Active strain for inhibiting drug-resistant food-borne pathogenic bacteria and application thereof

A technology of food-borne pathogenic bacteria and pathogenic bacteria, applied in the field of microbiology, can solve problems such as damage to human health, impact on food quality, food preservative effect and safety doubts, and achieve broad-spectrum antibacterial activity and strong antibacterial activity effect of effect

Active Publication Date: 2022-03-25
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Due to the many disadvantages brought about by the long-term use of chemical preservatives, such as affecting the original quality of food and damaging human health, the effect and safety of food preservatives have been questioned
At the same time, the massive abuse of antibiotics has made drug-resistant strains in food a major hidden danger to human health.

Method used

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  • Active strain for inhibiting drug-resistant food-borne pathogenic bacteria and application thereof
  • Active strain for inhibiting drug-resistant food-borne pathogenic bacteria and application thereof
  • Active strain for inhibiting drug-resistant food-borne pathogenic bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1, 16S rDNA sequence analysis and phylogenetic tree construction of Pseudomonas fluorescens S-541

[0037]In the present invention, an active bacterial strain is isolated from vegetable garden soil in Baiyun District, Guangzhou City, Guangdong Province as a strain to be tested, and DNA is extracted from the active bacterial strain.

[0038] Extraction of the total DNA of the strain: Scrape an appropriate amount of the strain to be tested from the NA slant medium preserved at low temperature (4°C) in the laboratory, inoculate it in 100ml liquid medium, cultivate it at 28°C for 72 hours, and then use bacterial genome DNA extraction reagent Kit (BIOMIGA Company, USA) was used to extract genomic DNA, and the operation steps were carried out according to the instructions of the kit. The obtained genome was electrophoresed with 0.8% agarose, and the extraction effect was detected on a gel imager and used for genome sequencing.

[0039] The cloning primers of the gene ...

Embodiment 2

[0042] Embodiment 2, the antibacterial activity assay of Pseudomonas fluorescens (Pseudomonas fluorescens) S-541 fermentation supernatant

[0043] 1) Obtaining the fermentation supernatant of Pseudomonas fluorescens S-541:

[0044] Pseudomonas fluorescens S-541 preserved on NA agar slant was inoculated into NB liquid medium for activation, and cultured at 37° C. on a shaker at 160 r / min for 24 hours. An appropriate amount of activated Pseudomonas fluorescens (Pseudomonas fluorescens) S-541 was inoculated in 100 mL of MH liquid medium according to the inoculum size of 10%, cultured at 37° C. on a shaking table at 160 r / min for 72 h. The cultured strain fermentation liquid was centrifuged at 8000 r / min at 4° C. for 10 min to obtain the fermentation supernatant.

[0045] 2) Antibacterial activity test of Pseudomonas fluorescens (Pseudomonas fluorescens) S-541 fermentation supernatant:

[0046] Take food-borne pathogenic bacteria and corresponding drug-resistant strains as indic...

Embodiment 3

[0052] Example 3, Pseudomonas fluorescens (Pseudomonas fluorescens) active product extraction, isolation and identification of S-541

[0053] Cultivate Pseudomonas fluorescens S-541 with MH solid medium for 2-4 days, extract with methanol for 24 hours, and then use a rotary evaporator to concentrate under reduced pressure at 40°C to obtain a crude extract with antibacterial active substances . The crude extract was dissolved in 5ml of methanol, filtered through a 0.22 μm membrane, separated and purified by semi-preparative liquid chromatography, the chromatographic conditions were as follows, instrument: Shimadzu LC-20A, chromatographic column: Waters C18 column (20mm×250mm, 5 μm), with volume fraction 15% methanol and ultrapure water as the mobile phase, the flow rate is 3mL / min, to obtain two parts of components A and B, such as image 3 .

[0054] Adopt antibacterial activity tracking, show good antibacterial activity in component A, therefore, adopt the same method to fu...

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Abstract

The invention relates to the technical field of microorganisms, and particularly discloses an active strain for inhibiting drug-resistant food-borne pathogenic bacteria and application. The 16S rDNA sequence of the active strain for inhibiting the drug-resistant food-borne pathogenic bacteria is as shown in SEQ ID NO: 1. The active strain is Pseudomonas fluorescens and is preserved in the Guangdong Microbial Culture Collection Center, and the preservation number of the active strain is GDMCC No: 61981. The antibacterial active substance metabolized by the pseudomonas fluorescens disclosed by the invention has broad-spectrum antibacterial activity, particularly has very strong antibacterial activity on drug-resistant staphylococcus aureus, and can be used as a potential food preservative to be applied to food preservation.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to an active bacterial strain for inhibiting drug-resistant food-borne pathogenic bacteria and its application. Background technique [0002] Food safety is the basis for the harmony, stability, development and progress of human society. Our country clearly proposes that building a healthy China in an all-round way will become the future development direction, and attaching importance to food safety has become the focus of attention of the whole society. At this stage, food safety problems are still breaking out frequently around the world. Foodborne diseases caused by microorganisms are recognized as the number one food safety problem. 70% of children who die of food poisoning are caused by pathogenic microorganisms. Food spoilage accounts for more than 10% of food output value, and the prevention and control of pathogenic microorganisms is a major demand for food safety. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/6409A23L3/3508A23L3/3571C12R1/39
CPCC12P7/6409A23L3/3508A23L3/3571Y02A50/30
Inventor 柏建玲陈玲何莹龙王惠惠赵辉吴清平张菊梅陈谋通谢新强蔡淑珍丁郁莫树平
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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