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基于 AlpB 外膜蛋白和胶体金/纳米多孔金框架的快速抗幽门螺杆菌生物膜药物筛选生物传感器。

A rapid anti-Helicobacter pylori biofilm drug screening biosensor based on AlpB outer membrane protein and colloidal gold/nanoporous gold framework.

机构信息

State Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, PR China.

Key Laboratory for Experimental Teratology of Chinese Ministry of Education, The Shandong Provincial Key Laboratory of Infection and Immunology, Department of Microbiology, School of Basic Medical Sciences, Shandong University, Jinan, 250012, PR China.

出版信息

Biosens Bioelectron. 2022 Nov 1;215:114599. doi: 10.1016/j.bios.2022.114599. Epub 2022 Jul 31.

Abstract

Inhibition or disruption of biofilms has been recognized as an important means to eradicate Helicobacter pylori (H. pylori) infection. However, a fast and efficient drug screening method against H. pylori biofilms has not yet been established. Therefore, AlpB, an important outer membrane protein in H. pylori biofilm formation, was selected as a biological recognition element to screen anti-biofilm drugs in this study. A novel AlpB/colloidal gold (CG)/nanoporous gold (NPG)/Nafion-reduced graphene oxide (rGO)/glassy carbon electrode (GCE) biosensor was constructed based on the heterologous expression of AlpB. The prepared AlpB-based biosensor not only successfully identified six anti-biofilm drugs, but also evaluated the sensitivity and action intensity of different anti-biofilm drugs binding to AlpB by interaction kinetics analysis. The sensitivity order of AlpB to the six anti-biofilm drugs was: allicin > erythromycin > SCC > curcumin > rifampicin > NAC and the action intensity of the six anti-biofilm drugs on AlpB was: rifampicin > NAC > allicin > erythromycin > SCC > curcumin. In addition, molecular docking results showed that the six anti-biofilm drugs might exert their anti-biofilm effects by spontaneously binding to the conserved region of AlpB protein. This study provided a rapid screening platform and a unified data processing method for potential anti-biofilm drug development.

摘要

抑制或破坏生物膜已被认为是根除幽门螺杆菌(H. pylori)感染的重要手段。然而,尚未建立针对 H. pylori 生物膜的快速有效的药物筛选方法。因此,本研究选择 H. pylori 生物膜形成过程中的重要外膜蛋白 AlpB 作为生物识别元件,筛选抗生物膜药物。基于 AlpB 的异源表达,构建了一种新型的 AlpB/胶体金(CG)/纳米多孔金(NPG)/Nafion-还原氧化石墨烯(rGO)/玻碳电极(GCE)生物传感器。所制备的基于 AlpB 的生物传感器不仅成功鉴定了六种抗生物膜药物,还通过相互作用动力学分析评估了不同抗生物膜药物与 AlpB 结合的敏感性和作用强度。AlpB 对六种抗生物膜药物的敏感性顺序为:大蒜素>红霉素>SCC>姜黄素>利福平>NAC,六种抗生物膜药物对 AlpB 的作用强度为:利福平>NAC>大蒜素>红霉素>SCC>姜黄素。此外,分子对接结果表明,六种抗生物膜药物可能通过自发结合 AlpB 蛋白的保守区域发挥其抗生物膜作用。本研究为潜在抗生物膜药物的开发提供了快速筛选平台和统一的数据处理方法。

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