Prokaryotic 表达和功能验证的抗微生物LRGG的功能验证
Xiang Liu1, Yining Ding1, Yuhan Shen1
1Engineering Research Center, The Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, College of Life Sciences, Jilin Agricultural University, Changchun 130118, China.
International journal of molecular sciences
|July 13, 2024
概括
Prokaryotic 表达产生一种抗微生物,LRGG,有效对抗阴性细菌. 这种方法提供了一个稳定的,无毒的治疗方法,具有广泛的抗菌活性.
科学领域:
- 生物技术是生物技术.
- 微生物学 微生物学
- 酸科学 酸科学
背景情况:
- 抗微生物 (AMP) 在天生的免疫力中至关重要.
- LRGG是一种合成AMP,显示出对抗格拉姆阴性细菌的前景.
- 为AMP开发具有成本效益和可扩展的生产方法至关重要.
研究的目的:
- 为抗微生物LRGG.建立一个 prokaryotic表达系统.
- 为了比较 prokaryotic 表达的 LRGG 与化学合成的 LRGG 的特性和有效性.
- 研究LRGG的作用机制和治疗潜力.
主要方法:
- LRGG的设计和化学合成.
- 构建一个 prokaryotic 核融合表达向量 (pQE-GFP-LRGG).
- 融合蛋白GFP-LRGG的表达,净化和内毒素去除.
- 在TEV蛋白质溶解中释放纯LRGG.
- 对LRGG稳定性 (温度,pH,盐,血清) 和细胞选择性的表征.
- 在Galleria mellonella感染模型中评估治疗疗效.
主要成果:
- Prokaryotic 的表达成功地产生了纯 LRGG.
- LRGG证明了针对格拉姆阴性细菌的广泛活性,最低抑制度 (MIC) 为2μg/mL对抗大肠杆菌.
- Prokaryotic-expressed LRGG 呈现出与其化学合成对应物相比的可比稳定性和细胞选择性.
- 没有观察到胚胎毒性,并在Galleria mellonella感染模型中观察到出色的治疗效果.
- 作用的机制涉及破坏格拉姆阴性细菌细胞膜.
结论:
- Prokaryotic 表达是一种生产功能性抗微生物 LRGG.GG 的可行方法.
- Prokaryotic-expressed LRGG 是一种强效,稳定和安全的治疗候选物,可以对抗格拉姆阴性细菌感染.
- LRGG的膜破坏机制为打击抗生素耐药细菌提供了一个有希望的途径.
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