Bar Chinmayee, Patil Rajendra, Doshi Jignesh, Kulkarni Mahesh J, Gade W N
Department of Biotechnology, University of Pune, Pune 411007, India.
J Biotechnol. 2007 Feb 20;128(3):444-51. doi: 10.1016/j.jbiotec.2006.11.010. Epub 2006 Nov 24.
The present study describes response of a bacterial strain isolated from a polluted river to heavy metal toxicity. The bacterium was identified to be Klebsiella pneumoniae by biochemical tests using API 20E strips and 16S ribotyping. The isolate was studied for its tolerance to two heavy metals, i.e., cobalt (Co(2+)) and lead (Pb(2+)) by growing it in citrate mineral medium (CMM). Proteomic approach involving two-dimensional polyacrylamide gel electrophoresis (2D PAGE) and mass spectrometry (MS) was used to identify the differentially expressed proteins under heavy metal stress. Two of the differentially expressed proteins were identified to be l-isoaspartate protein carboxymethyltransferase type II and DNA gyrase A. To our knowledge, this is for the first time that K. pneumoniae has been reported to be present in metal contaminated site and l-isoaspartate protein carboxymethyltransferase type II protein to be over expressed under heavy metal stress. The role of these proteins in metal tolerance is discussed.
本研究描述了从一条受污染河流中分离出的一种细菌菌株对重金属毒性的反应。通过使用API 20E试纸条进行生化试验和16S核糖体分型,鉴定该细菌为肺炎克雷伯菌。通过在柠檬酸盐矿物培养基(CMM)中培养该分离株,研究了其对两种重金属,即钴(Co(2+))和铅(Pb(2+))的耐受性。采用二维聚丙烯酰胺凝胶电泳(2D PAGE)和质谱(MS)的蛋白质组学方法来鉴定在重金属胁迫下差异表达的蛋白质。鉴定出两种差异表达的蛋白质为II型L-异天冬氨酸蛋白羧甲基转移酶和DNA促旋酶A。据我们所知,这是首次报道肺炎克雷伯菌存在于金属污染位点,以及II型L-异天冬氨酸蛋白羧甲基转移酶在重金属胁迫下过度表达。讨论了这些蛋白质在金属耐受性中的作用。