING AGRICULTURAL UNIVERSITY
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