Abstract
Arsenic is a naturally occurring ubiquitous highly toxic metalloid. In this study, we have identified ars gene cluster in Pannonibacter indicus strain HT23T (DSM 23407T), responsible for reduction of toxic pentavalent arsenate. The ars gene cluster is comprised of four non-overlapping open reading frames (ORFs) encoding a transcriptional regulator (ArsR), a low molecular weight protein tyrosine phosphatases (LMW-PTPase) with hypothetical function, an arsenite efflux pump (Acr3), and an arsenate reductase (ArsC). Heterologous expression of arsenic inducible ars gene cluster conferred arsenic resistance to Escherichia coli ∆ars mutant strain AW3110. The recombinant ArsC was purified and assayed. Site-directed mutagenesis was employed to ascertain the role of specific amino acids in ArsC catalysis. Pro94X (X = Ala, Arg, Cys, and His) amino acid substitutions led to enzyme inactivation. Circular dichroism spectra analysis suggested Pro94 as an essential amino acid for enzyme catalytic activity as it is indispensable for optimum protein folding in P. indicus Grx-coupled ArsC.
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Acknowledgments
We are grateful to the Department of Chemistry, Indian Institute of Technology, Bhubaneswar, for the CD spectroscopic analysis. This work was supported in part by the funding received from Department of Biotechnology, Government of India (D.O. No. BT/PR9712/NBD/52/91/2007) to SKD. The author SB acknowledges the Council of Scientific and Industrial Research, Government of India, for providing the research fellowship. We like to thank Dr. BK Mohanty (University of Georgia, USA) for critically reading the manuscript.
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Bandyopadhyay, S., Das, S.K. Functional analysis of ars gene cluster of Pannonibacter indicus strain HT23T (DSM 23407T) and identification of a proline residue essential for arsenate reductase activity. Appl Microbiol Biotechnol 100, 3235–3244 (2016). https://doi.org/10.1007/s00253-016-7390-2
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DOI: https://doi.org/10.1007/s00253-016-7390-2