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Bacillus subtilis capable of producing broad spectrum and efficient antibacterial peptide and application of Bacillus subtilis

A technology of Bacillus subtilis and antimicrobial peptides, applied in the field of microorganisms, can solve the problems of less research on the application of antimicrobial peptides, and achieve the effects of wide application range, no loss of bacteriostatic activity, and broad antibacterial spectrum.

Inactive Publication Date: 2013-10-02
XUZHOU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, many natural antimicrobial peptides have been studied and discovered, but only Nisin and Natamycin are the most widely used bacteriocins in food, and the application research of other antimicrobial peptides is less

Method used

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  • Bacillus subtilis capable of producing broad spectrum and efficient antibacterial peptide and application of Bacillus subtilis
  • Bacillus subtilis capable of producing broad spectrum and efficient antibacterial peptide and application of Bacillus subtilis
  • Bacillus subtilis capable of producing broad spectrum and efficient antibacterial peptide and application of Bacillus subtilis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Screening of BRT39 strain

[0018] (1) Take 2g of soil sample from the vegetable garden with rich organic matter content, add it to a small triangular flask containing 50mL of cooled sterile water, shake at 200r / min for 20min, and then put it in a water bath at 80°C for 10min, continuously Shake the flask to mix the soil sample, let it stand for 5 minutes, draw 100 μL of the supernatant, and dilute it to 10 in sequence. -1 ~10 -9 concentration, choose 10 -3 , 10 -4 , 10 -5 , 10 -6 Concentration coated beef extract peptone plate, incubated at 37°C for 24h, and continued to be incubated at 30°C for 24h.

[0019] (2) According to the characteristics of the shape, size, surface structure, edge structure, texture, gloss, transparency, color and soluble pigment produced, the isolated single colony is picked out with an inoculation loop and moved to the screening culture On the base plate, numbered and incubated at 34°C for 48h.

[0020] The screening medium ...

Embodiment 2

[0027] Example 2: Strain identification

[0028] Physiological and biochemical identification of strains: carbon source utilization test, catalase test, hydrogen sulfide test, indole test, nitrate and nitrite reduction test, etc. ; Morphological identification: The strains to be identified were seeded on a solid plate of beef extract peptone medium, cultured at 37°C for 2 days, and the culture characteristics of the colony and the morphological characteristics of the bacteria were described and recorded. Physiological and biochemical identification characteristics are shown in Table 1. The results in the table are consistent with the species characteristics of Bacillus subtilis according to the "Berger's Bacterial Identification Manual". Molecular biological identification: The extraction of bacterial genomic DNA refers to the method of "Molecular Cloning Experiment Guide" (3rd Edition). Primers were designed based on the most conserved sequence in bacterial 16S rDNA. Prime...

Embodiment 3

[0032] Example 3: Determination of antibacterial activity of bacterial strain fermentation broth

[0033] (1) The fermentation broth of strain BRT39 was centrifuged at 10,000 r / min for 5 min, and the supernatant was filtered through a 0.22 μm microporous membrane and used for the determination of antibacterial activity.

[0034] (2) The indicator bacteria were cultivated in nutrient agar medium, and the antibacterial activity of the shake flask fermentation broth was directly determined by punching method. Nutrient agar medium plate coating cultured 100μL indicator bacteria suspension (1 × 10 8 cfu / ml), uniformly punch holes with a 6 mm diameter hole punch, add 100 μL of the prepared fermentation broth to each hole, and use sterile water as a control. Incubate at 37°C for 16h, and record the size of the inhibition zone. Select Escherichia coli, Staphylococcus aureaus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella enteritidis, Bacillus subtilis, etc. Bacteria were used...

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Abstract

The invention discloses Bacillus subtilis capable of producing broad spectrum and efficient antibacterial peptide and an application of the Bacillus subtilis and belongs to the technical field of microorganisms, wherein the Bacillus subtilis is Bacillus subtilis BRT39 concretely with the preservation number of CGMCC NO. 5702. Antibacterial peptide in metabolic products of a BRT39 bacterial strain has stronger anti-bacterial effect, a fermentation liquor can inhibit gram positive bacteria and also has stronger inhibiting effect on multiple gram negative bacteria and fungi, and the antibacterial peptide in the fermentation liquor has strong acid-base stability, good thermal stability, strong light stability and high organic solvent tolerance. The antibacterial peptide produced by the BRT39 bacterial strain belongs to a natural antibacterial agent, has no side or toxic effect, can be used as a food preservative, a feed additive, an agricultural biological control agent and an animal disease prevention agent and has the advantages of no toxicity, no residue, no drug resistance and high safety.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a Bacillus subtilis strain producing broad-spectrum and efficient antibacterial peptides and applications thereof. Background technique [0002] Under suitable environmental conditions, microorganisms in food multiply, resulting in food spoilage. One of the key issues in food processing is preservation and preservation. One of the important measures to extend the preservation period and make the food circulate and have commercial value is to add an appropriate amount of food preservatives and preservatives in the food processing process. Chemical preservatives such as benzoic acid, sorbic acid and its salts, parabens, etc. are the most widely used food preservatives, which can inhibit or kill microorganisms, prevent and delay food spoilage. Different from industrial preservatives, the safety and hygienic conditions of food preservatives are increasingly concerned, and co...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23L3/3463C12R1/125
Inventor 高兆建樊陈张抗震鞠民友杜永凯孙会刚王东星
Owner XUZHOU UNIV OF TECH
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