Paenibacillus sp. bacteriocin crude extract as well as preparation method and application thereof

A technology of Paenibacillus and Bacillus, which is applied in the field of Paenibacillus bacteriocin crude extract and its preparation, can solve the problems of poor bacteriocin stability, food safety risks, complicated preparation process, etc., and achieves excellent application value and high food safety. performance, cost reduction

Active Publication Date: 2015-07-15
BRIGHT DAIRY & FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Therefore, the technical problem to be solved by the present invention is to aim at the current situation of poor stability of existing bacteriocins, high preparation cost, insufficient sources, diverse fermentation medium components, complex formula, cumbersome preparation process and potential food safety risks, and provides A kind of crude extract of Paenibacillus bacteriocin and its preparation method and application

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  • Paenibacillus sp. bacteriocin crude extract as well as preparation method and application thereof
  • Paenibacillus sp. bacteriocin crude extract as well as preparation method and application thereof
  • Paenibacillus sp. bacteriocin crude extract as well as preparation method and application thereof

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Embodiment 1

[0029] The preparation of embodiment 1 Paenibacillus bacteriocin crude extract and the detection of antibacterial activity

[0030] 1. Materials and methods

[0031] Preparation of seeds (fermentation strains): freeze-drying of Paenibacillus sp. (the preservation number of Paenibacillus sp. is CGMCC No.8333, and the source of the strain is referred to the Chinese patent publication number CN103740618A). Dissolve the powder in a small amount of sterile distilled water, use an inoculation loop to draw a line 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 with a 2% (volume percentage) inoculum Culture medium (purchased from...

Embodiment 2

[0044] The preparation of embodiment 2 Paenibacillus bacteriocin crude extract and the detection of antibacterial activity

[0045] 1, material and method: with embodiment 1.

[0046] 2. Preparation of Paenibacillus bacteriocin crude extract

[0047] 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 96 hours to obtain a fermentation liquid, and the above fermentation liquid 8 Centrifuge at 10,000g for 10min to take the supernatant, slowly add ammonium sulfate to the supernatant, and centrifuge again at 10,000g for 10min to collect the precipitate precipitated when the saturation of ammonium sulfate is 50-70%. The 1000Da dialysis bag was dialyzed for 1 day, and the water was changed 4 times a day. After the dialysis was completed, the solution in the bag was freeze-dried to obtain the crude extract B of bacteriocin. Wherein, the formula of th...

Embodiment 3

[0053] The preparation of embodiment 3 Paenibacillus bacteriocin crude extract and the detection of antibacterial activity

[0054] 1, material and method: with embodiment 1.

[0055] 2. Preparation of Paenibacillus bacteriocin crude extract

[0056] Inoculate the Paenibacillus sp. CGMCC No.8333 strain into a sterile medium with a 2% (volume percentage) inoculum, and ferment at 30° C. and 180 rpm for 72 hours to obtain a fermented liquid. The above-mentioned fermented liquid, 4 Centrifuge at 15,000g for 5 minutes to take the supernatant, slowly add ammonium sulfate to the supernatant, and centrifuge again at 15,000g for 5 minutes to collect the precipitate precipitated when the saturation of ammonium sulfate is 40-60%. The 1000Da dialysis bag was dialyzed for 2 days, and the water was changed 3 times a day. After the dialysis was completed, the solution in the bag was freeze-dried to obtain the crude extract C of bacteriocin. Wherein, the formula of the fermentation medium i...

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Abstract

The invention discloses a paenibacillus sp. bacteriocin crude extract as well as a preparation method and application thereof. The preparation method comprises the following steps: (1) inoculating paenibacillus sp. into a culture medium for culturing the paenibacillus sp., shocking and carrying out fermentation cultivation to obtain fermentation liquor, and centrifuging the obtained fermentation liquor so as to take liquid supernatant; and (2) adding ammonium sulfate to the obtained liquid supernatant, centrifuging and collecting precipitates which are separated out when the saturation of the ammonium sulfate is 30-70%, dissolving the obtained precipitates and then dialyzing, and carrying out freeze drying to obtain the paenibacillus sp. bacteriocin crude extract. Raw materials for preparing the paenibacillus sp. bacteriocin crude extract are bran, wide in source and low in cost, and a fermentative strain is a paenibacillus sp. single strain; and the technology facilitates the standardization of product quality and the cost control of industrialized mass production. The paenibacillus sp. bacteriocin crude extract can be directly applied to food production, thus achieving the purposes of inhibiting the proliferation of putrefying bacteria and prolonging the shelf period of food and reducing the application cost.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a crude extract of Paenibacillus bacteriocin and its preparation method and application. 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 condition...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00A23L1/305C12R1/01
Inventor 韩瑨吴正均刘振民郭本恒吴江
Owner BRIGHT DAIRY & FOOD
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