A rapid mutant screening technique for detection of technetium [Tc(VII)] reduction-deficient mutants ofShewanella oneidensisMR-1

Abstract

Microbial metal reduction forms the basis of alternate bioremediation strategies for reductive precipitation and immobilization of toxic metals such as the radionuclide technetium [Tc(VII)]. A rapid mutant screening technique was developed to identify Shewanella oneidensis MR-1 respiratory mutants unable to reduce Tc(VII) as anaerobic electron acceptor. The Tc(VII) reduction-deficient (Tcr) mutant screening technique was based on the observation that wild-type S. oneidensis produced a black Tc(IV) precipitate on its colony surface during growth on Tc(VII)-amended agar, while colonies arising from mutagenized cells did not. Tcr mutants unable to produce the black precipitate were subsequently tested for anaerobic growth on an array of three electron donors and 13 alternate electron acceptors. The Tcr mutants displayed a broad spectrum of anaerobic growth deficiencies, including several that were unable to reduce Tc(VII) with hydrogen or lactate as electron donor, yet retained the ability to reduce Tc(VII) with formate. This report describes the development of a novel Tcr mutant screening technique and its application to identify the first set of Tcr mutants in a metal-reducing member of the genus Shewanella.