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A preparation method for inhibiting legionella active substance in bacillus hz16

A technology of Bacillus and Legionella, which is applied to the preparation of biologically active components by fermentation, the application of new drug development, and the field of activity of Bacillus strains, which can solve the problems of difficult treatment and killing of other diseases

Active Publication Date: 2020-11-13
ZHEJIANG UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of broad-spectrum antibiotics is that they may kill other beneficial or harmless microorganisms while acting on one kind of microorganism, and when drug resistance is formed, general drug resistance will be generated, which will bring difficulties to the treatment of other diseases in the future

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The preparation method of the active substance inhibiting Legionella in Bacillus HZ16 is prepared according to the following steps:

[0029] Step 1: Inoculate the purified Bacillus HZ16 on the LB solid medium with pH 5 under aseptic conditions, and cultivate for 1 day at 28 degrees Celsius to obtain white solid colonies;

[0030] Step 2. Inoculate the solid strains into a 300 ml Erlenmeyer flask containing 150 ml of LB medium with pH 5;

[0031] Step 3. Place the inoculated shake flask in a biological incubator, and cultivate 1 day-based fermentation at 28 degrees Celsius and 180 r / min to obtain fermentation broth;

[0032] Step 4. Use freeze-drying technology to remove water and compare the obtained solid dry weight;

[0033] Step 5. Extract with an organic solvent ethyl acetate to obtain a crude extract, which is concentrated for later use;

[0034] Step 6. Verify that the biologically active peptide obtained in Example 1 has highly selective inhibition of the biological activit...

Embodiment 2

[0037] Step 1. Inoculate purified Bacillus HZ16 on LB medium with pH 8 under aseptic conditions, and cultivate for 1 day at 28 degrees Celsius to obtain white solid colonies;

[0038] Step 2: Inoculate the solid hyphae into 150 ml of M with pH 8 2 50% SW medium in a 300 ml Erlenmeyer flask;

[0039] Step 3: Place the inoculated shake flask in a biological incubator, and cultivate 4 days-based fermentation at 28 degrees Celsius and 200 r / min to obtain fermentation broth;

[0040] Step 4. Use freeze-drying technology to remove water and compare the obtained solid dry weight;

[0041] Step 5. Extract with an organic solvent ethyl acetate to obtain a crude extract, which is concentrated for later use;

[0042] Step 6. Dissolve the crude extract with DMSO to make a solution of 50 μg / mL, and perform the same test of inhibiting Legionella activity as in Step 6 of Example 1, and the 48-hour inhibition zone size is 22 mm.

Embodiment 3

[0044] Step 1. Inoculate the purified Bacillus HZ16 on the Gaussian medium with a pH of 8 under aseptic conditions, and cultivate for 4 days at 28 degrees Celsius to obtain white solid colonies;

[0045] Step 2: Inoculate the solid hyphae into a 300 ml Erlenmeyer flask containing 150 ml of Gaussian medium with a pH of 8;

[0046] Step 3: Place the inoculated shake flask in a biological incubator, and cultivate 4 days-based fermentation at 28 degrees Celsius and 200 r / min to obtain fermentation broth;

[0047] Step 4. Use freeze-drying technology to remove water and compare the obtained solid dry weight;

[0048] Step 5. Extract with organic solvent ethyl acetate to obtain a crude extract, which is concentrated for later use;

[0049] Step 6. Dissolve the crude extract in DMSO to make a solution of 50 μg / mL, and perform the same test of inhibiting Legionella activity as in Step 6 of Example 1, and the 48-hour inhibition circle size is 18 mm.

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Abstract

The invention discloses a bacillus capable of producing active substances inhibiting legionella and a method for preparing active substances inhibiting legionella by using the bacillus. The present invention proves for the first time the activity of the metabolites of the strain to inhibit legionella, and uses the activity as an evaluation standard to study the fermentation conditions for producing stronger activity of the strain. The aim is to provide a new way to use Bacillus to produce active substances that inhibit Legionella, and to provide a scientific basis for further searching and developing effective drugs for the treatment of pneumonia.

Description

Technical field [0001] The invention relates to the field of development and utilization of microbial active substances, in particular to the activity of a Bacillus strain, a process method for preparing biologically active components by fermentation, and the application of the products in the development of new drugs. Background technique [0002] A strain of Bacillus HZ16 showed unique high selective inhibitory activity against Legionella. Different from the general characteristics of Bacillus that produce broad-spectrum antibiotics, the inhibitory activity of this strain against Gram-positive and negative bacteria other than Legionella is too weak or not detected. The disadvantage of broad-spectrum antibiotics is that they may kill other beneficial or harmless microorganisms while acting on one kind of microorganisms, and produce general drug resistance when drug resistance is formed, which will bring difficulties to the treatment of other diseases in the future. [0003] The g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/742A61P11/00A61P31/04C12R1/07
CPCA61K2035/115C12N1/205C12R2001/07Y02A50/30
Inventor 王宏鹏李音于林凯吴元锋黄俊杨瑞芹刘士旺毛建卫
Owner ZHEJIANG UNIVERSITY OF SCIENCE AND TECHNOLOGY
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