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Baltic shewanella oneidensis bacteriophage SppYZU01 and application thereof

A technology of Shewanella and bacteriophage, applied in the field of bioengineering, can solve the problems of high preparation cost, continuous pollution, changing the color and flavor of food, etc., and achieve the effects of no toxic and side effects, reducing cross-contamination, and prolonging the storage period.

Active Publication Date: 2018-02-23
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these bacteriostatic agents can meet the needs of customers for food safety, they have defects such as poor fresh-keeping effect in low-temperature environments, high preparation costs, and changes in the color and flavor of food.
In addition, Shewanella forms a biofilm on the surface of food and processing utensils during the process of adapting to the natural environment. The bacteria in the film are hundreds of times more resistant to conventional preservatives and antibacterial agents, making it difficult to remove. This will lead to continuous pollution and bring serious harm to the processing and preservation of aquatic products and meat products

Method used

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  • Baltic shewanella oneidensis bacteriophage SppYZU01 and application thereof
  • Baltic shewanella oneidensis bacteriophage SppYZU01 and application thereof
  • Baltic shewanella oneidensis bacteriophage SppYZU01 and application thereof

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Experimental program
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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. 30 mL of sewage sample is put into a 50 mL centrifuge tube, centrifuged at 5000 × g for 10 min, 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. For Shewanella Baltica SYZU01, culture overnight on a shaker at 25°C. On the next day, transfer the culture in the test tube to a sterile centrifuge tube, centrifuge at 5000×g for 10 min at 4°C, and pass the supernatant through a 0.22 μm filter membrane. The phage stock solution was obtained and stored in a refrigerator at 4°C.

[0037] Dilute the phage stock solution with sterile SM buffer, take 100 μL of appropriate dilution of phage solution and 100 μL of Shewanella balticii SYZU01 in logarithmic growth phase, mix at room temperature for 10 minutes, then ...

Embodiment 2

[0049] Embodiment 2, the inhibitory effect of phage SppYZU01 to the biofilm of Shewanella Baltica

[0050] Take Shewanella Balticae SYZU01 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 SppYZU01 suspension (10 4 PFU / mL, 10 5 PFU / mL, 10 6 PFU / mL, 10 7 PFU / mL, 10 8 PFU / mL) each 150 μL; 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-well plate with sterile PBS until...

Embodiment 3

[0052] Embodiment 3, the bacteriostasis of bacteriophage SppYZU01 in fish juice storage process

[0053] 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. Press Shewanella Baltica SYZU01 by 10 6 CFU / mL concentration was inoculated into 30mL fish juice, and 30 μL phage SppYZU01 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 and a 4 °C refrigerator for cons...

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Abstract

The invention relates to Baltic shewanella oneidensis bacteriophage SppYZU01 and application thereof. The Baltic shewanella oneidensis bacteriophage SppYZU01 has the preservation number CCTCC NO:M 2016715. The invention also discloses application of the bacteriophage in food preservation and food processing as a biological bacteriostatic agent. The Baltic shewanella oneidensis bacteriophage SppYZU01 provided by the invention has broad-spectrum and efficient cracking activity, culture and suspension thereof can be taken as a preservative and used for controlling growth of shewanella oneidensisand formation of a biofilm during food cold storage, and shelf life of a product is effectively prolonged.

Description

technical field [0001] The invention relates to a Baltic Shewanella phage isolate SppYZU01, and its antibacterial application in food production and preservation as a food additive or biological antibacterial agent, especially capable of inhibiting refrigerated freshwater or seawater products from the Baltic Sea The spoilage caused by the proliferation of Shewanella prolongs the shelf life of products. Belongs to the field of bioengineering. Background technique [0002] Shewanella baltica is the main psychrophilic spoilage bacterium in aquatic products and meat products. It grows fast and has strong spoilage ability under low temperature storage (0-5°C), and finally forms the dominant bacterial group in the spoilage process. , and produce histamine, sulfur, indole, aldehyde, trimethylamine and other metabolites, resulting in deterioration of sensory quality, causing serious economic losses to aquatic products and meat processing industries. Effectively inhibiting the repr...

Claims

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Application Information

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IPC IPC(8): C12N7/00A23B4/22C12R1/92
Inventor 杨振泉张辉高璐饶胜其陶晓雅
Owner YANGZHOU UNIV
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