A strain of Bacillus amyloliquefaciens with high fungal inhibitory activity and its application

A technology of amyloliqueficiating spores and inhibiting activity, which is applied to Bacillus amyloliquefaciens and its application in black pepper storage. It can solve the problems of drug-resistant bacterial antibiotic residues and food safety impacts, and achieve strong pH stability and thermal stability. Stability, the effect of reducing the content of aflatoxin

Active Publication Date: 2018-07-20
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the important means to prevent and control the pollution of yellow mold and its toxins is to use antibiotics and chemical agents, but this will lead to problems such as the emergence of drug-resistant bacteria and antibiotic residues, which will have a major impact on food safety. Therefore, it is urgent to develop new means to Prevention and Control of Mold and Its Toxin Pollution

Method used

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  • A strain of Bacillus amyloliquefaciens with high fungal inhibitory activity and its application
  • A strain of Bacillus amyloliquefaciens with high fungal inhibitory activity and its application
  • A strain of Bacillus amyloliquefaciens with high fungal inhibitory activity and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 The inhibitory effect of Bacillus amyloliquefaciens F1 on the growth of mold

[0019] Mold culture and spore suspension production: inoculate the mold stored on the PDA slant medium onto the PDA plate, and culture it in the incubator at 28°C for 4 days, then add a certain amount of sterile PBS buffer, lightly lighten it with the tip of a large sterile gun. Scrape the spores on the culture medium, then pass the suspension containing mold spores and hyphae through a blue-capped bottle with eight layers of sterilized gauze to filter out mold mycelium, vortex, and dilute 10 and 100 respectively by 10-fold dilution method , After 1000 times, count on a hemocytometer, and then adjust the counted fungal spore stock solution to 10 with sterile PBS buffer. 5 per mL, and stored in a refrigerator at 4°C for later use.

[0020] Determination of antibacterial rate: Thaw the isolated and preserved Bacillus amyloliquefaciens F1 at 4°C, absorb 100 μL of the bacterial solutio...

Embodiment 2

[0022] The bacteriostasis property of embodiment 2 Bacillus amyloliquefaciens F1 fermented liquid and ultrafiltrate thereof

[0023] Ultrafiltration membranes with membrane molecular weight cut-offs of 100ku, 30ku and 3ku were selected to perform ultrafiltration on the fermentation supernatant of strain F1, and the antibacterial activity of each part of the ultrafiltrate was as follows: image 3 As shown, the antibacterial activity of 100ku filtrate was slightly higher than that of the original supernatant; the antibacterial activity of 30ku filtrate and 3ku filtrate decreased by 65.31% and 80.61%, respectively. In summary, the molecular weight of antifungal substances produced by strain F1 ranged from 30ku to 100ku. In order to further confirm whether the substance is a protein substance, alkaline protease, trypsin and proteinase K were used respectively, and the ultrafiltrate (30ku ~ 100ku) of strain F1 was used as the substrate for enzymolysis to detect its antibacterial ac...

Embodiment 3

[0025] Example 3 Effect of Bacillus amyloliquefaciens F1 antibacterial protein on the growth of Aspergillus flavus hyphae

[0026] Aspergillus flavus spore suspension preparation method is the same as embodiment 1, and spore concentration is adjusted to 10 6individual / mL. Add PDA (potato dextrose liquid medium) 150mL to each Erlenmeyer flask, put it to room temperature after autoclaving, add 300 μ L of Aspergillus flavus spore suspension to each Erlenmeyer flask, the blank control group does not add Bacillus amyloliquefaciens F1 ultrafiltration antibacterial liquid, The other two groups were added with 10% and 20% volume ratio of ultrafiltration antibacterial solution respectively, each group was set up in three parallels, 150r / min, 30°C shake flask culture for 3 days, medium-speed qualitative filter paper was used to filter the culture solution, and distilled water was washed three times. Remove the viscous components of the medium, put the mycelium obtained by filtration in...

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Abstract

The invention relates to a strain of bacillus amyloliquefaciens with high fungal inhibitory activity and application thereof, belonging to the field of biotechnology application. The invention provides a new bacterial strain; a Bacillus amyloliquefaciens, the preserved strain name is Bacillus amyloliquefaciens F1, its preservation number is CGMCCNo.10942, the preservation date is June 1, 2015, and the preservation unit is China Microbial Strain Collection Committee General Microbiology Center (CGMCC). The Bacillus amyloliquefaciens F1 with the preservation number of CGMCC No. 10942 of the present invention can produce antifungal protein, and can effectively inhibit the growth and toxin production of fungi such as Aspergillus niger, Aspergillus white, Aspergillus parasitica and Aspergillus flavus. The antibacterial protein produced by the bacterial strain of the invention can resist the infection of the mold on the black pepper, weaken the growth ability of the mold hyphae, protect the active ingredients of the black pepper, and reduce the loss and deterioration of the quality of the black pepper. The bacterial strain and its metabolites of the invention can be developed into a residue-free, safe and efficient food protection agent, which prolongs the shelf life of food and improves food safety.

Description

technical field [0001] The invention belongs to the field of microorganism and biotechnology, and in particular relates to a bacillus amyloliquefaciens with high fungal inhibitory activity screened from the surface of commercially available black pepper and its application in black pepper storage. Background technique [0002] Filamentous fungi and yeasts are common groups of microorganisms responsible for the spoilage of foods such as fermented dairy products, cereals, fruits and vegetables, and grains. It is estimated that 5% to 10% of food loss worldwide is caused by fungi. In Western Europe, bread spoilage caused by mold costs £2 billion a year in economic losses. Penicillium and Aspergillus have been reported to be the main fungal microorganisms causing food spoilage. my country is a large agricultural country. Due to the widespread occurrence and serious damage of crop diseases and insect pests, more than 10 million tons of grain are lost every year due to diseases a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L3/3463C12R1/07
Inventor 方维明饶胜其尹永祺汪志君高璐
Owner YANGZHOU UNIV
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