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Optimized human-body symbiotic lactobacillus peptidoglycan, and extraction method and application thereof

The technology of a lactobacillus and an extraction method is applied in the application field of vaccine adjuvants, which can solve the problems of poor biological activity of the final product and long extraction period, and achieve the effects of shortening the period and improving the immune enhancement effect.

Inactive Publication Date: 2015-04-22
CHONGQING ACADEMY OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide an optimized method for extracting the peptidoglycan of human symbiotic Lactobacillus, which solves the problems of too long extraction period and poor biological activity of the final product in the prior art

Method used

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  • Optimized human-body symbiotic lactobacillus peptidoglycan, and extraction method and application thereof
  • Optimized human-body symbiotic lactobacillus peptidoglycan, and extraction method and application thereof
  • Optimized human-body symbiotic lactobacillus peptidoglycan, and extraction method and application thereof

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Embodiment 1

[0054] An optimized human symbiotic Lactobacillus peptidoglycan extraction method is specifically implemented according to the following steps:

[0055] Step 1. Cultivation and amplification of human commensal Lactobacillus. The Lactobacillus was streaked and cultured on the MRS plate for 36 hours, and monoclonal colonies were picked, inoculated in 5 mL of MRS, and cultured at 37°C for 24-36 hours. Take the cultured bacterial liquid and inoculate it into 37.5mL MRS medium at a ratio of 1:100. According to the growth curve of the strain, culture it at 37°C for 36 hours to make it grow to the mid-logarithmic growth phase to obtain the bacterial sludge.

[0056] Step 2, separation and purification of peptidoglycan,

[0057] Collect the bacteria sludge cleaned by centrifugation, add trichloroacetic acid (TCA) with a mass concentration of 10% according to the solid-to-liquid ratio of 1:10, and shake well; shake it in a water bath at 60°C for 8 hours, cool it immediately, and centri...

Embodiment 2

[0059] An optimized human symbiotic Lactobacillus peptidoglycan extraction method is specifically implemented according to the following steps:

[0060] Step 1. Cultivation and amplification of human symbiotic Lactobacillus. Take human symbiotic Lactobacillus and culture it on the MRS plate for 32 hours, pick a single clone colony, inoculate it in 5 mL of MRS, and culture it at 37°C for 24 hours; take the cultured Inoculate 37.5mL MRS culture medium according to the volume ratio of 1:120, culture at 37°C for 32 hours to make it grow to the mid-logarithmic growth phase, and obtain the bacteria slime;

[0061] Step 2, the separation and purification of peptidoglycan, collect the bacteria sludge (wet weight) cleaned by centrifugation (8000g, 15min), add 10% trichloroacetic acid (TCA ) shake well; 55 ℃ water bath shaking treatment for 10h, cooling immediately, centrifugation (8000g, 15min); dehydrate the precipitate with methanol to remove TCA; degrease the residue with acetone; ...

Embodiment 3

[0063] An optimized human symbiotic Lactobacillus peptidoglycan extraction method is specifically implemented according to the following steps:

[0064] Step 1. Cultivation and amplification of human symbiotic Lactobacillus, take human symbiotic Lactobacillus and culture it on the MRS plate for 40 hours, pick a monoclonal colony, inoculate it in 5 mL of MRS, and culture it at 37°C for 36 hours; take the cultured Inoculate 37.5mL MRS culture medium according to the volume ratio of 1:80, culture at 37°C for 40 hours to make it grow to the mid-logarithmic growth phase, and obtain the bacteria slime;

[0065] Step 2, the separation and purification of peptidoglycan, collect the bacteria sludge (wet weight) cleaned by centrifugation (8000g, 15min), add mass concentration 10% trichloroacetic acid (TCA ) shake well; 65 ℃ water bath shaking treatment for 6h, cooling immediately, centrifugation (8000g, 15min); precipitate with methanol dehydration to remove TCA; residue degrease with a...

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Abstract

The invention discloses an optimized extraction method for human-body symbiotic lactobacillus peptidoglycan. The method comprises culture and amplification of human-body symbiotic lactobacillus, and separation and purification of peptidoglycan. The invention also discloses the human-body symbiotic lactobacillus peptidoglycan extracted by employing the optimized extraction method for human-body symbiotic lactobacillus peptidoglycan. The invention also discloses application of human-body symbiotic lactobacillus peptidoglycan extracted by employing the optimized human-body symbiotic lactobacillus peptidoglycan extraction method as a vaccine adjuvant of a mucosal immune system. The separation purification period of peptidoglycan is shortened, and the method is applicable to industrialized preparation of peptidoglycan. Peptidoglycan separated and purified by the method is verified to has good immunization enhancing effect, and is especially applicable as the vaccine adjuvant of the mucosal immune system.

Description

technical field [0001] The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to an optimized method for extracting the peptidoglycan of Lactobacillus symbiotica. In use as a vaccine adjuvant for the mucosal immune system. Background technique [0002] The mucosal immune system refers to the lymphatic tissue widely distributed in the submucosa of the respiratory tract, gastrointestinal tract, genitourinary tract, and some exocrine glands. A line of immune defense. The development of vaccines against the mucosal immune system has become an inevitable trend of development. In recent years, with the in-depth excavation of the physiological functions of human symbiotic bacteria, the important functions of the bacteria and their components in the mucosal immune system have become a research hotspot. Especially the peptidoglycan of human symbiotic Lactobacillus has been proved to have the development potential of AIDS vaccine adju...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04A61K39/39C12R1/225
Inventor 张华堂王超宋杰刘晓亮
Owner CHONGQING ACADEMY OF SCI & TECH
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