Marine bacillus and its application for lipopeptide production

A technology of marine bacillus and bacillus, which is applied in the biological field to achieve the effect of improving the aquaculture environment and promoting the aggregation and sedimentation of bacterial cells

Active Publication Date: 2020-07-17
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to reduce the problems of increased resistance of pathogenic bacteria caused by the abuse of traditional antibiotics and chemical drugs and the destruction of aquaculture food safety, environmentally friendly antibacterial active substances need to be further developed and widely used

Method used

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  • Marine bacillus and its application for lipopeptide production
  • Marine bacillus and its application for lipopeptide production
  • Marine bacillus and its application for lipopeptide production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The marine bacillus Bacillus sp.BS176 is isolated from the seamount region of the western Pacific Ocean. The bacterium can grow in 2216E medium. The 16S rDNA of Bacillus sp.BS176 is shown in SEQ ID NO.1.

[0033] The isolated marine Bacillus sp.BS176 was inoculated into 2216E liquid medium, and cultured overnight at 28°C with shaking. Take 5 mL of the bacterial liquid and transfer it to 500 mL of fermentation medium (peptone 20 g / L, yeast powder 10 g / L, glucose 5 g / L, seawater 1 L, pH 7.5), and culture with shaking at 28°C for 2 days.

[0034] After the fermentation, centrifuge at 8000 rpm for 10 minutes to collect the fermentation supernatant, adjust the pH of the supernatant to about 2.0 with 6N HCl solution, and then put it in a chromatographic cabinet at 4°C overnight. The overnight solution was centrifuged at 8000 rpm for 10 min at 4° C. to collect the precipitate. The obtained precipitate was washed three times with a pH 2.0 hydrochloric acid solution, and then t...

Embodiment 2

[0047] Example 2 Bacillus sp.BS176 inhibits the activity of Vibrio alginolyticus motility

[0048] Bacillus sp.BS176 and Vibrio alginolyticus were inoculated in 5 mL 2216E liquid medium respectively, and cultured overnight at 28°C with shaking. Dilute the bacterial solutions of these two bacteria with sterile 2216E liquid medium to OD 600 After the value is 0.2, take 1 μL of Vibrio alginolyticus V.alginolyticus bacteria solution and inoculate it in the center of 2216E solid medium, then take 1 μL of Bacillus sp.BS176 and inoculate it in the above-inoculated 2216E solid medium Next to Vibrio V. alginolyticus, the two bacteria were co-cultured at 28 °C. After the cultivation, it was found that on the medium inoculated with Bacillus sp.BS176, the movement of the colony of Vibrio alginolyticus was significantly inhibited, indicating that Bacillus sp.BS176 can inhibit the growth of Vibrio alginolyticus V. The exercise capacity of alginolyticus ( figure 2 A, wherein A-a, Vibrio ...

Embodiment 3

[0050] Example 3: The activity of Bacillus sp.BS176 lipopeptide molecules to promote the aggregation and sedimentation of Vibrio alginolyticus cells

[0051] Vibrio alginolyticus V.alginolyticus was inoculated in 5mL 2216E liquid medium and cultured overnight at 28°C with shaking, then inoculated into fresh 5mL 2216E liquid medium at a volume ratio of 1:100, and continued to culture at 28°C to OD 600Value 0.2-0.3. At this time, the lipopeptide solution CLPs with a concentration of 10 mg / mL was added to the culture solution of Vibrio alginolyticus V. alginolyticus at a dilution of 1:100 (volume ratio) at 28 ° C for 3 hours, while the control group bacterial solution 50 μL of DMSO was added to the medium. After the cultivation, the alginolyticus V.alginolyticus added the lipopeptide solution showed the phenomenon of thalli agglutination, and the agglutinated thalli quickly settled to the bottom of the test tube ( image 3 A).

[0052] The 96-well plate method was used to quan...

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Abstract

The invention belongs to the field of biotechnology, and relates to applications of marine bacillus (Bacillus sp. BS176) and lipopeptide molecules. The lipopeptide-producing marine bacterium is Bacillus sp.BS176, which is preserved in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing (Institute of Microbiology, Chinese Academy of Sciences), and the date of preservation As of December 2016, deposit number: CGMCC No.13515. The bacillus in the present invention is isolated from the seamount region of the western Pacific Ocean and belongs to marine microorganisms. The lipopeptide produced by the bacterium has the activity of inhibiting the motility of pathogenic bacteria in aquaculture, and has the effect of promoting bacterial agglutination and sedimentation for various pathogenic bacteria. The bacillus and the lipopeptide produced by it have potential application value in developing broad-spectrum antibacterial drugs and improving aquaculture environment.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to applications of marine bacillus (Bacillus sp. BS176) and lipopeptide molecules. Background technique [0002] Biosurfactants are a class of surface active substances derived from living organisms, especially microorganisms. According to the structural characteristics of biosurfactants, they can be divided into five categories: glycolipids, lipopeptides and lipoproteins, phospholipids and fatty acids, polymeric surfactants and particulate surfactants. Due to the characteristics of mild production conditions, low toxicity, biodegradability and environmental compatibility, biosurfactants have become excellent substitutes for chemical surfactants, and have potential application advantages in industrial production. They can be used in food, cosmetics, and pharmaceutical industries. , environmental protection and energy-saving technology [1]. [0003] Lipopeptide is an important class of b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C07K14/32C12P21/02A61K38/16A61P31/04C12R1/07
CPCC07K14/32A61K38/00C12N1/20A61P31/04C12R2001/07C12N1/205
Inventor 孙超岷张德超刘瑞修鹏远
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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