Lactobacillus paracasei capable of efficiently expressing adenosine deaminase to relieve colitis

A Lactobacillus, paracheese technology, applied in the field of microorganisms, can solve the problems of tumor infection, fatigue, life-threatening and other problems, and achieve the effect of reducing the level of IL-1β

Pending Publication Date: 2022-08-05
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, long-term use can produce a variety of serious side effects, such as diarrhea, headache, fever, and fatigue, and even serious and potentially life-threatening infections and tumors

Method used

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  • Lactobacillus paracasei capable of efficiently expressing adenosine deaminase to relieve colitis
  • Lactobacillus paracasei capable of efficiently expressing adenosine deaminase to relieve colitis
  • Lactobacillus paracasei capable of efficiently expressing adenosine deaminase to relieve colitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1: Screening and identification of Lactobacillus paracasei CCFM1223

[0082] (1) strain screening

[0083] Taking feces from newborn healthy infants as samples, the samples were pretreated and stored in a -80°C refrigerator in 30% glycerol. After taking out and thawing, weigh 0.5g of the sample and add it to 4.5mL of 0.85% sterile saline for gradient. Dilute, select appropriate gradient dilution solution and spread it on MRS solid medium, culture at 37°C for 48h, pick typical colonies on MRS plate for streak purification, pick a single colony and transfer it to liquid MRS liquid culture based on 37°C culture 18h, 30% glycerol storage to obtain the strain.

[0084] (2) Identification of strains

[0085] The whole genome DNA of the extracted strain is used for the amplification of 16S rDNA, and the amplified DNA fragments are collected and sequenced (by Suzhou Anshengda Technology Co., Ltd., whose 16S rDNA sequence is shown in SEQ ID NO. , the results showed th...

Embodiment 2

[0089] Example 2: mRNA expression of Lactobacillus paracasei CCFM1223 adenosine deaminase

[0090] (1) Activation of strains:

[0091] The bacterial liquids of Lactobacillus paracasei CCFM1223 and Lactobacillus paracasei CCFM8631 were dipped from glycerol tubes and streaked on MRS solid medium, and cultured in a constant temperature incubator at 37 °C for 48 hours to obtain single colonies; single colonies were picked and inoculated in MRS In the liquid medium, the activated bacteria were obtained by culturing in a constant temperature incubator at 37° C. for 24 hours for activation cultivation, and repeating this operation 3 times.

[0092] (2) Inoculate the bacterial liquid obtained in step (1) into the MRS liquid medium at an inoculation amount of 2% (v / v), cultivate it in a constant temperature incubator at 37° C. for 24 hours to obtain a fermentation liquid, and place the obtained fermentation liquid at 12000 rpm After centrifuging for 10 min, collect the bacterial slurr...

Embodiment 3

[0098] Example 3: Effect of Lactobacillus paracasei CCFM1223 on the body weight of mice

[0099] Thirty-two SPF 7-week-old male C57BL / 6J mice were housed in an animal room with standard temperature and humidity, and the circadian standard of the animal room was strictly controlled, and sufficient standard mouse chow and drinking water were given.

[0100] After 7 days of adaptive feeding, they were randomly divided into 4 groups with 8 animals in each group. The 4 groups were: blank group, model group, positive control group and CCFM1223 group.

[0101] The 14-day intervention experiment procedure was as follows:

[0102] The mice in the blank group were given 0.2 mL of skim milk (concentration of 12.5 g / mL); they drank standard drinking water throughout the experiment;

[0103] Mice in the model group were given 0.2 mL of skim milk (concentration of 12.5 g / mL); standard drinking water was drank 7 days before the experiment, and dextran sodium sulfate (DSS) was added to the d...

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Abstract

The invention discloses lactobacillus paracasei capable of efficiently expressing adenosine deaminase to relieve colitis, and belongs to the technical field of microorganisms. The invention provides a strain of lactobacillus paracasei CCFM1223, which is preserved in the Guangdong Microbial Culture Collection Center on March 6, 2022, and the preservation number of the lactobacillus paracasei CCFM1223 is GDMCC No: 62271. The lactobacillus paracasei CCFM1223 has the advantages that the lactobacillus paracasei CCFM1223 is preserved in the Guangdong Microbial Culture Collection Center on March 6, 2022; the lactobacillus paracasei CCFM1223 provided by the invention can be used for improving loose and soft excrement, hematochezia and weight of mice with colitis; the increase of the proinflammatory factor level in the colon tissue is inhibited; the activity of antioxidant enzymes in colon tissues is increased; the content of malondialdehyde in colon tissues is inhibited; the activity of myeloperoxidase in colon tissue is inhibited; the lactobacillus paracasei CCFM1223 can increase the transcriptional level of a closely connected protein gene in a colon tissue, so that the lactobacillus paracasei CCFM1223 has a huge application prospect in preparation of a product for preventing and/or treating colitis.

Description

technical field [0001] The invention relates to a strain of Lactobacillus paracasei with high expression of adenosine deaminase for relieving colitis, and belongs to the technical field of microorganisms. Background technique [0002] Inflammatory bowel disease is characterized by noninfectious chronic inflammation of the gastrointestinal tract, mainly including Crohn's disease (which can affect any part of the gastrointestinal tract from the mouth to the anus), ulcerative colitis (limited to the colonic mucosa), and Definite colitis, of which ulcerative colitis is more common. Since the 1950s, the increasing incidence of ulcerative colitis and the decreasing age of onset have seriously affected the quality of life of patients and their families. The survey found that the highest incidence of ulcerative colitis is about 3 / 10,000, and the prevalence rate is as high as 5 / 1,000, causing a great burden on social medical costs. In recent years, the growth rate of ulcerative col...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/78A61K35/744A61P1/00A61P1/04A61P29/00A23L33/135A23L19/00A23L29/00A23L11/50A23C9/123C12R1/01
CPCC12N1/20C12N9/78C12Y305/04004A61K35/744A61P1/00A61P29/00A61P1/04A23L33/135A23L19/00A23L29/065A23L11/50A23C9/1234A23V2002/00A23V2400/165A23V2200/3204A23V2200/30
Inventor 唐鑫郭伟灵毛丙永崔树茂张秋香赵建新张灏陈卫
Owner JIANGNAN UNIV
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