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Preparation method and application of paenibacillus bacteriocin

A technology of Paenibacillus and Bacillus, applied in the field of preparation of Paenibacillus bacteriocin preparations, can solve the problems of high preparation cost, poor stability of bacteriocin preparations, insufficient sources, etc., achieve stable properties, prolong food shelf life, and inhibit proliferation Effect

Active Publication Date: 2015-07-08
BRIGHT DAIRY & FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Therefore, the technical problem to be solved by the present invention is to provide a preparation method and application thereof for the existing bacteriocin preparations with poor stability, high preparation costs and insufficient sources.

Method used

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  • Preparation method and application of paenibacillus bacteriocin
  • Preparation method and application of paenibacillus bacteriocin
  • Preparation method and application of paenibacillus bacteriocin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The preparation of embodiment 1 Paenibacillus bacteriocin preparation and the detection of antibacterial activity

[0025] 1. Materials and methods

[0026] Preparation of seeds (fermentation strains): freeze-dried Paenibacillus sp. Dissolve the powder with a small amount of sterile distilled water, use an inoculation loop to take a ring and streak it on TYC solid medium (purchased from OXOID Co, UK), take it out after 48 hours of aerobic culture at 30°C, and use an inoculation loop to pick a single colony and put it into 10mL TYC liquid Culture medium (purchased from OXOID Co, UK), use a vortex shaker to evenly disperse the colonies in the liquid medium, take it out after 24 hours of shaking culture at 30°C and 180rpm, and inoculate it in TYC liquid medium with an inoculum of 2% (volume percentage) (purchased from OXOID Co, UK), after shaking culture at 30°C and 180rpm for 24 hours, the culture was centrifuged at 15,000rpm for 10 minutes, the supernatant was discarded...

Embodiment 2

[0039] The preparation of embodiment 2 Paenibacillus bacteriocin preparation and the detection of antibacterial activity

[0040] 1, material and method: with embodiment 1.

[0041] 2. Preparation of Paenibacillus bacteriocin preparation

[0042] Inoculate the Paenibacillus sp. CGMCC No.8333 strain into a sterile fermentation medium with an inoculum size of 1% (volume percentage), 37 ° C, 300 rpm shaking fermentation culture for 48 hours to obtain a fermentation liquid, and the above fermentation liquid 10,000 Centrifuge at rpm for 10 min to take the supernatant, add absolute ethanol with 4 times the volume of the supernatant, mix well and let stand at 2°C for 12 hours for alcohol precipitation, then centrifuge at 8°C and 10,000 rpm for 10 min to take the supernatant, Paenibacillus bacteriocin preparation B was obtained after concentrated under pressure and freeze-dried. Among them, the formula of the fermentation medium is: yeast extract 1%, casein hydrolyzate 2%, sucrose 1...

Embodiment 3

[0048] The preparation of embodiment 3 class bacillus bacteriocin preparation and the detection of antibacterial activity

[0049] 1, material and method: with embodiment 1.

[0050] 2. Preparation of Paenibacillus bacteriocin preparation

[0051] Inoculate the Paenibacillus sp. CGMCC No.8333 strain into a sterile fermentation medium with an inoculum size of 2% (volume percentage), 30°C, 180rpm shaking fermentation culture for 30h to obtain a fermentation liquid, and the above fermentation liquid 15,000 Centrifuge at rpm for 5 minutes to take the supernatant, add absolute ethanol that is 3 times the volume of the supernatant, mix well and let stand at 8°C for 24 hours for alcohol precipitation, centrifuge again at 15,000rpm for 5 minutes to take the supernatant, concentrate under reduced pressure and freeze Paenibacillus bacteriocin preparation C was obtained after drying. Among them, the formula of the fermentation medium is: yeast extract 0.5%, peptone 1.5%, sucrose 5%, L-...

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Abstract

The invention discloses a preparation method of paenibacillus bacteriocin. The preparation method comprises the following steps: (1) inoculating the paenibacillus into a culture medium for culturing the paenibacillus, culturing in oscillating and fermenting to obtain fermented liquor, centrifuging the obtained fermented liquor, and taking the supernatant; (2) mixing the obtained supernatant with ethanol in volume ratio of (2:1)-(4:1), standing obtained mixed liquor for 12-36 hours at temperature of 2-10 DEG C, centrifuging the mixed liquor to collect precipitate, removing a solvent to obtain the paenibacillus bacteriocin. Compared with the traditional bacteriocin, the bacteriocin prepared through the method disclosed by the invention is more stable, the bacteriocin preparation can be directly applied to the production of foods, thereby achieving the aim of inhibiting the proliferation of putrefying bacteria and prolonging shelf life of the food, and then the application cost of the food preservative is reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method and application of a Paenibacillus bacteriocin preparation. Background technique [0002] Bacteriocins are substances with antibacterial activity produced by certain bacteria, and their main components are polypeptides, proteins or protein complexes, which were first proposed by Jacob in 1953. In recent years, the negative effects brought about by the popularization and even abuse of broad-spectrum antibiotics have become increasingly prominent, and the research on new antibiotic substitutes is imminent. Bacteriocins, as protein substances, are highly safe because they can be degraded by proteases, so they have received more and more attention. Although the currently developed bacteriocin (Nisin) has a very high antiseptic and antibacterial application value, due to the characteristics of Nisin itself (stable properties under low pH conditions, and g...

Claims

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Application Information

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IPC IPC(8): C12P21/00A23L3/3463C12R1/01
Inventor 吴正均郭本恒刘振民韩瑨徐晓芬
Owner BRIGHT DAIRY & FOOD
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