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CN-121970804-A - Salmonella phage nanoemulsion preparation and preparation method and application thereof

CN121970804ACN 121970804 ACN121970804 ACN 121970804ACN-121970804-A

Abstract

The invention discloses a salmonella phage nanoemulsion preparation, a preparation method and application thereof, wherein the salmonella phage nanoemulsion preparation mainly comprises, by mass, 5% -15% of salmonella phage suspension, 84% -94% of edible oil, 0.5% -3.0% of Pickering stabilizer dispersion liquid and 0% -2.0% of protective agent. The invention also discloses a preparation method of the salmonella phage nanoemulsion preparation and application of the salmonella phage nanoemulsion preparation in bacteriostasis and fresh-keeping of meat products. The salmonella phage nanoemulsion preparation has a more essential stabilization mechanism, a more stable system, better phage activity maintenance, better food applicability, safety and friendliness, longer bacteriostasis effect and application of being more suitable for meat product preservation.

Inventors

  • ZHONG LE
  • YUAN DI
  • SUN YUNQIAN
  • ZHOU JINXIAN
  • LIANG WEIQUAN
  • CHEN YAXIN
  • CHENG ZHENG
  • XIAO NAIYU
  • ZHONG YUNYUN

Assignees

  • 仲恺农业工程学院

Dates

Publication Date
20260505
Application Date
20260206

Claims (10)

  1. 1. A salmonella phage nanoemulsion preparation is characterized by mainly comprising, by mass, 5% -15% of salmonella phage suspension, 84% -94% of edible oil, 0.5% -3.0% of Pickering stabilizer dispersion liquid and 0% -2.0% of protective agent.
  2. 2. The salmonella phage nanoemulsion formulation of claim 1, wherein the salmonella phage suspension is prepared by dissolving and diluting salmonella phage with PBS buffer solution after activating salmonella phage, and the titer of phage in the salmonella phage suspension is 10 6 ～10 9 PFU/mL.
  3. 3. The salmonella phage nanoemulsion formulation of claim 1, wherein the edible oil is one or a mixture of more than two of soybean oil, corn oil and medium chain triglycerides, the Pickering stabilizer is one or a mixture of more than two of nanocellulose, modified starch particles and protein-polysaccharide composite particles, and the protectant is one or a mixture of more than two of gelatin, whey protein, trehalose and sucrose.
  4. 4. The salmonella phage nanoemulsion formulation of claim 1, wherein the average particle size of the salmonella phage nanoemulsion formulation is 100-230 nm.
  5. 5. The method for preparing the salmonella phage nanoemulsion formulation of any one of claims 1-4, comprising the steps of: (1) After activating salmonella phage, dissolving and diluting the salmonella phage with PBS buffer solution to prepare salmonella phage suspension; (2) Adding Pickering stabilizer into water phase buffer solution, and dispersing under high-speed stirring to form uniform particle Pickering stabilizer dispersion; (3) Adding edible oil into the Pickering stabilizer dispersion liquid obtained in the step (2) in a continuous or fractional adding mode, and pre-emulsifying under a high-speed shearing condition to obtain coarse emulsion; (4) Homogenizing or ultrasonic treating the crude emulsion obtained in the step (3) under high pressure to obtain nanoemulsion; (5) And (3) adding the salmonella phage suspension obtained in the step (1) into the nanoemulsion obtained in the step (4), uniformly mixing under a low shearing condition, adding a protective agent, stirring, and then adjusting the pH to 6.4-6.8 to obtain the salmonella phage nanoemulsion preparation.
  6. 6. The preparation method of the salmonella phage nanoemulsion preparation of claim 5, wherein the aqueous phase buffer solution in the step (2) is food grade phosphate buffer solution PBS, the high-speed stirring condition is that the aqueous phase buffer solution is dispersed for 10-20 min under the high-speed dispersion of 3000-12000 rpm, and the mass percentage of the Pickering stabilizer in the Pickering stabilizer dispersion solution is 0.5-3.0%.
  7. 7. The method for preparing a salmonella phage nanoemulsion formulation according to claim 5, wherein the high-speed shearing condition in step (3) is shearing at 5000-20000 rpm for 10-15 min.
  8. 8. The preparation method of the salmonella phage nanoemulsion preparation of claim 5, wherein the high-pressure homogenization pressure in the step (4) is 10-80 MPa, the homogenization times are 5-8, the power of ultrasonic treatment is 200-1200W, the treatment time is 15-20 min, and the temperature in the high-pressure homogenization or ultrasonic treatment process is not higher than 25 ℃.
  9. 9. The method for preparing a salmonella phage nanoemulsion formulation according to claim 5, wherein the rotation speed of the low shear condition in step (5) is 100-800 rpm, and the time is 10-30 min.
  10. 10. Use of the salmonella phage nanoemulsion formulation of any one of claims 1-4 in bacteriostatic preservation of meat products.

