设计,选和特征化工程化菌体内素与细胞外抗菌活性对抗阴性细菌的设计,选和特征化
Bingrui Sui1,2, Xinrui Wang1, Tianyi Zhao1
1College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, China.
Applied and environmental microbiology
|June 20, 2023
概括
改造的内素克服了格拉姆阴性细菌.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 蛋白质工程是指蛋白质工程.
背景情况:
- 菌体内素是一种强大的抗菌剂,可对抗格拉姆阳性细菌.
- 格拉姆阴性细菌的外膜阻碍了内素的透和活动.
- 工程内素对于提高其对抗格拉姆阴性病原体的有效性至关重要.
研究的目的:
- 开发和验证一个查平台的工程内素与细胞外抗菌活性对抗阴性细菌.
- 识别具有改进的透和溶解能力的新型嵌合体内素 (人工-Bp7e).
- 描述有希望的工程内素的特性和抗菌谱.
主要方法:
- 通过在Bp7e内素基因上游插入随机的NNK编码子来构建一个模拟内素库.
- 在大肠杆菌中表达化学蛋白质,并通过化雾化释放.
- 使用斑点和殖民地计数方法选活跃的内素,然后进行序列分析和选定候选者的表征.
主要成果:
- 成功选工程人工-Bp7e (Art-Bp7e) 具有细胞外活性的内分泌素,以对抗阴性细菌.
- 选的Art-Bp7e蛋白质具有具有正电荷和α-螺旋结构的化学.
- 一种代表性变体Art-Bp7e6显示出对大肠杆菌,沙门氏菌, Pseudomonas 和 Staphylococcus aureus 的广泛活性.
- 艺术-Bp7e6促进了格拉姆阴性细胞外去极化和提高透性,使得糖溶解.
结论:
- 开发的查平台有效地识别出具有细胞外活性的化学内素对抗格拉姆阴性细菌.
- 像Art-Bp7e6这样的工程内素可以克服Gram负细菌包裹所带来的挑战.
- 该平台提供了一种多功能工具,用于选工程蛋白质,在抗菌疗法及其他领域具有广泛的应用.
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