Evalution of tree frog antimicrobial peptide production by bacterial and plant expression system

Antimicrobial peptides play a role in innate immune system in all living organisms. Rhacoporin-2, a novel AMP from the skin secretion of Rhacophorus feae, shows an antimicrobial activity against both Gram-positive and Gram-negative bacteria. Contrastingly, it is relatively unharmful to mammalian red blood cell. Thus, it has a potential to be used as pharmaceutical agent. SUMO fusion system is employed for Rhacoporin-2 production. Therefore, the objective of this study is to evaluate the recombinant peptide, SUMO-Rhacoporin2, production by bacterial expression system and plant expression system. In Escherichia coli system, the total 131 µg/L of Rhacoporin2 with 85% purity was produced and the peptide exhibited the antibacterial activity against Salmonella Typhimurium at 8 µM. Hence, SUMO fusion system was effective for antimicrobial peptide production by bacterial system. For the recombinant peptide production in 2 plant hosts, Lemna minor and Arabidopsis thaliana, the results showed that the very low amount of recombinant peptide, 44.8 µg, was synthesized from 100 mg fresh duckweed, whereas, SUMOstar fusion tag for peptide production was unsuccessful in Arabidopsis. Therefore, SUMO fusion system was ineffective in these two species. However, plant system is still promising due to the low cost of production for therapeutic agents.抗菌肽（Antimicrobial peptides）在所有生物的先天免疫系统中发挥重要功能。蛙皮素-2（Rhacoporin-2）是一种源自费氏树蛙（Rhacophorus feae）皮肤分泌物的新型抗菌肽，对革兰氏阳性菌与革兰氏阴性菌均具有抗菌活性。与之相对的是，其对哺乳动物红细胞的细胞毒性相对较低，因此具备开发为药用制剂的潜力。本研究采用SUMO融合系统（SUMO fusion system）制备Rhacoporin-2，故本研究的核心目标是评估细菌表达系统与植物表达系统对重组肽SUMO-Rhacoporin2的制备效果。在大肠杆菌（Escherichia coli）表达系统中，最终获得了纯度为85%的Rhacoporin2，总产量达131 µg/L，且该肽在8 µM浓度下即可对鼠伤寒沙门氏菌（Salmonella Typhimurium）展现出抗菌活性。由此可见，SUMO融合系统可有效通过细菌表达系统制备抗菌肽。在两种植物宿主——小浮萍（Lemna minor）与拟南芥（Arabidopsis thaliana）中开展重组肽制备的实验结果显示：从100 mg新鲜小浮萍组织中仅合成了44.8 µg的重组肽；而在拟南芥中采用SUMOstar融合标签制备肽类的尝试并未取得成功。因此SUMO融合系统在这两种植物宿主中均未展现出应用有效性。不过鉴于治疗性制剂的生产成本较低，植物表达系统仍具备可观的应用前景。

搜集汇总

背景与挑战

背景概述

该数据集评估了树蛙抗菌肽（Rhacoporin-2）在细菌和植物表达系统中的生产效果。研究使用SUMO融合系统，发现在大肠杆菌中能高效生产具有抗菌活性的肽，而在两种植物宿主中产量较低，但植物系统因成本低仍具治疗剂生产潜力。数据集聚焦于抗菌肽的生物技术生产方法比较，涉及肽抗生素、Rhacophoridae等关键词。