In recent years, the definition and role of food have undergone significant changes. Beyond traditional nutritional and sensory values, the health benefits and disease prevention potential of food have gained increasing attention. Food safety has become a major challenge in modern society, particularly due to the risk of contamination by pathogenic bacteria during food storage and transportation. Common foodborne pathogens such as Salmonella, Escherichia coli, and Listeria can cause food poisoning, gastroenteritis, and even more severe health issues upon ingestion. According to the World Health Organization (WHO) reports, approximately 600 million people worldwide are affected by foodborne illnesses annually, and the growing antimicrobial resistance of pathogens poses new challenges to traditional chemical preservatives [1-2]. Therefore, developing green and safe food preservation technologies is a critical pathway to promote the development of “green food,” meeting the growing consumer demand for safe and healthy food.

Currently, mature food preservation methods include chemical preservatives, physical techniques, and biological preservatives. Common chemical preservatives, such as benzoic acid, sorbic acid, nitrites, and propionates, extend food shelf life by inhibiting microbial growth or killing bacteria. While cost-effective and easy to use, long-term consumption of these chemicals may trigger allergic reactions, immune system damage, and even contribute to chronic diseases like cancer. Additionally, some pathogens may develop resistance due to genetic mutations, reducing the efficacy of preservatives and increasing food safety risks [2]. Physical methods, such as high-temperature sterilization (e.g., pasteurization), refrigeration, irradiation, and vacuum packaging, are safer as they avoid chemical additives. However, these methods require specialized equipment, are costly, and may not be suitable for foods sensitive to high temperatures. Biological preservatives, such as lactic acid bacteria, bacteriophages, and bacteriocins (e.g., nisin), inhibit pathogens by producing natural antimicrobial substances. These are relatively safe and non-toxic but have a narrow antimicrobial spectrum and may be less effective in certain food matrices [3].

With growing environmental awareness and the promotion of sustainable development, green and eco-friendly food preservation technologies have become a trend in industry. Natural antimicrobial agents, such as lysozyme and antimicrobial peptides (AMPs), have low toxicity, degrade into harmless substances, and align with sustainability and environmental goals. Their use can replace chemical preservatives, reduce environmental pollution and toxicity, and promote the green transformation of food preservation technologies [4]. Unlike chemical preservatives, which often compromise food sensory qualities (e.g., color, taste, and odor), lysozyme and AMPs are highly specific, inhibiting harmful bacteria while preserving food flavor and nutritional content. Lysozyme degrades bacterial cell walls, while AMPs disrupt bacterial membranes, minimally affecting food quality [4-5]. Thus, developing natural preservation technologies not only extends shelf life but also maintains food nutrition and sensory attributes, enhancing consumer satisfaction and supporting the demand for high-quality food.

Chemical food preservatives, widely used to extend shelf life, have increasingly come under scrutiny due to documented associations with carcinogenicity, hypersensitivity reactions, and metabolic disorders. These concerns have led to rising public and scientific interest in replacing chemical preservatives with safer, biologically derived alternatives.

Natural antimicrobial agents, particularly lysozyme and antimicrobial peptides (AMPs), may play a key role in food preservation. Lysozyme, a peptidoglycan-N-acetylmuramoyl hydrolase commonly derived from hen egg white, has been evaluated by the European Food Safety Authority as safe even at levels far exceeding those typically used in food. AMPs, such as defensins, exert potent antimicrobial effects through mechanisms like membrane permeabilization and inhibition of cell wall synthesis.

Compared to chemical preservatives, AMPs and lysozymes are biodegradable and can target a wide range of foodborne pathogens (Figure. 1). However, natural extraction from animal or plant sources can be costly and inefficient. By leveraging synthetic biology, these antimicrobial proteins can be produced economically and at scale using microbial expression systems.

Figure 1. The comparison between antibiotics and antimicrobial piptides.

Food waste and human health problems caused by food corruption are more and more widespread, so it is urgent to study a safe and effective preservative. Bacterial growth is an important cause of food spoilage, so whether it can inhibit the growth of bacteria is an important criterion to measure the effectiveness of preservatives. At present, the research tends to find safer natural antibacterial substances for food preservation, among which antibacterial peptides and lysozyme are two important natural antibacterial products, both of which can inhibit bacterial growth in a broad spectrum. In this topic, we choose lysozyme from human sources((hLYZ, GenBank ID: NP_000230)) for the production of biological preservatives, mainly because lysozyme from other sources may cause allergies to human bodies and reduce the side effects of food preservatives. Compared with lysozyme, the reported antimicrobial peptides are richer and more diverse. Therefore, in order to find effective and safe antimicrobial peptides, we found 74 possible antimicrobial peptide molecules by consulting the literature, and eliminated the antimicrobial peptides with hemolysis risk and long intestinal half-life by online tools (HemoPI2 and ToxinPred), and finally screened out 9 possible candidate antimicrobial peptide molecules.

⦁ Candidate Selection:

We employed a comprehensive in silico screening pipeline to identify AMPs with low predicted hemolytic activity, low predicted toxicity and acceptable intestinal stability.

Table1. List of AMP activity analysis tools

Name	Activity	Link
HemoPI/Hemolytik	Predicts hemolytic and non-hemolytic peptides	https://webs.iiitd.edu.in/raghava/hemopi/index.php
ToxinPred	Predicts toxic and non-toxic peptides	https://webs.iiitd.edu.in/raghava/toxinpred/
HLP	Peptide half-life prediction in intestinal environment	http://crdd.osdd.net/raghava/hlp/pep_both.htm

Table 2. Structure and characteristics of AMPs

candidates were selected: Cecropin A, Melittin, Buforin II, LL-37, Plectasin, Hepcidin, Piscidin-1, and Lactoferricin. We also design a antisepsis mix protein, which connecting lysosome and 9 AMPs by flexible linker (GGGGS).

Based on the study of food preservation with lysozyme and antimicrobial peptides, our preliminary results show that the combination of lysozyme and antimicrobial peptides plays an important role in inhibiting bacterial growth. Therefore, we can use the combination of antimicrobial peptides and lysozyme as natural preservatives for food preservation. Specifically, our products can be added to food in the form of concentrated solution, for example, in all aspects of meat products, dairy products or fruit and vegetable processing, our liquid preservatives can be coated or soaked on the surface of food to prevent the growth of bacteria, so as to achieve the purpose of food preservation and antisepsis.

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