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Bacillus thuringiensis DavIV and application thereof

A technology of Bacillus thuringiensis and Bacillus aureus, applied in the field of microorganisms

Inactive Publication Date: 2019-04-12
SICHUAN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The present invention obtains a strain of Bacillus thuringiensis (Bacillus thuringiensis) from the asparagus planting soil of Sichuan Normal University by using the plate dilution coating method and the streaking method on LB medium, and the test finds that the strain has strong protein decomposition ability , so far there is no relevant report

Method used

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  • Bacillus thuringiensis DavIV and application thereof
  • Bacillus thuringiensis DavIV and application thereof
  • Bacillus thuringiensis DavIV and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Sample Collection, Strain Isolation

[0018] The characteristics of the colonies were separated by dilution coating and plate streaking. Take out the strains from the refrigerator at 4°C, use an inoculation loop to pick up a small amount of strains and inoculate them on the beef extract peptone medium, and place the beef extract peptone medium in a constant temperature incubator (37°C) for 48 hours , then observe. The edge of the colony is notched, showing a round shape, and the surface is moist, making it difficult to pick.

Embodiment 2

[0019] The identification of embodiment 2 bacterial strains

[0020] The purified strain was subjected to 16S rRNA gene amplification and PCR amplification to obtain a specific band of about 1.5 kb, and a DNA sequence as shown in SEQ ID NO.1. Comparing the homology between the GenBank nucleic acid sequence library and the obtained sequence, it was found that the strain DavⅣ had the closest relationship with the genus Bacillus, and the homology with Bacillus Coagulans was the highest, reaching 99%. The bacterium was identified as Bacillus thuringiensis (Bacillus thuringiensis) according to 16S rDNA sequence homology and morphological analysis, so the strain DavIV obtained in the present invention was named Bacillus thuringiensis (Bacillus thuringiensis) DavIV. The colony grown on the TSA plate is off-white with wavy edges, and the growth temperature range is 15-45°C. The strain was puncture-inoculated into beef extract peptone semi-solid medium, and the strain was aerobic and ...

Embodiment 3

[0023] The enzyme activity experiment of embodiment 3 bacterial strains

[0024] Enzyme preparation

[0025] Pick a small amount of bacterial lawn and put it in 50mL beef extract peptone liquid medium, cultivate it on a shaker at 37°C and 180rpm for 24h, and take the supernatant to obtain the crude enzyme solution.

[0026] Determination of protease activity

[0027] Protease activity was determined by the Folin method. After 24 hours of culture, the bacteria were taken and centrifuged at a low speed, and the supernatant was taken to measure the protease activity. Take 1ml of the sample and add it to a centrifuge tube containing 1ml of 1% casein solution (phosphate buffer solution with a pH value of 6.8) that has been pre-warmed at 30°C, 40°C, 50°C, and 60°C. 2ml of trichloroacetic acid with a concentration of 0.4mol / L to terminate the reaction, let it stand for 30min at room temperature, take 1ml of the supernatant after centrifugation and put it in a test tube, add 5ml of...

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Abstract

The invention discloses a Bacillus thuringiensis DavIV and application thereof. The strain is deposited in China Center for Type Culture Collection on September 30, 2018, and the preservation number is CCTCC NO:M 2018626. The strain has nicked edges, a round shape and a relatively moist surface, and is difficult to pick. Tests prove that the Bacillus thuringiensis DavIV has good protein decomposition ability.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacillus thuringiensis Dav IV and an application thereof. Background technique [0002] Bacillus thuringiensis is an aerobic Gram-positive bacterium in the Bacillus cereus group (Bacillus cereus). ℃-40℃. Usually, endospores are formed under the external environmental conditions in which it senses adversity. These spores have a solid protective shell, which is a very hard structure that can be dormant and cannot be replicated. It can germinate and restore nutrition when the external environmental conditions are suitable. grow and divide. [0003] The present invention obtains a strain of Bacillus thuringiensis (Bacillus thuringiensis) from the asparagus planting soil of Sichuan Normal University by using the plate dilution coating method and the streaking method on LB medium, and the test finds that the strain has strong protein decomposition ability , so f...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00C12R1/07
CPCC12P21/00C12R2001/075C12N1/205
Inventor 马骏张宇冲何兵廖洋赵仕林
Owner SICHUAN NORMAL UNIVERSITY
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