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Separation and purification method of bacillus amyloliquefaciens antimicrobial proteins

A technology of amylolytic spores and antibacterial proteins, which is applied in peptide preparation methods, chemical instruments and methods, and organic chemistry. The effect of bacterial activity

Inactive Publication Date: 2011-08-17
INST OF MICROBIOLOGY HEILONGJIANG ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention aims to solve the problem that the existing protein separation and purification method cannot obtain high-purity Bacillus amyloliquefaciens TF28 antibacterial protein, many impurity proteins and low yield of antibacterial protein, and provides a separation and purification of Bacillus amyloliquefaciens antibacterial protein method

Method used

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  • Separation and purification method of bacillus amyloliquefaciens antimicrobial proteins

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specific Embodiment approach 1

[0016] Specific embodiment one: In this embodiment, the Bacillus amyloliquefaciens antibacterial protein is separated and purified according to the following steps:

[0017] 1. Take the supernatant of the fermentation broth of Bacillus amyloliquefaciens TF28, add solid ammonium sulfate to the supernatant until the concentration of ammonium sulfate is 20% saturation, then let it stand at 4°C for 2 hours, and then at a speed of 6000r / min Centrifuge for 30 minutes, collect the centrifuged supernatant, add solid ammonium sulfate to the centrifuged supernatant until the concentration of ammonium sulfate is 40% saturation, collect the precipitate, and dissolve the precipitate in phosphoric acid with a concentration of 0.02mol / L and a pH value of 6.8 Dialyze in the buffer solution and concentrate to obtain the crude extract of antibacterial protein;

[0018] 2. Use at room temperature The liquid chromatography system is used for Q Sepharose Fast Flow anion exchange chromatography. ...

specific Embodiment approach 2

[0026] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is: in step 2, the sample is manually loaded, and the sample volume is 100 μL. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0027] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that in step 3, the sample is loaded manually, and the sample volume is 100 μL. Other steps and parameters are the same as those in Embodiment 1 or 2.

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Abstract

The invention discloses a separation and purification method of bacillus amyloliquefaciens antimicrobial proteins, relating to a separation and purification method of bacillus antimicrobial proteins. The method solves the problems that the existing separation and purification method of proteins can not acquire bacillus amyloliquefaciens TF28 antimicrobial proteins with high purity, generates various impurity proteins and is low in the yield of antimicrobial proteins. The method includes the following steps: I. obtaining the crude extract of antimicrobial proteins; II. obtaining protein concentrate by anion exchange chromatography; and III. performing gel filtration chromatography through a molecular sieve to obtain bacillus amyloliquefaciens antimicrobial proteins. The separation and purification method is used for the separation and purification of bacillus amyloliquefaciens antimicrobial proteins.

Description

technical field [0001] The invention relates to a separation and purification method of bacillus antibacterial protein. Background technique [0002] Antibacterial proteins show strong antibacterial activity. In addition, antibacterial proteins can form ion channels on the bacterial plasma membrane, destroy the membrane potential, cause intracellular material leakage and kill bacteria, so it is not easy to produce drug resistance and cross resistance. Become the main research direction of new biopesticides. [0003] Although many bio-control microorganisms can secrete antibacterial proteins, and the separation and purification methods of proteins have been developed relatively well, but due to different strains, the properties of antibacterial proteins are very different, and the purification methods are also different. [0004] Bacillus amyloliquefaciens TF28 (Bacillus amyloliquefaciens TF28) belongs to the genus Bacillus, its preservation number is CGMCC No.4038, and its ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/36C07K1/30C07K1/18C07K1/16
Inventor 孟利强张淑梅赵晓宇李晶王玉霞张云湖
Owner INST OF MICROBIOLOGY HEILONGJIANG ACADEMY OF SCI
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