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Shewanella putrefaciens phage and application thereof

A technology of Shewanella putrefaciens and phages, applied in the direction of phages, viruses/phages, meat/fish preservation with chemicals, etc., can solve the hazards of aquatic products and meat products processing and preservation, poor low-temperature preservation effect, and sustainable Pollution and other issues to achieve the effect of maintaining freshness, reducing corruption and reducing pollution

Active Publication Date: 2018-03-23
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these antibacterial agents can inhibit the growth of Shewanella putrefaction, they often have poor low-temperature preservation effects and high preparation costs.
In addition, during the growth process of Shewanella putrefaciens, it is very easy to form a biofilm on the surface of food and processing utensils. The resistance of the bacteria in the film to conventional preservatives and antibacterial agents can be increased hundreds of times, and it is difficult to be removed. , resulting in continuous pollution and serious harm to the processing and preservation of aquatic products and meat products

Method used

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  • Shewanella putrefaciens phage and application thereof
  • Shewanella putrefaciens phage and application thereof
  • Shewanella putrefaciens phage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1, phage isolation and purification preparation

[0035] Phage isolation

[0036] The sample of the present invention is collected from the sewage of farmers' market in Yangzhou, Jiangsu, and 30 mL of sewage sample is put into a 50 mL centrifuge tube, centrifuged at 5000 × g for 10 min, and 5 mL of supernatant is added to 5 mL of LB liquid medium, and 100 μL of logarithmic growth phase is added at the same time. Shewanella putrefaciens CICC 22940, cultured overnight on a shaker at 25°C, the next day, transfer the culture in the test tube to a sterile centrifuge tube, centrifuge at 5000×g for 10min at 4°C, take the supernatant and pass through a 0.22μm filter membrane , to obtain the phage stock solution, and store it in a refrigerator at 4°C.

[0037] Doubling dilution of the phage stock solution with sterile SM buffer, take 100 μL of the appropriate dilution of the phage solution and 100 μL of Shewanella putrefaciens CICC 22940 in logarithmic growth phase a...

Embodiment 2

[0048] Embodiment 2, the inhibitory effect of phage SppYZU05 to Shewanella putrefaciens biofilm

[0049] Take Shewanella putrefaciens (CICC 22940+ATCC BAA-1097, 1:1 mixed) in the logarithmic growth phase and dilute it with nutrient broth medium, and the final concentration is 1×10 7 CFU / mL, experimental group: add diluted bacterial solution (10 7 CFU / mL) and phage SppYZU05 suspension (10 4 PFU / mL, 10 5 PFU / mL, 10 6 PFU / mL, 10 7 PFU / mL, 10 8 PFU / mL) each 150μL; control group: add diluted bacterial solution (10 7 CFU / mL) and 150 μL of nutrient broth medium; blank group: add 300 μL nutrient broth medium to each well; incubate at 37°C for 24 hours; take out the 96-well plate, suck out the suspended bacteria, and wash the 96-well plate with sterile PBS 3 Once, blow dry to fully remove planktonic cells; add 200 μL of crystal violet staining solution with a concentration of 0.2% to each well, and stain for 30 minutes; suck out the staining solution, and thoroughly wash the 96-w...

Embodiment 3

[0051] Embodiment 3, the bacteriostasis of bacteriophage SppYZU05 in fish juice storage process

[0052] After the fresh grass carp is slaughtered, take the meat on both sides of the back of the fish, peel it, cut it into thin strips, and weigh it. The ratio of the volume of water to the mass of fish meat is 2:1. After adding water and boiling for 5 minutes, filter it with gauze, add water again, and recover To the amount added for the first time, continue to boil for 5 minutes, filter with filter paper, adjust the pH of the filtrate to 7 with NaOH, add 40mg L-cysteine ​​and 40mg L-methionine to each liter of fish juice, and divide into cones In a shaped bottle, sterilize at 121°C for 15 minutes. Put Shewanella putrefaciens (CICC 22940+ATCC BAA-1097, 1:1 mix) in 10 6 CFU / mL concentration was inoculated into 30mL fish juice, and 30 μL phage SppYZU05 suspension (10 8 PFU / mL), 30 μL of SM buffer was added to the control group, and the inoculum was placed in a 25 °C incubator an...

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Abstract

The invention belongs to the field of bioengineering, and relates to a shewanella putrefaciens phage isolated strain and application thereof. The shewanella putrefaciens phage isolated strain is shewanella putrefaciens phage SppYZU05 with the preservation number being CCTCC NO:M2016717. The invention also discloses bacteria inhibition application of the shewanella putrefaciens phage SppYZU05 as afood additive composition or a biological fresh-keeping agent in food production and storage, particularly, spoilage of fresh water or sea water products in cold chain circulation caused by shewanellaputrefaciens phage multiplication can be inhibited, and the product shelf life is prolonged.

Description

technical field [0001] The present invention relates to a bacteriophage isolate of Shewanella putrefaciens and its antibacterial application in food production and storage as a food additive or biological preservative, especially capable of inhibiting spoilage in fresh water or seawater products in cold chain circulation The spoilage caused by the proliferation of Shewanella prolongs the shelf life of products. Belongs to the field of bioengineering. Background technique [0002] Shewanella putrefaciens is a specific spoilage bacterium in the cold chain circulation of high-moisture protein foods such as aquatic products and meat products. It grows fast under low-temperature storage (0-8°C) conditions, has strong spoilage ability, and eventually forms spoilage. The dominant flora in the process, and produce H 2 S. Reduction of TMAO will lead to deterioration of sensory quality such as sticky meat quality and color change, which will cause serious economic losses to aquatic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23B4/22
CPCA23B4/22C12N7/00C12N2795/10321
Inventor 杨振泉陶晓雅张辉高璐饶胜其
Owner YANGZHOU UNIV
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