SummaryPlants offer a promising chassis for the large‐scale, cost‐effective production of diverse therapeutics, including antimicrobial peptides (AMPs). However, key advances will reduce production costs, including simplifying the downstream processing and purification steps. Here, using Nicotiana benthamiana plants, we present an improved modular design that enables AMPs to be secreted via the endomembrane system and sequestered in an extracellular compartment, the apoplast. Additionally, we translationally fused an AMP to a mutated small ubiquitin‐like modifier sequence, thereby enhancing peptide yield and solubilizing the peptide with minimal aggregation and reduced occurrence of necrotic lesions in the plant. This strategy resulted in substantial peptide accumulation, reaching around 2.9 mg AMP per 20 g fresh weight of leaf tissue. Furthermore, the purified AMP demonstrated low collateral toxicity in primary human skin cells, killed pathogenic bacteria by permeabilizing the membrane and exhibited anti‐infective efficacy in a preclinical mouse (Mus musculus) model system, reducing bacterial loads by up to three orders of magnitude. A base‐case techno‐economic analysis demonstrated the economic advantages and scalability of our plant‐based platform. We envision that our work can establish plants as efficient bioreactors for producing preclinical‐grade AMPs at a commercial scale, with the potential for clinical applications.

中文翻译：

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在小鼠模型中以植物为基础高产地生产具有有效活性的抗菌肽


摘要植物为大规模、经济有效地生产包括抗菌肽（AMP）在内的多种治疗药物提供了一个有前途的基础。然而，关键的进步将降低生产成本，包括简化下游加工和纯化步骤。在这里，我们利用本塞姆氏烟草植物，提出了一种改进的模块化设计，使 AMP 能够通过内膜系统分泌并隔离在细胞外室（质外体）中。此外，我们将 AMP 翻译融合到突变的小泛素样修饰序列上，从而提高肽产量并以最小的聚集溶解肽，并减少植物中坏死病变的发生。该策略导致大量肽积累，每 20 克叶组织鲜重达到约 2.9 毫克 AMP。此外，纯化的 AMP 在人类原代皮肤细胞中表现出较低的附带毒性，通过透化膜杀死病原菌，并在临床前小鼠 (Mus musculus) 模型系统中表现出抗感染功效，将细菌负荷减少多达三个数量级。基本案例技术经济分析证明了我们基于工厂的平台的经济优势和可扩展性。我们设想我们的工作可以将植物建立为高效的生物反应器，用于以商业规模生产临床前级 AMP，并具有临床应用的潜力。