OBJECTIVE: This study evaluates the antibacterial characteristics and mechanisms of combined tea polyphenols (TPs), Nisin, and ε-polylysine (PL) against Streptococcus canis, Streptococcus minor, Streptococcus mutans, and Actinomyces oris, common zoonotic pathogens in companion animals.
RESULTS: Pathogenic strains were isolated from feline oral cavities and assessed using minimum inhibitory concentration (MIC) tests, inhibition zone assays, growth kinetics, and biofilm inhibition studies. Among single agents, PL exhibited the lowest MIC values against all four pathogens. TP showed significant resistance against S. minor, and Nisin against S. mutans. The combination treatment (Comb) of TP, Nisin, and PL in a ratio of 13:5:1 demonstrated broad-spectrum antibacterial activity, maintaining low MIC values, forming large inhibition zones, prolonging the bacterial lag phase, reducing growth rates, and inhibiting biofilm formation. RNA sequencing and metabolomic analysis indicated that TP, Nisin, and PL inhibited various membrane-bound carbohydrate-specific transferases through the phosphoenolpyruvate-dependent phosphotransferase system in S. canis, disrupting carbohydrate uptake. They also downregulated glycolysis and the citric acid cycle, inhibiting cellular energy metabolism. Additionally, they modulated the activities of peptidoglycan glycosyltransferases and d-alanyl-d-alanine carboxypeptidase, interfering with peptidoglycan cross-linking and bacterial cell wall stability.
CONCLUSIONS: The Comb therapy significantly enhances antibacterial efficacy by targeting multiple bacterial pathways, offering potential applications in food and pharmaceutical antimicrobials.

Widespread outbreaks of threatening infections caused by unknown pathogens and water transmission have spawned the development of adsorption methods for pathogen elimination. We proposed a biochar functionalization strategy involving ε-polylysine (PLL), a bio-macromolecular poly(amino acid)s with variable folding conformations, as a \"pathogen gripper\" on biochar. PLL was successfully bridged onto biochar via polydopamine (PDA) crosslinking. The extension of electropositive side chains within PLL enables the capture of both nanoscale viruses and micrometer-scale bacteria in water, achieving excellent removal performances. This functionalized biochar was tentatively incorporated into ultrafiltration (UF) system, to achieve effective and controllable adsorption and retention of pathogens, and to realize the transfer of pathogens from membrane surface/pore to biochar surface as well as flushing water. The biochar-amended UF systems presents complete retention (∼7 LRV) and hydraulic elution of pathogens into membrane flushing water. Improvements in removal of organics and anti-fouling capability were observed, indicating the broken trade-off in UF pathogen removal dependent on irreversible fouling. Chemical characterizations revealed adsorption mechanisms encompassing electrostatic/hydrophobic interactions, pore filling, electron transfer, chemical bonding and secondary structure transitions. Microscopic and mechanical analyses validated the mechanisms for rapid adsorption and pathogen lysis. Low-concentration alkaline solution for used biochar regeneration, facilitated the deprotonation and transformation of PLL side chain to folded structures (α-helix/β-sheet). Biochar regeneration process also promoted the effective detachment/inactivation of pathogens and protection of functional groups on biochar, corroborated by physicochemical inspection and molecular dynamics simulation. The foldability of poly(amino acid)s acting like dynamic arms, significantly contributed to pathogen capture/desorption/inactivation and biochar regeneration. This study also inspires future investigation for performances of UF systems amended by poly(amino acid)s-functionalized biochar under diverse pressure, temperature, reactive oxygen species of feeds and chemical cleaning solutions, with far-reaching implications for public health, environmental applications of biochar, and UF process improvement.

In light of the commendable advantages inherent in natural polymers such as biocompatibility, biodegradability, and cost-effectiveness, researchers are actively engaged in the development of biopolymer-based biodegradable food packaging films (BFPF). However, a notable limitation is that most biopolymers lack intrinsic antimicrobial activity, thereby restricting their efficacy in food preservation. To address this challenge, various active substances with antibacterial properties have been explored as additives to BFPF. Among these, ε-polylysine has garnered significant attention in BFPF applications owing to its outstanding antibacterial properties. This study provides a brief overview of the synthesis method and chemical properties of ε-polylysine, and comprehensively examines its impact as an additive on the properties of BFPF derived from diverse biopolymers, including polysaccharides, proteins, aliphatic polyesters, etc. Furthermore, the practical applications of various BFPF functionalized with ε-polylysine in different food preservation scenarios are summarized. The findings underscore that ε-polylysine, functioning as an antibacterial agent, not only directly enhances the antimicrobial activity of BFPF but also serves as a cross-linking agent, interacting with biopolymer molecules to influence the physical and mechanical properties of BFPF, thereby enhancing their efficacy in food preservation.

