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Lactobacillus plantarum lipoteichoic acid and application thereof in inhibition of amyloid protein aggregation

A technology of Lactobacillus plantarum and amyloid protein, applied in the fields of application, food science, drug combination, etc., can solve the problems that hinder the prevention and relief of probiotics, the mechanism of action is not clear, and the types of probiotics are scarce, so as to achieve high safety, Effect of inhibiting Aβ42 aggregation

Pending Publication Date: 2022-03-01
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is a shortage of probiotics known to prevent and treat AD. The complexity of intestinal flora and their metabolites makes research more difficult. The mechanism of using intestinal microecology to improve AD is still unclear. These are obstacles to probiotics. Key scientific issues in bacterial prevention and mitigation of AD research

Method used

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  • Lactobacillus plantarum lipoteichoic acid and application thereof in inhibition of amyloid protein aggregation
  • Lactobacillus plantarum lipoteichoic acid and application thereof in inhibition of amyloid protein aggregation
  • Lactobacillus plantarum lipoteichoic acid and application thereof in inhibition of amyloid protein aggregation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 : Isolation, extraction and purification of Lactobacillus plantarum MA2 lipoteichoic acid (lpLTA)

[0028] (1) Inoculate Lactobacillus plantarum MA2 in a biological safety cabinet, and cultivate it to the exponential growth phase under corresponding conditions;

[0029] (2) Take the bacterial liquid in a centrifuge cup, centrifuge at 4°C and 7000rpm for 20min, collect the bacterial cells, and weigh the wet weight of the bacterial cells;

[0030] (3) Resuspend the wet bacteria in 0.1M sodium citrate buffer solution with pH 4.7 at a ratio of 1:1 (w / v). 1s, off 1s, total working time 30min;

[0031] (4) Add an equal volume of n-butanol to the mixed bacterial suspension and mix thoroughly, and stir on a magnetic stirrer for 30 minutes, centrifuge at 4°C and 7000 rpm for 30 minutes, and gently collect the lower aqueous phase;

[0032] (5) The aqueous phase is subjected to rotary evaporation and dialysis, and the molecular weight cut-off of the dialysis bag is 2...

Embodiment 2

[0042] Example 2 : Structural identification of lipoteichoic acid (lpLTA) from Lactobacillus plantarum MA2

[0043] (1) Infrared spectral analysis

[0044] Weigh an appropriate amount of lpLTA powder, grind it with potassium bromide at a mass ratio of 1:100, press the mixed powder into tablets, and then use an infrared spectrometer to analyze the functional groups of lpLTA, the results are as follows figure 1 shown.

[0045] refer to figure 1 , Band 3419.95cm -1 Characterization - NH group, band 2929.31cm -1 Characterization-CH 2 、-CH 3 Group, band 1651.56cm -1 Characterization - C=O group, band 1547.32cm -1 Characterization - C=H group, band 1379.09cm -1 Characterization - P=O group, band 1231.18cm -1 Characterization of sugar residue groups, band 1038.81cm -1 Characterization of the -C-O group. This result is basically consistent with the characteristic functional groups of the general structure of lipoteichoic acid. At the same time, the infrared spectrum resul...

Embodiment 3

[0049] Example 3 : The aggregation inhibitory effect of different concentrations of Lactobacillus plantarum MA2 lipoteichoic acid (lpLTA) on Aβ42 was detected by Thioflavine T (ThT) method

[0050] Prepare an Aβ42 solution with a final concentration of 25 μM, add 0.25, 0.50, 1.00, 2.00 mg / mL lpLTA to it, and add 25 μM ThT dye, mix thoroughly, place in a microplate reader, and culture continuously at 37 ° C, every 1 h Detect the fluorescence of the sample. The detection condition is that the excitation light wavelength is 440nm, and the emission light wavelength is 480nm. ThT fluorescence results are as follows: image 3 shown.

[0051] refer to image 3 , the fluorescence intensity of Aβ42 itself gradually increased with time, and basically reached the plateau after ~24h, and after adding 0.25mg / mL lpTLA, the fluorescence intensity of Aβ42 solution also gradually increased with time, but reached the plateau after ~12h, and the maximum fluorescence intensity It is about 60%...

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Abstract

The invention provides lactobacillus plantarum lipoteichoic acid and application of the lactobacillus plantarum lipoteichoic acid in inhibition of amyloid protein aggregation. The main chain of the lactobacillus plantarum lipoteichoic acid contains glycerol, and the side chain of the lactobacillus plantarum lipoteichoic acid contains a large amount of N-acetyl glucosamine and alanine modification. The lipoteichoic acid can effectively inhibit aggregation of beta-amyloid protein 1-42 (Abeta42), is a potential Abeta42 aggregation inhibitor, and can be applied to development of drugs, health care products or functional food for preventing and / or improving Alzheimer's disease.

Description

technical field [0001] The invention belongs to the technical field of medicine, health care products or food, and in particular relates to the separation, extraction and structure identification of a new type of Lactobacillus plantarum lipoteichoic acid and its application in inhibiting the aggregation of amyloid protein, in particular to the application of Lactobacillus plantarum as a plant Lactobacillus MA2, the use of its lipoteichoic acid as a β-amyloid 1-42 (Aβ42) aggregation inhibitor in the preparation of drugs for preventing and / or improving Alzheimer's disease, health products and functional foods. Background technique [0002] Aggregation and misfolding of amyloid in cells and tissues to form toxic intermediates and amyloid fibrils may lead to various biological dysfunctions. These aggregation intermediates and fibers are speculated to be related to some neurodegenerative diseases and other conditions, such as Alzheimer's disease (AD), Parkinson's disease (Parkins...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/747A61P25/28C12P7/64A23L33/135
CPCA61K35/747A61P25/28A23L33/135A23V2002/00A23V2200/322A23V2250/206Y02A50/30
Inventor 贾龙刚王远望王艳萍耿伟涛王金菊
Owner TIANJIN UNIV OF SCI & TECH
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