Bacillus velezensis and application thereof

A technology of Bacillus Velez and Bacillus, applied in the direction of bacteria, antibacterial drugs, medical preparations containing active ingredients, etc., can solve problems such as economic loss, decline in soil fertility, and impact on agricultural, forestry and animal husbandry production, and achieve improvement Yield, effectiveness of optimized fermentation methods

Active Publication Date: 2022-01-28
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, it will also lead to the decline of soil fertility, cause animal poisoning, affect the production of

Method used

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  • Bacillus velezensis and application thereof
  • Bacillus velezensis and application thereof
  • Bacillus velezensis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] This example provides a separation process of a Belice Bacillus EA73 strain, which is as follows:

[0033] (1) After taking sterile water, rinse the purple stem, stem, leaves, and flower tissue, use a sterile filter paper to dry moisture, first soak for 3 minutes with 3% NaClo, then use 0.1% HGCl 2 Soak 3 min, followed by soaking of 75% ethanol for 3 min, after flushing 5 times, the sterile filter paper was suiled.

[0034] (2) Take the last cleaning of the sterile water 0.2 ml, coated on the PDA medium, 25 ° C culture, and test whether the surface disinfection is thorough.

[0035] (3) The surface-sterilized plant samples were cut into 1 cm × 1 cm size fragment with a sterile blade, and inoculated to the LB agar medium. In the incubator at 30 ° C, it was observed on a day. After the edge of its incision was long, the bacterial bacterial plasma around the tissue block was selected to LB agar medium, and the four zone scribe Refute purification was repeated until the pure st...

Embodiment 2

[0043] In this example, morphology and molecular biological identification were performed on a strain EA 73 obtained in Example 1. The specific process is as follows:

[0044] 1) Morphological identification: The strongest strain in which the screening out of the antibacterial activity is inoculated on the LB plate, and after 37 ° C for 1D, the colonies are observed; and the Gram dye is observed under the microscope. Its bacterial shape. The strain EA 73 is cultured on the LB plate, which can be observed that the milk white, circular or elliptical, opaque, medium size, surface rough, edge neat, smooth mucosa; dyed mirror detection separation bacilli is Gram-positive Bacillus ,like figure 2 Indicated.

[0045] 2) Identification of molecular biology: Amplified the 16SRDNA sequence of the strain, the primer used:

[0046] Primers 27F: 5'-agagttgatcctggctcAg-3 ';

[0047] Primers 1492R: 5'-gcttaccttgttacgactt-3 ';

[0048] PCR amplification system (25 μL): Template DNA 2 μL, 2 × Taq P...

Embodiment 3

[0052] In this embodiment, the fermentation method of reinforcing the strain EA 73 in Example 1, specifically comprises the step of adding the Belce Bacillus to a fermentation medium, the fermentation medium, with yeast The carbon source is a nitrogen source in a nitrogen source, and 48 to 52 h are fermented at 27 to 28 ° C in NaCl. The optimum parameter of the fermentation process is controlled by: 2% induction amount, 2% inoculation, initial pH 7.95, fermentation time of 51.04 h, fermentation temperature 27.97 ° C, and rotational speed of 180 r / min.

[0053] The optimal conditions optimization process of the above fermentation method are as follows:

[0054] First, the medium composition is optimized

[0055] 1) Screening of optimal carbon sources and concentrations

[0056] In terms of the diameter of the bacterius ribs of Golden Staphylococcus, the best carbon source, nitrogen source and inorganic salts are selected from the single factor test, and their optimum concentratio...

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Abstract

The invention provides bacillus velezensis and application thereof. The preservation number of the bacillus velezensis is CCTCC NO: M20211139, the bacillus velezensis is submitted and preserved in the China Center for Type Culture Collection on September 6, 2021, and the bacillus velezensis is classified and named as Ea 73 (Bacillus velezensis Ea 73). The bacillus velezensis is separated from Eupatorium adenophorum endophyte, has strong antibacterial activity and is stable in heredity. The bacillus velezensis is further added into a fermentation culture medium, the fermentation culture medium takes yeast extract powder as a carbon source, peptone as a nitrogen source and NaCl as inorganic salt, fermentation is performed at 27-28 DEG C for 48-52 hours, and the bacteriostatic activity of the fermented strain on staphylococcus aureus is further enhanced. Therefore, the strain and the enhanced strain are expected to be developed into natural antibiotic products.

Description

Technical field [0001] The present invention belongs to the extraction and development technology of the plant endogenous bacteria, and is specifically related to a Belice Bacillus and its application thereof. Background technique [0002] In recent years, due to the abuse of antibiotic drugs, it has caused a pathogenic bacteria with drug resistance to seriously threatening the health of animals and humans. Therefore, finding new effective natural antibiotics is one of the important means of ensuring human health. Natural antibiotics mainly have natural animal and microbiological drugs, the finite and limitations of natural animal plant resources limit antibiotics, and microorganisms become an ideal material for antibiotic sources. [0003] Studies have shown that plants have generally existed in various plants, which is rich in cultivation, which cultures can produce many new secondary metabolites in the host or ex vivo, and have biological activity, such as antimicrobialism. Tu...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/742A61P31/04C12R1/07
CPCC12N1/20A61K35/742A61P31/04Y02A50/30
Inventor 胡延春谢雷文娟
Owner SICHUAN AGRI UNIV
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