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

A Bacillus subtilis, antimicrobial peptide technology, applied in the field of microorganisms, can solve the problem of less application of antimicrobial peptides and other problems, and achieve the effects of wide application range, broad antibacterial spectrum and short production cycle

Inactive Publication Date: 2014-11-05
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] Embodiment 1: Screening of BRT39 bacterial strain

[0018] (1) Take 2g soil sample from the vegetable garden rich in organic matter, add it to a small triangular flask containing 50mL of cooled sterile water, shake the table at 200r / min, shake for 20min, then place it in a water bath at 80°C for 10min, and keep Shake the Erlenmeyer flask to mix the soil sample, let it stand for 5 minutes, absorb 100 μL of the supernatant, and gradually dilute to 10 -1 ~10 -9 Concentration, select 10 -3 , 10 -4 , 10 -5 , 10 -6 Concentration coated beef extract peptone plate, cultivated at 37°C for 24h, and continued to cultivate 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 of the microbial colony, use the inoculation loop to pick out the isolated single colony and move it to the screening culture Base plate, and numbered, 34 ℃ for 48h.

[0020] The scree...

Embodiment 2

[0027] Embodiment 2: bacterial strain identification

[0028] Physiological and biochemical identification of strains: Carry out carbon source utilization test, catalase test, hydrogen sulfide test, indole test, nitrate and nitrite reduction test and other tests according to the method of "Bergey's Bacteria Identification Manual" (Ninth Edition) Morphological identification: spot the strains to be identified on solid plates of beef extract peptone medium, culture at 37°C for 2 days, describe and record the characteristics of colony culture and morphological characteristics of bacteria. The physiological and biochemical identification characteristics are shown in Table 1. The results in the table are consistent with the characteristics of Bacillus subtilis species compared with "Bergey's Bacteria Identification Manual". Molecular biological identification: The extraction of bacterial genomic DNA refers to the method of "Molecular Cloning Experiment Guide" (3rd edition). Prime...

Embodiment 3

[0032] Embodiment 3: Determination of antibacterial spectrum activity of bacterial strain fermented liquid

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

[0034] (2) Use the nutrient agar medium to cultivate the indicator bacteria, and use the punching method to directly measure the antibacterial activity of the shake flask fermentation broth. Nutrient agar medium plates were coated with 100 μL indicator bacteria suspension (1×10 8 cfu / ml), uniformly punch holes with a hole puncher with a diameter of 6 mm, add 100 μL of the prepared fermentation broth to each hole, and use sterile water as a control. Incubate at 37°C for 16 hours, and record the size of the inhibition zone. Escherichia coli, Staphylococcus aureaus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella enteritidis, Bacillus subtilis, etc. Ba...

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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 microbes, in particular to a bacillus subtilis strain producing broad-spectrum high-efficiency antibacterial peptides and an application thereof. Background technique [0002] In the case of suitable environmental conditions, microorganisms in food multiply in large numbers, resulting in food spoilage. One of the key issues in food processing is anti-corrosion and fresh-keeping. And in order to reach prolonging the storage period, one of the important measures to make food circulate and have commodity value is to add an appropriate amount of food antiseptic and preservative in the food processing process. Chemical preservatives such as benzoic acid, sorbic acid and its salts, and parabens 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 sanitation conditions of food preservative...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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