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A strain of Lactobacillus plantarum reducing the antigenicity of milk β-lactoglobulin and its application

A technology of Lactobacillus plantarum and lactoglobulin, applied in the field of Lactobacillus plantarum AHQ-14, to achieve the effect of high adhesion ability and high survival rate of intestinal cells

Active Publication Date: 2022-06-21
DALIAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no public report on the use of Lactobacillus plantarum AHQ-14 to degrade milk allergens

Method used

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  • A strain of Lactobacillus plantarum reducing the antigenicity of milk β-lactoglobulin and its application
  • A strain of Lactobacillus plantarum reducing the antigenicity of milk β-lactoglobulin and its application
  • A strain of Lactobacillus plantarum reducing the antigenicity of milk β-lactoglobulin and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Screening of Lactobacillus plantarum AHQ-14

[0022] 1. bacterial strain isolation method: prepare MRS medium: according to 20g / L glucose, 10g / L peptone, 5g / L yeast extract, 10g / L beef extract, 5g / L sodium acetate, 2g / L diamine hydrogen citrate, 2g / L Dipotassium Phosphate, 0.58g / L MgSO 4 ·7H 2 O, 0.25g / L MnSO 4 ·5H 2 O and 1mL / L Tween-80 were weighed raw materials, mixed uniformly, dissolved in deionized water, the obtained solution was sterilized at a temperature of 121 ° C for 15min to obtain the MRS medium; 15g / L agar powder was added to the medium , sterilized at a temperature of 121 °C for 15 min to obtain MRS agar medium.

[0023] Mix the collected Xinjiang traditional fermented yogurt samples with a vortex shaker, weigh 0.5g or 0.5ml of the sample into 4.5ml of sterile saline with a concentration of 0.85%, and dilute the sample ten-fold gradient (10 -4 , 10 -5 , 10 -6 ), then spread on MRS agar medium or M17 agar plate medium, cultured anaero...

Embodiment 2

[0025] Example 2: Morphological characteristics and bacterial species identification of Lactobacillus plantarum AHQ-14

[0026] 1. Colony morphological characteristics: Lactobacillus plantarum AHQ-14 of the present invention forms milky white colonies on MRS agar medium, opaque, round, neat edges, central bulge, about 1.3~1.6mm in diameter, see attached for details. figure 1 .

[0027] II. Bacterial morphology: Lactobacillus plantarum AHQ-14 of the present invention has the following morphological characteristics: the cells are long rod-shaped with blunt ends, 4.0-8.0 μm long and 0.9-1.1 μm wide, and are single or chain-shaped. For details, see attached figure 2 .

[0028] III. Molecular biological identification: using universal primers, the forward primer is 27f: 5′-AGAGTTTGATCCTGGCTCAG-3′; the reverse primer is 1492r: 5′-GGTTACCTTGTTACGACTT-3′, and the bacterial DNA is used as the amplification template for PCR amplification 16S rRNA gene region, the amplified product w...

Embodiment 3

[0030] Example 3: Bile salt hydrolase activity of Lactobacillus plantarum AHQ-14

[0031] The strains were supplemented with 0.2% (w / v) sodium thioglycolate, 0.3% (w / v) sodium taurodeoxycholate hydrate and 0.2% (w / v) CaCl 2 cultured on MRS agar plates. The sterilized MRS medium is added to the sterile petri dish, and a sterile filter paper sheet with a diameter of about 3-4 mm is evenly placed on the solidified medium. Adsorb 10 μL of activated bacterial solution, slowly add filter paper until the liquid no longer flows, and culture at 37 °C for 3 days under anaerobic conditions. The precipitation area around the colony shows the bile salt hydrolase activity of the strain.

[0032] MRS medium was used as a negative control and LGG was used as a positive control for qualitative determination of bile salt hydrolase activity. Figure 4 A large amount of deoxycholic acid precipitated and diffused into the medium around AHQ-14, which proved the existence of bile salt hydrolase a...

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Abstract

The invention discloses a strain of plant lactobacillus that reduces the antigenicity of milk β-lactoglobulin and its application, belonging to the technical field of biological applications. The strain was deposited at the Chinese Type Culture Collection Center of Wuhan University, Bayi Road, Wuchang District, Wuhan City on October 10, 2020, with the preservation number CCTCC NO: M2020544. Lactobacillus plantarum AHQ‑14 has bile salt hydrolyzing enzyme activity, high adhesion to intestinal cells, high survival rate in gastrointestinal fluid, probiotics, low production of biogenic amines, no hemolysis, and sensitivity to commonly used antibiotics. Lactobacillus plantarum AHQ-14 is applied to foods containing β-lactoglobulin, which can effectively degrade β-lactoglobulin in milk, thereby reducing the antigenicity of milk protein and reducing the occurrence of allergic reactions.

Description

technical field [0001] The invention belongs to the technical field of biological applications, relates to a Lactobacillus plantarum AHQ-14 with reduced beta-lactoglobulin sensitization, and also relates to the use of the Lactobacillus plantarum AHQ-14. Background technique [0002] Cow's milk proteins commonly cause allergic reactions in children, and about 2-3% of infants and young children have symptoms of cow's milk protein allergy. β-lactoglobulin in cow's milk is considered to be the main allergen causing milk allergy in children. There are many methods to reduce the sensitization of β-lactoglobulin, such as heating, glycosylation, enzymatic hydrolysis, etc. However, heat treatment may generate more epitopes and enhance allergy. Glycosylation will cause the loss of some nutrients. There are many types of enzymes, and the mode and degree of action of enzymes under different conditions are also different, resulting in different decomposition products and antigenicity. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23C9/123A23C19/032A23C19/06C12R1/25
CPCA23C9/1234A23C19/0323A23C19/062A23V2400/169
Inventor 姜淑娟孟令莹牟光庆朱雪梅妥彦峰钱方
Owner DALIAN POLYTECHNIC UNIVERSITY
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