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Preparation method of modified lactobacillus acidophilus peptidoglycan

A technology of Lactobacillus acidophilus and peptidoglycan, which is applied in the field of preparation of modified Lactobacillus acidophilus peptidoglycan, can solve the problem of unfavorable PG efficacy research and industrialization, large changes in PG molecular structure, difficult control of enzyme activity, etc. problem, to achieve the effect of improved solubility and antioxidant activity, short reaction time, and improved solubility

Active Publication Date: 2013-11-20
武汉益鼎天养生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, lysozyme is expensive, takes a long time, the enzyme activity is not easy to control, and the obtained PG molecular structure changes greatly, which is not conducive to the research and industrialization of PG efficacy.

Method used

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  • Preparation method of modified lactobacillus acidophilus peptidoglycan
  • Preparation method of modified lactobacillus acidophilus peptidoglycan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A preparation method of modified Lactobacillus acidophilus peptidoglycan, comprising the following steps:

[0025] (1) Preparation of wet cells

[0026] Put 3g of Lactobacillus acidophilus in a Erlenmeyer flask, heat and kill the bacteria in a water bath at 65°C for 40min, then add 10mL of 0.5% Triton X-100 (polyethylene glycol octylphenyl ether) solution, and bathe in a water bath at 85°C for 1h , and continuously stirred, take out the solution and put it in an ice bath to cool immediately, centrifuge at 12000×g for 10 min, discard the supernatant, wash the precipitate twice with double distilled water, and centrifuge again at 12000×g for 8-12 min Discard the supernatant, then wash with methanol-water (v:v=2:1) ​​and methanol in sequence to remove impurities in the precipitate to obtain wet cells;

[0027] (2) Enzymatic hydrolysis and wall breaking

[0028] Add the wet bacteria to 8mL enzyme solution Ⅰ, put it in a water bath at 37°C for 12 hours, then centrif...

Embodiment 2

[0037] With embodiment 1, its difference is:

[0038] In step (1): put Lactobacillus acidophilus in the Erlenmeyer flask, bathe in water at 60°C for 45 minutes, then add 0.5% Triton X-100 solution, bathe in water at 80°C for 1.5h, and keep stirring continuously, take out the solution and put it in an ice bath Cool immediately, centrifuge at 10,000×g for 12 min, discard the supernatant, wash the precipitate twice with double distilled water, centrifuge again at 10,000×g for 12 min, and discard the supernatant.

[0039] In step (2): add wet bacteria to enzyme solution Ⅰ, place in 35°C water bath for 14 h, then centrifuge at 4°C, 10,000×g for 50 min, discard the supernatant, take the precipitate and add to enzyme solution Ⅱ, place in 35°C water bath for 14 h h, then centrifuge at 4°C, 10,000×g for 50 minutes, discard the supernatant, take the precipitate and add it to enzyme solution III, treat it in a water bath at 35°C for 20 minutes, then centrifuge at 4°C, 10,000×g for 50 m...

Embodiment 3

[0044] With embodiment 1, its difference is:

[0045] In step (1): put Lactobacillus acidophilus in the Erlenmeyer flask, bathe in water at 70°C for 35 minutes, then add 0.5% Triton X-100 solution, bathe in water at 90°C for 0.5h, and keep stirring continuously, take out the solution and put it in an ice bath Cool immediately, centrifuge at 15,000×g for 8 min, discard the supernatant, wash the precipitate twice with double distilled water, and centrifuge again at 15,000×g for 8 min, then discard the supernatant.

[0046] In step (2): add wet bacteria to enzyme solution Ⅰ, bathe in water at 40°C for 10 hours, then centrifuge at 4°C and 15,000×g for 30 minutes, discard the supernatant, take the precipitate and add it to enzyme solution Ⅱ, bathe in water at 40°C for 10 hours, Then centrifuge at 4°C, 15,000×g for 30 minutes, discard the supernatant, take the precipitate and add it to enzyme solution III, treat it in a water bath at 40°C for 20 minutes, then centrifuge at 4°C, 15...

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Abstract

The invention discloses a preparation method of modified lactobacillus acidophilus peptidoglycan. The preparation method is characterized by comprising the following steps of: sterilizing lactobacillus acidophilus into a water bath, adding the lactobacillus acidophilus into a TritonX-100 solution to react, and then, centrifugally washing to remove impurities in sediments to obtain a wet thallus; centrifuging after adding the wet thallus into an enzyme solution I to react, centrifuging after placing the sediments into an enzyme solution II to react, and centrifuging after adding the sediments into an enzyme solution III to react to obtain an enzymolysis wall broken thallus; washing and degreasing the enzymolysis wall broken thallus, and then, adding the enzymolysis wall broken thallus into an enzyme solution IV to remove proteins; dialyzing and salting out the degreased and protein-removed sediments, centrifuging and dialyzing the sediments, and then, freezing and drying to obtain the lactobacillus acidophilus peptidoglycan; and adding the lactobacillus acidophilus peptidoglycan into tetramentylniperidine to react with NaBr and NaClO to obtain an oxidized and modified lactobacillus acidophilus peptidoglycan product. The preparation method has the advantages of simplicity in operation, short reaction time and remarkable improvement of solubility and antioxidant activity.

Description

technical field [0001] The invention relates to a preparation method of peptidoglycan, in particular to a preparation method of modified Lactobacillus acidophilus peptidoglycan. Background technique [0002] As an important intestinal probiotic, lactic acid bacteria play an important role in regulating the intestinal microbial flora, enhancing the body's immunity and preventing diseases such as tumors. Lactobacillus acidophilus is an important member of the lactic acid bacteria family, which plays an important role in adjusting the balance of intestinal flora, reducing serum cholesterol levels, stimulating the immune system, and improving immune function. The principle of Lactobacillus acidophilus' immune regulation is that its peptidoglycan can regulate the immune function of the host by inducing the release of various cytokines. [0003] Peptidoglycan (PG) is one of the main components of the bacterial cell wall, and its monomer is composed of a tetrapeptide tail and a di...

Claims

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Application Information

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IPC IPC(8): C08B37/00
Inventor 潘道东杨媛曾小群曹锦轩孙扬赢
Owner 武汉益鼎天养生物科技有限公司
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