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Bacteriocin PE-ZYB1 with preservation and fresh-keeping effects for food and application thereof

A PE-ZYB1, food preservation technology, applied in the biological field

Inactive Publication Date: 2020-02-04
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the commonly used commercial lactobacillus nisin has antibacterial effect only on Gram-positive bacteria, it is urgent to find more lactobacillus with broad antibacterial spectrum and strong antibacterial activity.

Method used

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  • Bacteriocin PE-ZYB1 with preservation and fresh-keeping effects for food and application thereof
  • Bacteriocin PE-ZYB1 with preservation and fresh-keeping effects for food and application thereof
  • Bacteriocin PE-ZYB1 with preservation and fresh-keeping effects for food and application thereof

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Experimental program
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Effect test

Embodiment 1

[0045] The separation and purification of embodiment 1 bacteriocin PE-ZYB1

[0046] (1) Preparation of lactic acid bacteria zy-B fermentation supernatant and assay method of bacteriocin titer

[0047] Transfer the lactic acid bacteria zy-B preserved on the slant to the MRS liquid medium, place it on a constant temperature shaker and culture it at 28°C and 150r / min, and inoculate it into 1L MRS liquid medium according to the inoculation amount of 3% after activation for 2 generations , cultured at 28°C for 48h, collected the fermentation broth and centrifuged at 12,000rpm and 4°C for 10min, removed the bacteria and retained the fermentation supernatant, filtered the fermentation supernatant with a 0.22μm filter membrane, and filtered the fermentation supernatant with Listeria Bacteria were used as indicator bacteria for the determination of antibacterial activity and stored at 4°C for later use. Among them, the determination of antibacterial activity adopts the double-layer ag...

Embodiment 2

[0064] Molecular mass determination and amino acid analysis of embodiment 2 bacteriocin PE-ZYB1

[0065] Send the purified bacteriocin PE-ZYB1 to Beijing Baitai Paike Biotechnology Co., Ltd., and use LC-MS / MS system (Shimadzu LC-20AD nano flow-LC and Thermo Fisher Scientific Q Exactive massspectrometer) to analyze the polypeptide fragments. analyze. The scanning range of the primary mass spectrometer is 350-1800 (m / z), the resolution is 70000, and the resolution of the secondary MS / MS is 17500. The obtained amino acid sequence was compared with the sequences preserved in the Universal Protein Database (UniProt), BLAST (http: / www.uniprot.org / BLAST / ) and Anti-Peptide Database (APD) by MaxQuant analysis software (http: / aps.unmc .edu / ap / main.html).

[0066] LC-MS / MS analysis found that the bacteriocin PE-ZYB1 has the amino acid sequence shown in SEQ ID NO: 1 ( Image 6 ), the molecular weight of bacteriocin PE-ZYB1 is 2019.22Da. Compared with the universal protein database (Un...

Embodiment 3

[0067] The bacteriostatic spectrum determination of embodiment 3 bacteriocin PE-ZYB1

[0068] Bacteriocin PE-ZYB1 solution (diluted to 50 μg / mL with PBS) was carried out using the double-layer agar Oxford cup method, with the indicator bacteria in Table 1, to carry out the indicator bacteria inhibition spectrum experiment, and measure the diameter of the inhibition zone. For the specific operation method of the Oxford cup method, refer to "Jiao Dongdong, Ren Wenbin, Zhao Yamei, et al. Isolation and diversity analysis of antibacterial active lactic acid bacteria in the intestines of animals in the South China Sea [J]. Food Science, 2017(08):82-87 "." Among them, Listeria monocytogenes was inoculated on TSA-YE medium, Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Bacillus cereus, Enterobacter aerogenes, Pseudomonas aeruginosa, Streptococcus hemolytics, Proteus bacillus were inoculated on nutrient agar medium, Vibrio parahemolyticus was inoculated on nutrient agar m...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to bacteriocin PE-ZYB1 with preservation and fresh-keeping effects for food and application thereof. The bacteriocin PE-ZYB1 is obtained by separation and purification from lactic acid bacteria zy-B by a four-step method using ethyl acetate extraction, Sephadex LH-20 column chromatography system purification, ion exchange chromatography purification and semi-preparative reversed-phase liquid chromatography purification. The bacteriocin PE-ZYB1 has an amino acid sequence as shown in SEQ ID NO: 1, and is similar toan antibacterial peptide Andersonin-X1 with similarity of only 38.09%. The bacteriocin PE-ZYB1 has broad-spectrum antibacterial activity against both gram-positive and gram-negative food-borne pathogens and spoilage bacteria, has high heat resistance and stability at high temperature, can still maintain good bacteriostatic activity at a pH value of 2-7, especially has a killing effect on Listeriamonocytogenes, and can be used in the field of food preservation and fresh-keeping.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a bacteriocin PE-ZYB1 with food antiseptic and fresh-keeping functions and an application thereof. Background technique [0002] During the process of food processing and preservation, it is very easy to be spoiled by microbial contamination. Microbial contamination has become a worldwide food safety problem. It not only causes food spoilage, but also causes significant health and economic losses. For example, Listeria monocytogenes (Listeria monocytogenes), Salmonella typhimurium, Escherichia coli, Campylobacter, etc. often lead to severe listeriosis, diarrhea syndrome, hemolytic uremia and salmonellosis. of foodborne diseases. At present, the use of preservatives to inhibit microorganisms and delay spoilage is one of the important technologies for food preservation. However, chemical preservatives are the most used in food, which affect human health, and their applica...

Claims

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Application Information

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IPC IPC(8): C07K7/08A23L3/3526
CPCA23L3/3526C07K7/08
Inventor 刘颖徐春厚张瀛孙力军王雅玲房志家邓旗
Owner GUANGDONG OCEAN UNIVERSITY
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