Department of Biotechnology, University of Pune, Ganeshkhind, Pune, 411007, India.
Appl Microbiol Biotechnol. 2014 Aug;98(15):6761-73. doi: 10.1007/s00253-014-5873-6. Epub 2014 Jun 17.
The general responses of microorganisms to environmental onslaughts are modulated by altering the gene expression pattern to reduce damage in the cell and produce compensating stress responses. The present study attempts to unravel the response of the Gram-positive Exiguobacterium sp. PS NCIM 5463 in the presence of [As(III)] and arsenate [As(V)] using comparative proteomics via two-dimension gel electrophoresis (2-DE) coupled with identification of proteins using matrix-assisted laser desorption/ionisation (MALDI-TOF/MALDI-TOF/TOF). Out of 926 Coomassie-stained proteins, 45 were differentially expressed (p < 0.05). Considering the resolution and abundance level, 24 spots (peptides) were subjected to MALDI analysis, identified and categorised into several functional categories, viz., nitrogen metabolism, energy and stress regulators, carbohydrate metabolism, protein synthesis components and others. A functional role of each protein is discussed in Exiguobacterium sp. PS 5463 under arsenic stress and validated at their transcript level using a quantitative real-time polymerase chain reaction. Unlike previous reports that unravel the responses toward arsenic stress in Gram-negative organisms, the present study identified new proteins under arsenic stress in a Gram-positive organism, Exiguobacterium sp. PS NCIM 5463, which could elucidate the physiology of organisms under arsenic stress.
微生物对环境侵袭的一般反应是通过改变基因表达模式来调节的,以减少细胞损伤并产生补偿应激反应。本研究试图通过二维凝胶电泳 (2-DE) 结合基质辅助激光解吸/电离 (MALDI-TOF/MALDI-TOF/TOF) 鉴定蛋白质,来揭示革兰氏阳性极端微生物 Exiguobacterium sp. PS NCIM 5463 在 [As(III)] 和砷酸盐 [As(V)] 存在下的反应。在 926 个考马斯亮蓝染色的蛋白质中,有 45 个表现出差异表达(p<0.05)。考虑到分辨率和丰度水平,对 24 个斑点(肽)进行 MALDI 分析,并将其分类为几个功能类别,即氮代谢、能量和应激调节剂、碳水化合物代谢、蛋白质合成成分等。在砷胁迫下,讨论了每个蛋白质在 Exiguobacterium sp. PS 5463 中的功能作用,并通过定量实时聚合酶链反应在转录水平上进行了验证。与以前的研究不同,这些研究揭示了革兰氏阴性生物对砷胁迫的反应,本研究在革兰氏阳性生物 Exiguobacterium sp. PS NCIM 5463 中鉴定出了砷胁迫下的新蛋白质,这可以阐明砷胁迫下生物体的生理学。