Extracytoplasmic storage as the nickel resistance mechanism in a natural isolate of Pseudomonas putida S4.

Metal resistances in microbes are important to study not only to understand metal homeostasis but also to use such organisms further in environmental bioremediation. Nickel (Ni2+) is an important micronutrient, which at higher concentration becomes toxic. Many Ni2+-resistant organisms are known, which resist metal by active efflux. Pseudomonas putida S4, a natural isolate from India, is reported to show a multi-metal resistance profile. In the present study, the Ni2+-resistance mechanism in strain S4 was examined. Wild-type cells gradually accumulated Ni2+ but kept it preferentially in the periplasmic space in a bound form. In Ni2+-sensitive mutants, periplasmic storage was disturbed and more metal accumulated cytoplasmically, producing toxicity. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis of periplasmic proteins revealed a band of approximately 18 kDa, which appeared only in Ni2+-exposed wild-type cells, and which was absent from cells not exposed to Ni2+ as well as from Ni2+-sensitive mutants. On the basis of these observations, we propose a Ni2+-resistance mechanism in P. putida S4 based on sequestration of metal in the periplasmic space. This is the first study of sequestration-based Ni2+ resistance.