Description

Salmonella phage nanoemulsion preparation and preparation method and application thereof Technical Field The invention belongs to the technical field of food preservation and antibacterial delivery, and particularly relates to a salmonella phage nanoemulsion preparation, a preparation method and application thereof. Background Meat products are susceptible to microbial contamination during processing, transportation and storage. Phage has the advantages of strong targeting, low residual risk and the like as a biological antibacterial factor, but the phage is possibly influenced by factors such as interfacial shear, oxidation, storage layering and the like in an emulsion system to reduce the activity. The existing phage emulsion system mostly adopts surfactants such as Tween/span/phospholipid and the like and vegetable oil phase to construct emulsion drop structure, and although the dispersibility can be improved, the problems of insufficient interfacial film strength, easiness in aggregation and layering after long-term storage, activity maintenance fluctuation and the like still exist. Accordingly, there is a need to provide an emulsion carrier system that is more stable in food systems, is capable of improving phage activity retention, and is more suitable for surface coating of meat products. Disclosure of Invention The invention aims to provide a salmonella phage nanoemulsion preparation. The invention also aims at providing a preparation method of the salmonella phage nanoemulsion preparation. The invention also provides an application of the salmonella phage nanoemulsion preparation in bacteriostasis and fresh-keeping of meat products. The first aim of the invention can be achieved by adopting the following technical scheme that the salmonella phage nanoemulsion preparation mainly comprises, by mass, 5% -15% of salmonella phage suspension, 84% -94% of edible oil, 0.5% -3.0% of Pickering stabilizer dispersion liquid and 0% -2.0% of protective agent. In the salmonella phage nanoemulsion formulation of the present invention: Preferably, the salmonella phage suspension is prepared by dissolving and diluting salmonella phage with PBS buffer solution after activating the salmonella phage, and the titer of phage in the salmonella phage suspension is 10 6～109 PFU/mL. The Salmonella phage (Salmonella) of the present invention can be any publicly available Salmonella phage. Preferably, the salmonella phage of the present invention recommends, but is not limited to, salmonella phage with a preservation number of cctccc No. M2021765. Preferably, the PBS buffer solution is food-grade Phosphate Buffer Solution (PBS), and the PBS buffer solution comprises sodium chloride, disodium hydrogen phosphate, monopotassium phosphate and the like, and meets the requirements of GB 2760 or FDA food contact materials. Preferably, the edible oil of the present invention includes, but is not limited to, one or a mixture of two or more of soybean oil, corn oil and medium chain triglycerides. Preferably, the Pickering stabilizer is one or a mixture of more than two of nanocellulose, modified starch particles and protein-polysaccharide composite particles. The nano-cellulose refers to food-grade Cellulose Nanocrystals (CNC) or microfibrillated cellulose (MFC), the modified starch particles refer to common modified starch such as food-grade sodium starch octenyl succinate, acetylated distarch phosphate and the like, and the protein-polysaccharide composite particles refer to edible composite particles formed by known methods such as electrostatic compositing and heat treatment of food-grade proteins (such as whey proteins and soy protein isolates) and food-grade polysaccharides (such as gum and acacia). The Pickering stabilizer is edible solid particles, and the solid particles can be adsorbed on an oil-water interface to form a particle film so as to stabilize nano emulsion drops. Preferably, the protective agent is one or a mixture of more than two of gelatin, whey protein, trehalose and sucrose. Further, the mass percentage of the protective agent is 0.01% -2.0%, and more preferably 0.5% -1.5%. Preferably, the average particle size of the salmonella phage nanoemulsion preparation is 100-230 nm. The edible oil, the Pickering stabilizer, the protective agent and the PBS buffer solution all refer to food-grade or food additive-grade products meeting the requirements of relevant regulations such as national food safety standards in China or GB 2760 food additive use standards. These materials are commercially available as known in the art and are available from conventional commercial sources. The salmonella phage nanoemulsion preparation does not contain tween-type surfactant or span-type surfactant, so that emulsion drop stability is realized by using a solid particle interface film. The salmonella phage nanoemulsion preparation is a solid particle stable (Pickering) salmonella phage nanoemulsion preparation, a stable particle film i