An edible bilayer film incorporated with lysozyme based on chitosan and sodium alginate was prepared via the layer‐by‐layer method. The film was characterized and its effects against the fish spoilage bacteria Pseudomonas fluorescens and Shewanella putrefaciens were investigated and compared to the monolayer films. The results suggest that electrostatic interactions such as hydrogen bonds occurred between the two layers of the bilayer film. Compared with monolayer films, the mechanical and gas barrier properties of the bilayer film were improved and higher inhibitory activity against both bacteria was exhibited. After treatment with the bilayer film, scanning electron microscopy revealed the serious damage to the bacterial cell surface, and cell membrane permeability and nucleic acid leakage increased. The bilayer film also affected the activity of alkaline phosphatase and adenosine triphosphatase. SDS‐PAGE of bacterial total protein revealed that the protein concentration n decreased, which may be due to the leakage of proteins from the damaged bacterial cell. All the results indicate that the bilayer film possesses good physicochemical properties and can destroy the bacterial cell membrane. Such superior antibacterial properties against fish spoilage bacteria indicate a good potential application of bilayer films in fish preservation.

中文翻译：

---

掺有溶菌酶的双层可食膜的制备及其对鱼腐败细菌的影响

用溶菌酶基于脱乙酰壳多糖和藻酸钠掺入的可食用双层膜制备通过在层-层的方法。这部电影的特点是，它对鱼类腐败菌荧光假单胞菌和腐烂希瓦氏菌的影响被研究并与单层膜比较。结果表明在双层膜的两层之间发生了诸如氢键之类的静电相互作用。与单层膜相比，双层膜的机械和气体阻隔性能得到改善，并且表现出对两种细菌更高的抑制活性。用双层膜处理后，扫描电子显微镜显示出对细菌细胞表面的严重破坏，并且细胞膜通透性和核酸泄漏增加。双层膜还影响碱性磷酸酶和腺苷三磷酸酶的活性。对细菌总蛋白的SDS-PAGE分析表明，蛋白浓度n降低，这可能是由于蛋白从受损细菌细胞中泄漏所致。所有结果表明该双层膜具有良好的理化性质并且可以破坏细菌细胞膜。这种对鱼类变质细菌的优异抗菌性能表明双层膜在鱼类保鲜中具有良好的潜在应用前景。