Laboratory of Bacterial Genetics and Environmental Biotechnology, Department of Microbiology, Goa University, Taleigao Plateau, Goa, 403206, India.
Center of Advanced Study in Botany, Banaras Hindu University, Varanasi, U.P., 221005, India.
Biometals. 2019 Feb;32(1):65-76. doi: 10.1007/s10534-018-0158-7. Epub 2018 Nov 23.
Arsenite oxidizing Klebsiella pneumoniae strain SSSW7 isolated from shipyard waste Goa, India showed a minimum inhibitory concentration of 21 mM in mineral salts medium. The strain possessed a small supercoiled plasmid and PCR amplification of arsenite oxidase gene (aioA) was observed on plasmid as well as chromosomal DNA. It was confirmed that arsenite oxidase enzyme was a periplasmic protein with a 47% increase in arsenite oxidase activity at 1 mM sodium arsenite. Scanning electron microscopy coupled with electron dispersive X-ray spectroscopic (SEM-EDS) analysis of 15 mM arsenite exposed cells revealed long chains of cells with no surface adsorption of arsenic. Transmission electron microscopy combined with electron dispersive X-ray spectroscopic (TEM-EDS) analysis demonstrated plasma membrane disruption, cytoplasmic condensation and periplasmic accumulation of arsenic. The bacterial strain oxidized 10 mM of highly toxic arsenite to less toxic arsenate after 24 h of incubation. Fourier transformed infrared (FTIR) spectroscopy confirmed the interaction of arsenite with functional groups present on the bacterial cell surface. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of 5 mM arsenite exposed cells demonstrated over-expression of 87 kDa and 14 kDa proteins of two subunits aioA and aioB of heterodimer arsenite oxidase enzyme as compared to control cells. Therefore, this bacterial strain might be employed as a potential candidate for bioremediation of arsenite contaminated environmental sites.
从印度果阿船厂废物中分离出的亚砷酸盐氧化型肺炎克雷伯氏菌 SSSW7 在无机盐培养基中的最小抑菌浓度为 21mM。该菌株具有一个小的超螺旋质粒,并且在质粒和染色体 DNA 上都观察到亚砷酸盐氧化酶基因 (aioA) 的 PCR 扩增。证实亚砷酸盐氧化酶是一种周质蛋白,在 1mM 亚砷酸钠存在下,亚砷酸盐氧化酶活性增加了 47%。扫描电子显微镜结合电子能谱分析(SEM-EDS)对 15mM 亚砷酸盐暴露细胞的分析表明,细胞呈长链状,没有表面吸附砷。透射电子显微镜结合电子能谱分析(TEM-EDS)表明,细胞膜破裂,细胞质浓缩,周质积累砷。该菌株在孵育 24 小时后将 10mM 的高毒性亚砷酸盐氧化为毒性较低的砷酸盐。傅里叶变换红外(FTIR)光谱证实了亚砷酸盐与细菌细胞表面存在的官能团相互作用。与对照细胞相比,5mM 亚砷酸盐暴露细胞的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析表明,杂二聚体亚砷酸盐氧化酶的两个亚基 aioA 和 aioB 的 87kDa 和 14kDa 蛋白表达过度。因此,该菌株可能被用作生物修复受亚砷酸盐污染的环境地点的潜在候选菌株。