KR-102962817-B1 - Bacteriophage stabilization composition and sterilization method using the same

Abstract

The present invention relates to a composition for stabilizing bacteriophages, a bacteriophage composition containing the same, a food packaging material, and a sterilization method using the same. Compared to conventional bacteriophage storage methods, the bacteriophage stabilization composition according to the present invention enables commercialization and storage in a more convenient, economical, and efficient manner even at room temperature. Furthermore, by manufacturing a bacteriophage composition containing the bacteriophage stabilization composition or packaging materials coated with bacteriophages, it has the effect of being directly applied to food for biological control, and thus can be usefully employed for preventing bacterial contamination of food and maintaining food safety.

Inventors

• 임정아
• 김어진
• 우민아
• 임민철

Assignees

• 한국식품연구원

Dates

Publication Date

20260511

Application Date

20230602

Claims (15)

1. A composition for stabilizing bacteriophages comprising sorbitol, sodium chloride, and magnesium sulfate, A composition for stabilizing bacteriophages, wherein the above-mentioned sorbitol is included at a concentration of 10 to 15% (w/v).
2. In paragraph 1, A composition for stabilizing a bacteriophage, wherein the bacteriophage has a specific killing ability against Escherichia coli , Staphylococcus aureus , Salmonella , Bacillus cereus , Listeria monocytogenes , Vibrio parahaemolyticus , or Cronobacter sakazakii .
3. delete
4. A bacteriophage composition comprising a bacteriophage stabilization composition according to claim 1.
5. In paragraph 4, A bacteriophage composition having a specific killing ability against Escherichia coli , Staphylococcus aureus , Salmonella , Bacillus cereus , Listeria monocytogenes , Vibrio parahaemolyticus , or Cronobacter sakazakii .
6. In paragraph 4, The bacteriophage composition comprises the bacteriophage in an amount of 10⁴ to 10¹² PFU/ml.
7. A coating composition comprising a bacteriophage stabilization composition according to claim 1, a bacteriophage, and a coating material.
8. In Paragraph 7, A coating composition in which the above-mentioned bacteriophage has a specific killing ability against Escherichia coli , Staphylococcus aureus , Salmonella , Bacillus cereus , Listeria monocytogenes , Vibrio parahaemolyticus , or Cronobacter sakazakii .
9. In Paragraph 7, The coating composition, wherein the coating material is pullulan, carboxymethyl cellulose, or polyvinyl alcohol.
10. In Paragraph 9, A coating composition in which the above coating material is pullulan or carboxymethyl cellulose.
11. Antimicrobial film manufactured using a coating composition according to claim 7.
12. Food packaging material comprising the antimicrobial film of claim 11.
13. A method for sterilizing food or food ingredients, comprising the step of treating the food or food ingredients with a bacteriophage composition according to paragraph 4.
14. A method for sterilizing food or food ingredients, comprising the steps of: placing food or food ingredients into a container packaged with a food packaging material according to claim 12; and bringing said container into contact with water.
15. In paragraph 13 or 14, The above method is a sterilization method for sterilizing Escherichia coli , Staphylococcus aureus , Salmonella , Bacillus cereus , Listeria monocytogenes , Vibrio parahaemolyticus , or Cronobacter sakazakii .

Description

Bacteriophage stabilization composition and sterilization method using the same The present invention relates to a composition for stabilizing bacteriophages, a bacteriophage composition containing the same, a food packaging material, and a sterilization method using the same. Food is highly likely to be contaminated by pathogens during manufacturing, distribution, and storage processes. Since contamination by pathogens not only degrades quality but can also cause food poisoning upon consumption, it is important to prevent food contamination by pathogens. However, despite significant advancements in food hygiene technology, various foodborne pathogens continue to threaten human health, leading to hospitalization and death (Elbehiry, et al., 2023). Among various foods, fresh foods are cited as a major cause of food poisoning, harboring virulent pathogens such as Escherichia coli , Salmonella , and Listeria monocytogenes . In particular, E. coli threatens the health of millions of people every year, and outbreaks of E. coli O157:H7 infection occur very frequently with the consumption of contaminated plant-based foods (e.g., lettuce, spinach, tomatoes, and fresh fruits) (Puligundla & Lim, 2022). This is because fruits and vegetables are grown in open environments vulnerable to microbial contamination and are often not treated to reduce or eliminate pathogens. Since agricultural products are foods that can be eaten immediately without undergoing a separate sterilization process, it is important to remove microbial load during pre-treatment steps such as washing or post-treatment steps such as using edible disinfectants. Accordingly, various methods have been developed and devised to enhance the safety of fresh foods. Natural or chemical food preservatives are used to control foodborne illnesses. Natural preservatives such as organic acids, bacteriocins, chitosan, and lactoferrin have weak antimicrobial activity and limited use, while chemical preservatives and antibiotics are more effective but raise concerns regarding side effects. For example, chlorine-based disinfectants are widely used to eliminate microorganisms contaminating fresh foods due to their high antimicrobial efficacy; however, there are efforts to reduce their use due to concerns about the generation of carcinogenic byproducts that pose risks to the environment and health. To address these issues, many researchers are developing alternative disinfectants. Bacteriophages are gaining attention as effective antibiotics as they complement the limitations of previous methods. Bacteriophages are viruses that infect and kill bacteria; they specifically bind to target bacteria, reproduce within them, and lyse the bacteria. Due to their high specificity, phages do not affect the normal and beneficial microbial communities of the host, food, or environment. Furthermore, since they do not have harmful effects on food quality or animal and human health, additional removal steps are not required. Because of these advantages, bacteriophages can be applied directly to food to control foodborne pathogens. As safe biological materials recognized as generally recognized as safe (GRAS) by the U.S. Food and Drug Administration (FDA) since 2006, bacteriophages are being applied as food additives to prevent food contamination by pathogens. Since the approval of the first bacteriophage formulation, additional bacteriophage products such as Salmo Fresh™ for Salmonella control and EcoShield® for E. coli control have been approved and are currently on the market. Meanwhile, for the development of these phage products and their application in food, storage methods and shelf life are critical. The commercialization and preservation of phages can be achieved using low-temperature storage of aqueous suspensions, freeze-drying, encapsulation, and spray-drying. Since the titer of phages decreases rapidly when stored at room temperature, the primary strategy used for preservation is storage at low temperatures, such as 4°C, -20°C, and -80°C. In practice, bacteriophage products such as Salmo Fresh™ for Salmonella control and EcoShield® for E. coli control are distributed in solution form, with instructions to store containers under refrigerated conditions (2–8°C) and keep them shielded from light. The main disadvantages of these storage methods are their high cost and unsuitability for large-scale storage. Another method is encapsulation, which involves wrapping and protecting the phage with biomaterial. Encapsulated phages exhibit greater resistance to environmental changes during storage and shipping. However, encapsulation technology has not improved the viability of bacteriophages compared to low-temperature storage (including refrigeration), and additional costs may be incurred as new storage methods must be developed depending on the application. Lyophilization is an effective preservation method that maintains structural integrity and extends shelf life by sublimating wat