Although most of antimicrobial peptides (AMPs), being relatively short, are produced by chemical synthesis, several AMPs have been produced using recombinant technology. However, AMPs could be cytotoxic to the producer cell, and if small they can be easily degraded. The objective of this study was to produce a multidomain antimicrobial protein based on recombinant protein nanoclusters to increase the yield, stability and effectivity. A single antimicrobial polypeptide JAMF1 that combines three functional domains based on human α-defensin-5, human XII-A secreted phospholipase A2 (sPLA2), and a gelsolin-based bacterial-binding domain along with two aggregation-seeding domains based on leucine zippers was successfully produced with no toxic effects for the producer cell and mainly in a nanocluster structure. Both, the nanocluster and solubilized format of the protein showed a clear antimicrobial effect against a broad spectrum of Gram-negative and Gram-positive bacteria, including multi-resistant strains, with an optimal concentration between 1 and 10 µM. Our findings demonstrated that multidomain antimicrobial proteins forming nanoclusters can be efficiently produced in recombinant bacteria, being a novel and valuable strategy to create a versatile, highly stable and easily editable multidomain constructs with a broad-spectrum antimicrobial activity in both soluble and nanostructured format.

中文翻译：

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新一代重组多肽结合了多个蛋白结构域，具有有效的抗菌活性。

尽管大多数较短的抗菌肽（AMPs）是通过化学合成产生的，但已经使用重组技术生产了几种AMPs。但是，AMPs对生产者细胞可能具有细胞毒性，如果很小，它们很容易降解。这项研究的目的是生产基于重组蛋白质纳米簇的多域抗菌蛋白，以提高产量，稳定性和有效性。一个单一的抗菌多肽JAMF1，结合了基于人α-防御素5的三个功能域，人XII-A分泌的磷脂酶A2（sPLA2）和基于凝溶胶蛋白的细菌结合域，以及两个基于亮氨酸拉链的聚集播种域已成功生产出对生产者细胞无毒且主要呈纳米簇结构的药物。都，蛋白质的纳米簇和可溶形式对多种革兰氏阴性和革兰氏阳性细菌（包括多重耐药菌株）表现出明显的抗菌作用，最佳浓度为1至10 µM。我们的研究结果表明，形成纳米簇的多域抗菌蛋白可以在重组细菌中高效生产，这是一种新颖且有价值的策略，可创建具有可溶和纳米结构形式的广谱抗菌活性的通用，高度稳定且易于编辑的多域构建体。