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多重耐药鲍曼不动杆菌AIIMS 7中algC基因表达模式的表征及其与非生物表面生物膜形成的相关性

Characterization of the algC gene expression pattern in the multidrug resistant Acinetobacter baumannii AIIMS 7 and correlation with biofilm development on abiotic surface.

作者信息

Sahu Praveen K, Iyer Pavithra S, Barage Sagar H, Sonawane Kailas D, Chopade Balu A

机构信息

Institute of Bioinformatics and Biotechnology, University of Pune, Pune 411 007, India ; Ispat General Hospital, SAIL, Rourkela 769 005, India.

Institute of Bioinformatics and Biotechnology, University of Pune, Pune 411 007, India.

出版信息

ScientificWorldJournal. 2014;2014:593546. doi: 10.1155/2014/593546. Epub 2014 Dec 3.

Abstract

Relative quantification of algC gene expression was evaluated in the multidrug resistant strain Acinetobacter baumannii AIIMS 7 biofilm (3 to 96 h, on polystyrene surface) compared to the planktonic counterparts. Comparison revealed differential algC expression pattern with maximum 81.59-fold increase in biofilm cells versus 3.24-fold in planktonic cells (P < 0.05). Expression levels strongly correlated with specific biofilm stages (scale of 3 to 96 h), coinciding maximum at initial surface attachment stage (9 h) and biofilm maturation stage (48 h). Cloning, heterologous expression, and bioinformatics analyses indicated algC gene product as the bifunctional enzyme phosphomannomutase/phosphoglucomutase (PMM/PGM) of ∼ 53 kDa size, which augmented biofilms significantly in algC clones compared to controls (lacking algC gene), further localized by scanning electron microscopy. Moreover, molecular dynamics analysis on the three-dimensional structure of PMM/PGM (simulated up to 10 ns) revealed enzyme structure as stable and similar to that in P. aeruginosa (synthesis of alginate and lipopolysaccharide core) and involved in constitution of biofilm EPS (extracellular polymeric substances). Our observation on differential expression pattern of algC having strong correlation with important biofilm stages, scanning electron-microscopic evidence of biofilm augmentation taken together with predictive enzyme functions via molecular dynamic (MD) simulation, proposes a new basis of A. baumannii AIIMS 7 biofilm development on inanimate surfaces.

摘要

与浮游态对应物相比,评估了多药耐药菌株鲍曼不动杆菌AIIMS 7生物膜(在聚苯乙烯表面上培养3至96小时)中algC基因表达的相对定量。比较发现algC表达模式存在差异,生物膜细胞中最大增加81.59倍,而浮游细胞中为3.24倍(P < 0.05)。表达水平与特定生物膜阶段(3至96小时范围)密切相关,在初始表面附着阶段(9小时)和生物膜成熟阶段(48小时)达到最大值。克隆、异源表达和生物信息学分析表明,algC基因产物是一种大小约为53 kDa的双功能酶磷酸甘露糖变位酶/磷酸葡萄糖变位酶(PMM/PGM),与对照(缺乏algC基因)相比,algC克隆中的生物膜显著增加,通过扫描电子显微镜进一步定位。此外,对PMM/PGM三维结构的分子动力学分析(模拟长达10纳秒)显示,该酶结构稳定,与铜绿假单胞菌(藻酸盐和脂多糖核心的合成)中的结构相似,并参与生物膜胞外聚合物(EPS)的构成。我们关于algC差异表达模式与重要生物膜阶段密切相关的观察结果,结合生物膜增加的扫描电子显微镜证据以及通过分子动力学(MD)模拟预测的酶功能,为鲍曼不动杆菌AIIMS 7在无生命表面上生物膜的形成提出了新的基础。

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