细菌由于其独特的特性，如基因可操纵性、快速增殖和对目标疾病部位的特异性，已被广泛用作疾病治疗的活疗法。然而，各种体内损伤会降低给药细菌的活力，导致总体生物利用度较低。此外，细菌表面的固有抗原以及释放的毒素和代谢物可能会导致不良的安全问题。这些限制不可避免地导致治疗结果不足，从而阻碍活细菌疗法的临床转化。最近，我们开发了一个多功能平台，通过化学修饰或物理封装单独纳米涂层细菌来制备先进的活细菌疗法，这可以提高生物利用度并减少增强微生物疗法的副作用。在这里，我们使用界面自组装来制备脂膜包被的细菌（LCB），由于纳米涂层的保护能力，该细菌对各种恶劣的环境条件表现出增强的抵抗力。同时，我们应用机械挤压来生成细胞膜包被细菌（CMCB），由于纳米涂层的屏蔽作用，其生物相容性得到了改善。我们描述了它们的详细制备过程并展示了包被细菌的预期功能。我们还表明，在小鼠模型中口服给药和静脉注射后，LCB 和 CMCB 分别在治疗结肠炎和肿瘤方面表现出诱人的潜力。与缺乏通用的异质表达分子工具的生物工程相比，表面纳米涂层技术可以方便地引入功能成分，且不受菌株类型的限制。不包括细菌培养，LCB 的制造需要约 2 小时，而 CMCB 的制备需要约 5 小时。© 2024。Springer Nature Limited。

Bacteria have been extensively utilized as living therapeutics for disease treatment due to their unique characteristics, such as genetic manipulability, rapid proliferation and specificity to target disease sites. Various in vivo insults can, however, decrease the vitality of dosed bacteria, leading to low overall bioavailability. Additionally, the innate antigens on the bacterial surface and the released toxins and metabolites may cause undesired safety issues. These limitations inevitably result in inadequate treatment outcomes, thereby hindering the clinical transformation of living bacterial therapeutics. Recently, we have developed a versatile platform to prepare advanced living bacterial therapeutics by nanocoating bacteria individually via either chemical decoration or physical encapsulation, which can improve bioavailability and reduce side effects for enhanced microbial therapy. Here we use interfacial self-assembly to prepare lipid membrane-coated bacteria (LCB), exhibiting increased resistance against a variety of harsh environmental conditions owing to the nanocoating's protective capability. Meanwhile, we apply mechanical extrusion to generate cell membrane-coated bacteria (CMCB), displaying improved biocompatibility owing to the nanocoating's shielding effect. We describe their detailed preparation procedures and demonstrate the expected functions of the coated bacteria. We also show that following oral delivery and intravenous injection in mouse models, LCB and CMCB present appealing potential for treating colitis and tumors, respectively. Compared with bioengineering that lacks versatile molecular tools for heterogeneous expression, the surface nanocoating technique is convenient to introduce functional components without restriction on bacterial strain types. Excluding bacterial culture, the fabrication of LCB takes ~2 h, while the preparation of CMCB takes ~5 h.© 2024. Springer Nature Limited.