Traditional petroleum-based food-packaging materials have poor permeability, limited active packaging properties, and difficulty in biodegradation, limiting their application. We developed a carboxymethylated tamarind seed polysaccharide composite film incorporated with ε-polylysine (CTPε) for better application in fresh-cut agricultural products. The CTPε films exhibit excellent water vapor barrier properties, but the mechanical properties are slightly reduced. Fourier transform infrared spectroscopy and X-ray diffraction spectra indicate the formation of hydrogen bonds between ε-PL and CTP, leading to their internal reorganization and dense network structure. With the increase of ε-PL concentration, composite films showed notable inhibition of postharvest pathogenic fungi and bacteria, a significant enhancement of 2,2\'- azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical-scavenging activity, and gradual improvement of wettability performance. Cytotoxicity experiments confirmed the favorable biocompatibility when ε-PL was added at 0.3% (CTPε2). In fresh-cut bell pepper preservation experiments, the CTPε2 coating effectively delayed weight loss and malondialdehyde increase preserved the hardness, color, and nutrients of fresh-cut peppers and prolonged the shelf life of the fresh-cut peppers, as compared with the control group. Therefore, CTPε composite films are expected to be a valuable packaging material for extending the shelf life of freshly cut agricultural products.

Biocomplex materials formed by oppositely charged biopolymers (proteins) tend to be sensitive to environmental conditions and may lose part functional properties of original proteins, and one of the approaches to address these weaknesses is protein modification. This study established an electrostatic composite system using succinylated ovalbumin (SOVA) and ε-polylysine (ε-PL) and investigated the impact of varying degrees of succinylation and ε-PL addition on microstructure, environmental responsiveness and functional properties. Molecular docking illustrated that the most favorable binding conformation was that ε-PL binds to OVA groove, which was contributed by the multi‑hydrogen bonding and hydrophobic interactions. Transmission electron microscopy observed that SOVA/ε-PL had a compact spherical structure with 100 nm. High-degree succinylation reduced complex sensitivity to heat, ionic strength, and pH changes. ε-PL improved the gel strength and antibacterial properties of SOVA. The study suggests possible uses of SOVA/ε-PL complex as multifunctional protein complex systems in the field of food additives.

This study investigated the effects of nisin combined with ε-polylysine on microorganisms and the refrigerated quality of fresh-cut jackfruit. After being treated with distilled water (control), nisin (0.5 g/L), ε-polylysine (0.5 g/L), and the combination of nisin (0.1 g/L) and ε-polylysine (0.4 g/L), microporous modified atmosphere packaging (MMAP) was carried out and stored at 10 ± 1°C for 8 days. The microorganisms and physicochemical indexes were measured every 2 days during storage. The results indicated that combined treatment (0.1 g/L nisin, 0.4 g/L ε-polylysine) had the best preservation on fresh-cut jackfruit. Compared with the control, combined treatment inhibited microbial growth (total bacterial count, mold and yeast), reduced the weight loss rate, respiratory intensity, polyphenol oxidase and peroxidase activities, and maintained higher sugar acid content, firmness, and color. Furthermore, it preserved higher levels of antioxidant compounds, reduced the accumulation of malondialdehyde and hydrogen peroxide, thereby reducing oxidative damage and maintaining high nutritional and sensory qualities. As a safe application of natural preservatives, nisin combined with ε-polylysine treatment has great application potential in the fresh-cut jackfruit industry.

