Preparation method of bacillus subtilis HJDA32 and bacteriocin generated by bacillus subtilis HJDA32

A Bacillus subtilis, bacteriocin-producing technology, applied in the field of microorganisms, can solve problems such as food poisoning, spoilage, and adverse effects on the kidneys

Active Publication Date: 2013-04-03
SHANXI AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, one of the main methods to solve food safety is to add chemical preservatives, but long-term studies have found that excessive consumption of preservatives has adverse effects on blood pressure, heart, kidneys, etc., and even some chemical preservatives are carcinogenic, teratogenic and easily cause food poisoning
Thermal sterilization is also one of the important methods of killing microorganisms in food processing, but for some low-temperature products, such as low-temperature meat products, if the heating temperature is too high, the color and flavor will change significantly, and the nutritional value will also be greatly reduced
Other physical methods: such as ultraviolet disinfection, filter sterilization, cobalt 60 irradiation, etc., have no toxic effect on the human body, but when the package is opened, there is a possibility of re-contamination and corruption

Method used

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  • Preparation method of bacillus subtilis HJDA32 and bacteriocin generated by bacillus subtilis HJDA32
  • Preparation method of bacillus subtilis HJDA32 and bacteriocin generated by bacillus subtilis HJDA32
  • Preparation method of bacillus subtilis HJDA32 and bacteriocin generated by bacillus subtilis HJDA32

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Embodiment 1

[0033] A strain of Bacillus subtilis ( Bacillus subtilis ) HJD A32, which was deposited in the General Microorganism Center of China Committee for Culture Collection of Microorganisms (CGMCC for short) on September 25, 2012, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, zip code 100101), classified as Bacillus subtilis ( Bacillus subtilis ), the deposit number is: CGMCC No.6624.

[0034] The Bacillus subtilis ( Bacillus subtilis ) HJD A32 is isolated from the fermentation raw material of Shanxi mature vinegar. Cultured on beef extract peptone basal medium at 37°C for 48 hours, it is rod-shaped, arranged in single, pair or chain (see figure 1 ), the size is 0.6~0.85μm×1.5~3.5μm, with capsule (see figure 2 ), the motility is weak; the young bacteria are cultured for 2-18h, and Gram staining is negative; for 19-23h, Gram staining is negative and positive; for 24h, Gram staining is positive; Ro...

Embodiment 2

[0036] Bacillus subtilis ( Bacillus subtilis ) The preparation method of the bacteriocin crude product that can inhibit Gram-positive and Gram-negative bacteria produced by HJD A32 first uses 1 part of glucose, 1 part of yeast extract, and 100 parts of distilled water for fermentation. The value is 7.2-7.4, the Bacillus subtilis ( Bacillus subtilis ) The HJD A32 strain was inoculated into the culture medium, and the culture condition was 36°C for 48 hours to obtain a fermentation broth; then the fermentation broth was centrifuged at 10,000 r / m for 15 minutes and the supernatant was taken, and ammonium sulfate solid powder was slowly added to the supernatant for Salt out, stir while adding until the saturation reaches 80%, then stand at 4°C for 12h, then centrifuge at 12000 r / m, 5°C for 30 min, discard the supernatant to obtain the crude product of bacteriocin precipitation.

[0037] The obtained crude bacteriocin precipitate was dissolved in 1 / 40 volume of 0.02mol / L sodium ac...

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Abstract

A bacillus subtilis HJDA32 strain is preserved on September 25, 2012 with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.6624, which is separated from solid fermentative substrate of vinegar in the Shanxi province Qingxu mature vinegar plant. The strains are cultured in a beef extract peptone basic medium and arranged in pole, single, paired or chained shape with the size between 0.6-0.85mu m*1.5-3.5mu m, and have capsules; elliptical spores are cultured on a raw spore culture medium; the strain is identified to be bacillus subtilis through catalase and carbon source utilization test, gelatin hydrolysis, starch hydrolysis, ethanol oxidation, acetic acid oxidation and other physiological and biochemical identifications and 16SrDNA (deoxyribonucleic acid) phylogenetic analysis; and the bacillus subtilis HJDA32 strain is used as a production strain, cultured with fermenting culture fluid, and subjected to centrifuge, ammonium sulfate powder salting out and centrifuge to obtain the bacteriocin. The bacteriocin is wide in antimicrobial spectrum and high in bacteriostatic activity, can inhibit various food-borne pathogenic bacteria, as well as gram positive and gram negative bacteria causing food spoilage, and can be used in food preservatives.

Description

technical field [0001] The invention relates to the field of microbes, in particular to a new strain of Bacillus subtilis and a preparation method of the bacteriocin produced by it, which can inhibit Gram-positive and Gram-negative bacteria. Background technique [0002] Food safety is an important topic worldwide. At present, one of the main methods to solve food safety is to add chemical preservatives, but long-term studies have found that excessive consumption of preservatives has adverse effects on blood pressure, heart, kidneys, etc., and even some chemical preservatives are carcinogenic, teratogenic and easily cause problems such as food poisoning. Thermal sterilization is also one of the important methods to kill microorganisms in food processing, but for some low-temperature products, such as low-temperature meat products, if the heating temperature is too high, the color and flavor will change significantly, and the nutritional value will also be greatly reduced. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P21/00C12R1/125
Inventor 郝林贾丽艳段先瑶
Owner SHANXI AGRI UNIV
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