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Culture medium for culturing living biological medicine and application of culture medium

A technology of living organisms and culture medium, applied in the field of microorganisms, can solve the problems of limited proliferation promotion of probiotics, achieve significant drug effects, prevent or treat non-alcoholic fatty liver disease, and promote growth and proliferation

Active Publication Date: 2022-07-29
JIANGSU WECARE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the improved probiotic culture medium proposed in the prior art is limited to the promotion of the growth and proliferation of probiotics
On the one hand, these improved media have a limited effect on promoting the proliferation of probiotics. On the other hand, the improvement of the corresponding functions of probiotics can only be achieved by increasing the number of viable bacteria. For a certain number of viable bacteria, it can be achieved by changing the culture conditions Further improving the function of probiotics has not been reported

Method used

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  • Culture medium for culturing living biological medicine and application of culture medium
  • Culture medium for culturing living biological medicine and application of culture medium
  • Culture medium for culturing living biological medicine and application of culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 - Preparation of culture medium

[0047] Formula 1: By mass percentage: peptone 1%, beef extract 1%, lactulose 0.6%, oligomannose 1.6%, raffinose 0.8%, yeast extract 0.5%, diammonium hydrogen citrate 0.2% %, K 2 HPO 4 ·3H 2 O 0.2%, MgSO 4 ·7H 2 O 0.058%, MnSO 4 0.019%, Tween-80 0.1%, L-cysteine ​​0.1%, riboflavin 0.1%, asparagine 0.1%, and the balance is water.

[0048] Formula 2: By mass percentage: peptone 0.6%, beef extract 1.5%, lactulose 0.4%, mannose oligosaccharide 2%, raffinose 0.6%, yeast extract 0.8%, diammonium hydrogen citrate 0.05% %, K 2 HPO 4 ·3H 2 O 0.5%, MgSO 4 ·7H 2 O 0.1%, MnSO 4 0.01%, Tween-80 0.3%, L-cysteine ​​0.03%, riboflavin 0.05%, asparagine 0.15%, and the balance is water.

[0049] Formula 3: By mass percentage: peptone 1.5%, beef extract 0.5%, lactulose 0.8%, oligomannose 1.2%, raffinose 1%, yeast extract 1%, diammonium hydrogen citrate 0.3% %, K 2 HPO 4 ·3H 2 O 0.05%, MgSO 4 ·7H 2 O 0.02%, MnSO 4 0.08%, Twee...

Embodiment 2

[0065] The present embodiment provides a culture of living biological medicine LBPs1 (combination of Lactobacillus reuteri LR08 and Lactobacillus paracasei LC86, weight ratio 3:1) for preventing or treating asthma, and the preparation method is as follows:

[0066] A. Activation: Take out Lactobacillus reuteri LR08 and Lactobacillus paracasei LC86 stored in a 40% glycerol tube in a -80°C refrigerator. After the glycerol tube is thawed, mix the samples at a weight ratio of 3:1. Pipet 2% of the sample in a sterile operating table and add it to 20 mL of modified MRS medium. After culturing at 37 °C for 18 h, take out and centrifuge (8000 g for 10 min) to obtain the separated bacterial slurry, which is then used in 9 mL. Resuspend in sterile physiological saline and shake evenly to obtain bacterial liquid.

[0067] B. Cultivation: The culture medium of formula 1 to formula 15 in Example 1 was used to cultivate the living biopharmaceutical LBPs1. Dispense the medium into centrifug...

Embodiment 3

[0092] This example provides a culture of living biological drug LBPs2 (combination of Lactobacillus acidophilus LA88 and Lactobacillus rhamnosus LRa05, weight ratio 1:1) for the prevention or treatment of non-alcoholic fatty liver disease.

[0093] A. Activation: Refer to Step A of Example 2.

[0094] B. Cultivation: Refer to step B of Example 2.

[0095] C Colony growth rate comparison: with reference to step C of Example 2, the results are shown in Table 4.

[0096] Table 4

[0097]

[0098] D Bacterial proliferation ability comparison: with reference to step D of Example 2, the results are shown in Table 5.

[0099] table 5

[0100]

[0101] It can be seen from the results in Table 4 and Table 5: on the premise that the total amount is consistent, the present invention combines lactulose, oligomannose and raffinose, which has a more significant effect than the formula lacking one component. Compared with the conventional medium (MRSC), the growth rate of the colo...

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Abstract

The invention relates to a culture medium for culturing a living biological medicine and application of the culture medium. The culture medium for culturing the living biological medicine is prepared from peptone, beef extract, lactulose, mannose oligomer, raffinose, yeast extract, diammonium hydrogen citrate, K2HPO4, MgSO4, MnSO4, Tween-80 and L-cysteine. The living biological medicine comprises any one or a combination of at least two of lactobacillus reuteri, lactobacillus paracasei, lactobacillus acidophilus, lactobacillus rhamnosus, bifidobacterium longum, pediococcus pentosaceus, lactobacillus crispatus, lactobacillus jensenii or lactobacillus gasseri. The culture medium has an unexpected promoting effect on growth, proliferation, metabolism and efficacy exertion of living biological medicines. The efficacy is such as prevention or treatment of gastrointestinal diseases, immune diseases, cancers, vaginitis and other diseases.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to a culture medium for culturing living biological medicines and its application, in particular to a culture medium for promoting the growth, proliferation and efficacy of living biological medicines and its application. Background technique [0002] Live biotherapeutic (LBPs) refers to live organisms (bacteria, etc.) that are suitable for the prevention, treatment or cure of a certain disease in humans. They are neither vaccines nor filterable viruses. Generally speaking, they will not be administered by injection. It is an emerging and disruptive new drug. For example, fecal microbiota transplantation (FMT) is a living biological drug that was researched and developed earlier, but its quality and safety are difficult to control, the fecal flora cannot be preserved for a long time, and industrialized and standardized production cannot be realized, and the FMT process and dono...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/38A61K35/747A61K35/745A61K35/744A61P31/04A61P31/10A61P37/08A61P11/00A61P1/16A61P35/00A61P15/02C12R1/225C12R1/23C12R1/01
CPCC12N1/20C12N1/38A61K35/747A61K35/745A61K35/744A61P31/04A61P31/10A61P37/08A61P11/00A61P1/16A61P35/00A61P15/02C12R2001/225C12R2001/23C12R2001/01A61K2300/00
Inventor 方曙光董瑶盖忠辉朱建国
Owner JIANGSU WECARE BIOTECHNOLOGY CO LTD
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