Carvacrol is well-known natural antimicrobial compounds. However, its usage in fruit preservation is restricted owing to poor water solubility. Our study aims to address this limitation by combining carvacrol with whey protein isolate (WPI) to form nanoemulsion and enhancing antimicrobial properties and stability of nanoemulsion through ε-polylysine addition, thereby improving their application in fruit preservation. The results indicated that the nanoemulsion exhibited a double-layer structure. The physicochemical properties and storage stability were found to be favorable under the conditions of WPI (0.3 wt% v/v), Carvacrol (0.5 % v/v), and ε-polylysine (0.3 wt% v/v). In addition, the nanoemulsion had inhibitory effects on Staphylococcus aureus, Escherichia coli, and Aspergillus niger at concentrations of minimal inhibition concentration (32, 32, and 200 μg/mL, respectively). In addition, during a 7-day storage period, the nanoemulsion effectively preserved mangoes. Therefore, nanoemulsion could serve as a candidate for control of postharvest mangoes spoilage and extend its period of storage.

The aim of this investigation was to scrutinize the effects of a thermal treatment on the electrostatic complex formed between gum arabic (GA) and ε-polylysine (ε-PL), with the goal of improving the antibacterial properties and reducing the hygroscopicity of ε-PL. The heated complex with a ratio of 1:4 exhibited an encapsulation efficiency of 93.3%. Additionally, it had an average particle size of 350.3 nm, a polydispersity index of 0.255, and a zeta potential of 18.9 mV. The formation of the electrostatic complex between GA and ε-PL was confirmed through multispectral analysis, which demonstrated the participation of hydrogen bonding and hydrophobic and electrostatic interactions, as well as the enhanced effect of heat treatment on these forces within the complex. The complex displayed a core-shell structure, with a regular distribution and a shape that was approximately spherical, as observed in the transmission electron microscopy images. Additionally, the heated GA-ε-PL electrostatic composite exhibited favorable antibacterial effects on Salmonella enterica and Listeria monocytogenes, with reduced minimum inhibitory concentrations (15.6 μg/mL and 62.5 μg/mL, respectively) and minimum bactericidal concentrations (31.3 μg/mL and 156.3 μg/mL, respectively) compared to free ε-PL or the unheated electrostatic composite. Moreover, the moisture absorption of ε-PL reduced from 92.6% to 15.0% in just 48 h after being incorporated with GA and subsequently subjected to heat. This research showed a way to improve the antibacterial efficiency and antihygroscopicity of ε-PL, reducing its application limitations as an antimicrobial substance to some extent.

This study provides a novel strategy for preparing bio-based antibacterial emulsions stabilized by cellulose nanocrystals (CNCs). Antibacterial ε-polylysine (ε-PL) with a positive charge was introduced into the aqueous phase to modulate the interfacial behavior of CNCs via electrostatic interactions. Pickering emulsions containing ε-PL/CNCs (ε-PL 0.07-0.1 g/L) had significantly better stability, larger emulsion ratio, smaller emulsion droplet diameter, and superior antibacterial ability than emulsions stabilized by CNCs alone. This could be attributed to the formation of a CNC-dense layer at the interface in the continuous phase caused by a reduction of electrostatic repulsion after adding ε-PL. This was confirmed by zeta potential measurements, rheological properties, and bio-freezing scanning electron microscopy. In addition, cinnamaldehyde was introduced into the oil phase to further improve the antibacterial properties of the emulsion, thereby avoiding easy evaporation into water. Our findings provide an innovative solution for preparing bio-based antibacterial emulsions stabilized by ε-PL/CNCs, which will benefit the development of food, medicine, and cosmetic lotions.

The shelf life of beef is shortened by microbial infection, which limits its supply in the market. Active packaging film is expected to overcome this difficulty. In this study, an antibacterial/antioxidant SS-ε-PL-TA biocomposite film made by soy protein isolate/sodium alginate/ε-polylysine/tannic acid was designed and prepared. Due to the formation of hydrogen bonds and enhanced hydrophobic interactions, the biocomposite film showed enhanced mechanical property. Tensile strength increased from 22.8 ± 2.59 MPa to 64.34 ± 6.22 MPa, and elongation at break increased from 7.70 ± 1.07 % to 13.98 ± 0.22 %. The composite film displayed excellent antibacterial activity owing to the damage to cell membranes and biofilms of bacteria. Furthermore, the antioxidant activity also significantly increased (DPPH ∙ scavenging activity was 78.0 %). The shelf life of beef covered with the SS-ε-PL-TA film was extended by 3 days compared to the control group by decreasing lipid oxidation and inhibiting bacterial growth, showing a good application potential in food packaging.