Bacteriocin, and preparation method and application thereof

A technology of bacteriocin and Lactobacillus sake, which is applied in the field of bacteriocin, can solve the problems of unsatisfactory purification effect and yield, and achieve the effects of avoiding denaturation and inactivation, high yield, and good separation and purification

Inactive Publication Date: 2015-10-21
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the protein separation and purification method is also applicable to the separation and purification of lactic acid bacteria bacteriocins, but according to the characteristics of different bacteriocins, the purification methods of bacteriocins are also different, and the purification effect and yield disclosed so far are not very ideal

Method used

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  • Bacteriocin, and preparation method and application thereof
  • Bacteriocin, and preparation method and application thereof
  • Bacteriocin, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Isolation of Lactobacillus sake LZ217

[0045] Take fresh milk and dilute it with 0.9% physiological saline in 10-fold gradients, absorb 0.3ml of each dilution gradient and spread it on a plate containing Lactobacillus sake LZ217 screening medium, and incubate in an anaerobic manner at 37°C for 36-48 hours. Select colonies with good growth and different shapes and large calcium-dissolving circles for numbering, and repeatedly isolate and purify by plate streaking separation method, pick a single colony of Lactobacillus sake LZ217 on the plate, inoculate it into MRS liquid medium, and culture at a constant temperature of 37°C After 24 hours, the strains were numbered and preserved, and the strains were preserved at -80°C.

[0046] Screening of Lactobacillus sake LZ217 producing antibacterial active metabolites

[0047] Three rings of the isolated acid-producing strains were picked and inoculated into MRS liquid medium, and cultured at 37°C for 24 hours. Centrifuge at 4...

Embodiment 2

[0070] Embodiment 2: the preparation of bacteriocin

[0071] Lactobacillus sake LZ217 was inoculated in MRS liquid medium, and cultured at 37°C for 12 hours to obtain a seed liquid; inoculated at 5% inoculum size in MRS liquid medium, cultured at 37°C for 24 hours, and centrifuged to remove bacteria. Store at 4°C for later use. The centrifugation conditions were 12000 rpm, 20 min, and 4°C.

[0072] The crude extract of bacteriocin was obtained by centrifugation, and the bacteriocin was named as sake lactobacillus LZ217.

Embodiment 3

[0074] Separation and Purification of Bacteriocins (1)

[0075] The fermentation supernatant was roughly extracted by macroporous resin, and the specific experimental steps were as follows

[0076] (1) Weigh 30g of macroporous resin XAD-1180, soak it in absolute ethanol for 24 hours to fully swell and activate it, then fully rinse it with deionized water, and finally filter it with a suction filter bottle for later use.

[0077] (2) Pack the treated macroporous resin into a 2.5×30 cm glass chromatography column by a wet method, avoiding the entry of air bubbles as much as possible.

[0078] (3) 1L of fermentation supernatant was passed through the column, and the flow rate was controlled at 10mL / min. The duration is about 100min.

[0079] (4) Wash the chromatography column with 500 mL of ultrapure water to remove the residual culture solution attached to the surface of the resin.

[0080] (5) Use 200mL20% methanol solution to elute the chromatography column, the flow rate i...

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Abstract

The invention provides a bacteriocin, and a preparation method and an application thereof. The bacteriocin belongs to broad spectrum bacteriocin, and an antibacterial spectrum comprises Gram-negative bacteria and Gram-positive bacteria, such as Pseudomonas putida, Bacillus subtilis, Listeria monocytogenes, Shigella flexneri, Staphylococcus citreus, Escherichia coli and salmonella.

Description

technical field [0001] The invention relates to a bacteriocin produced by a strain of Lactobacillus sakei LZ217 (Lactobacillus sakei LZ217) and the application of the microorganism to produce the bacteriocin. Background technique [0002] Bacteriocin is a short antibacterial peptide produced by Lactobacillus, which is high temperature resistant, acid resistant, degradable in the human body, safe and non-toxic, and has strong antibacterial activity. Bacteriocin is a kind of protein or low molecular weight polypeptide substance with antibacterial activity produced by microorganisms through the ribosome synthesis mechanism. The antibacterial range is not limited to homologous bacteria, and the producing bacteria are autoimmune to their bacteriocins. [0003] As a special antibacterial and antibacterial protein, bacteriocin has the characteristics of strong antibacterial activity, good biocompatibility, stable performance, biodegradability, digestibility and high biological safe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/335C12P21/00C07K1/22C07K1/20C07K1/18A23L3/3526A23K3/00A61K38/02A61P31/04
CPCA23L3/3526A61K38/02C07K14/335Y02A50/30
Inventor 顾青顾容铖宋达峰沈雷
Owner ZHEJIANG GONGSHANG UNIVERSITY
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