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Preparation method of functional lactobacillus acidophilus peptidoglycan and anti-inflammatory activity use of functional lactobacillus acidophilus peptidoglycan

A technology of Lactobacillus acidophilus and peptidoglycan is applied in the field of preparation of functional Lactobacillus acidophilus peptidoglycan, which can solve the problem that the anti-inflammatory activity of Lactobacillus acidophilus peptidoglycan has not been studied.

Active Publication Date: 2014-08-13
厦门赋源康实业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no relevant reports on the preparation method and anti-inflammatory activity of Lactobacillus acidophilus peptidoglycan, and there are no functional foods related to Lactobacillus acidophilus peptidoglycan on the market.

Method used

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  • Preparation method of functional lactobacillus acidophilus peptidoglycan and anti-inflammatory activity use of functional lactobacillus acidophilus peptidoglycan
  • Preparation method of functional lactobacillus acidophilus peptidoglycan and anti-inflammatory activity use of functional lactobacillus acidophilus peptidoglycan
  • Preparation method of functional lactobacillus acidophilus peptidoglycan and anti-inflammatory activity use of functional lactobacillus acidophilus peptidoglycan

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Experimental program
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specific Embodiment 1

[0019] Preparation of Lactobacillus acidophilus peptidoglycan

[0020] Lactobacillus acidophilus ( Lactobacillus acidophilus ) The preservation number is CGMCC No.8920, inoculate the inoculation amount of 2.0-4.0% by volume, inoculate on MRS medium, and after anaerobic culture at 37°C for 18-24h, centrifuge at 4000-8000g for 15-20min to remove bacteria Then take 1 g of the precipitated bacteria and add it to the SDS solution with a mass fraction of 4%, and pyrolyze it at 100°C for 30 minutes to remove the protein component; take the precipitate and add a solution of trichloroacetic acid with a mass fraction of 10% to the precipitate , to remove teichoic acid; take the precipitate, dialyze the precipitate overnight at 4°C for desalination, pre-freeze the precipitate at -80°C for 6 hours, and quickly put it into a vacuum freeze-drying device for drying at 80-120Pa for 24 hours to obtain The powder is Lactobacillus acidophilus peptidoglycan. Samples were stored in a -40°C freez...

specific Embodiment 2

[0026] The Lactobacillus acidophilus peptidoglycan prepared in Example 1 above was subjected to mass spectrometry (MALDI-TOF / TOF MS) detection and analysis; the Lactobacillus acidophilus peptidoglycan powder was subjected to amino acid analysis and infrared (RI) detection.

[0027] 1. Molecular weight and substructure of peptidoglycan

[0028] Mix 1 microliter of Lactobacillus acidophilus peptidoglycan solution with a concentration of 1 mg / ml and 1 microliter of 50 mg / ml 2,5-dihydroxybenzoic acid. After mixing evenly, take 1 microliter of the mixture and load it. Perform mass spectrometry. The instrument model is FLEX-PC, and the type is ultraflex TOF / TOF. The molecular weight of peptidoglycan detected by this protocol is 875.260 Da, and the specific analysis is shown in Table 1 below.

[0029] Table 1 List of peptidoglycan ion fragments

[0030]

[0031] 2. Analysis of amino acid content

[0032] Use the A200 Amino Nova amino acid analyzer to analyze the composition an...

specific Embodiment 3

[0038] Anti-inflammatory activity analysis of the Lactobacillus acidophilus peptidoglycan prepared in the above-mentioned embodiment 1

[0039] (1) Using Lactobacillus acidophilus peptidoglycan to regulate the inflammation model of LPS-induced macrophage Raw 264.7, analyze the expression changes of iNOS and COX-2 in the final product, and analyze whether peptidoglycan has a good inhibitory effect Inflammatory substance releasing activity.

[0040] Raw 264.7 cells were cultured in a six-well plate. In the normal group, the adhesion rate reached 60-80%. The RAW 264.7 macrophages were induced by adding LPS at a final concentration of 100ng / ml, and the cytoplasmic protein was extracted after 18 h of continuous culture; the peptidoglycan repair group added final LPS with a concentration of 100ng / ml and peptidoglycan with a final concentration of 25μg / ml, 50μg / ml, 100μg / ml, and 200μg / ml were sequentially cultured for 18 hours to extract cytoplasmic proteins.

[0041] Different gr...

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Abstract

The invention discloses a preparation method of a functional lactobacillus acidophilus peptidoglycan and an anti-inflammatory activity use of the functional lactobacillus acidophilus peptidoglycan. The preparation method comprises the following steps of inoculating a MRS culture medium with lactobacillus acidophilus having an accession number of CGMCC No. 8920 according to an inoculation ratio of 2.0-4.0% by volume, carrying out anaerobic cultivation at a temperature of 37 DEG C for 18-24h, carrying out centrifugation under a centrifugation force of 4000-8000g for 15-20min so that a fermentation broth is removed, adding 4wt% of a SDS solution into the precipitated acidophilus peptidoglycan, carrying out high-temperature pyrolysis at a temperature of 100 DEG C for 30min to remove a protein component, adding 10wt% of a trichloroacetic acid solution into the precipitates to remove teichoic acid, carrying out overnight dialysis desalination on the precipitates at a temperature of 4 DEG C, carrying out pre-refrigeration on the precipitates at a temperature of -80 DEG C for 6h, fast putting the refrigerated precipitates into a drying device, and carrying out drying treatment under the pressure of 80-120Pa for 24h to obtain a powdery product. The functional lactobacillus acidophilus peptidoglycan has good inhibition and conditioning effects on LPS-caused inflammation.

Description

technical field [0001] The invention relates to a preparation method of peptidoglycan, in particular to a preparation method of functional Lactobacillus acidophilus peptidoglycan and its anti-inflammatory activity application. Background technique [0002] Lactic acid bacteria, as an important type of colonizing intestinal probiotics, are widely used in fermented food processing, especially yogurt, cheese, pickles, etc. The close relationship between intestinal lactic acid bacteria and the health of the host has been expounded by more and more microbiologists at home and abroad, especially in terms of inducing the release of immune regulatory factors to stimulate the immune system to exert its biological effects such as anti-infection and anti-tumor. In terms of the immunomodulatory activity of lactic acid bacteria on the host, recent studies have found that there is no significant difference between some live bacteria and dead bacteria. Muic acid (LTA) and surface protein ...

Claims

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Application Information

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IPC IPC(8): C12P19/04A61K31/715A61P29/00A23L1/29C12R1/23A23L33/00
CPCY02A50/30
Inventor 潘道东吴振孙杨赢曾小群曹锦轩李桦
Owner 厦门赋源康实业有限公司
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