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Cytosolic expression of synthetic phytochelatin and bacterial metallothionein genes in Deinococcus radiodurans R1 for enhanced tolerance and bioaccumulation of cadmium

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Abstract

Due to its exemplary resistance to ionising radiation, oxidative stress, desiccation and several DNA damaging agents, Deinococcus radiodurans R1 (DR1) is considered as one of the most appropriate candidates for the bioremediation of the nuclear waste sites. However, the high sensitivity of this bacterium to heavy metals, which are usually preponderant at nuclear waste dump sites, precludes its application for bioremediation. This study deals with the expression two metal binding peptides in DR1 as an attractive strategy for developing metal tolerance in this bacterium. A synthetic gene (EC20) encoding a phytochelatin analogue with twenty repeating units of glutamate and cysteine was constructed by overlap extension and expressed in DR1. The cyanobacterial metallothionein (MT) gene, smtA was cloned for intracellular expression in DR1. Both the genes were expressed under the native groESL promoter. DR1 strain carrying the recombinant EC20 demonstrated 2.5-fold higher tolerance to Cd2+ and accumulated 1.21-fold greater Cd2+ as opposed to the control while the heterologous expression of MT SmtA in DR1 imparted the transformant superior tolerance to Cd2+ amassing 2.5-fold greater Cd2+ than DR1 expressing EC20.

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Acknowledgments

Fellowship to R. C. from Council of Scientific and Industrial Research, Government of India is gratefully acknowledged. R. C. is thankful to Dr. Priya Pillai for her assistance.

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Authors and Affiliations

  1. Department of Microbiology and Biotechnology Centre, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara, 390 002, Gujarat, India

    Ruchi Chaturvedi & G. Archana

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  1. Ruchi Chaturvedi
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  2. G. Archana
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Correspondence to G. Archana.

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Chaturvedi, R., Archana, G. Cytosolic expression of synthetic phytochelatin and bacterial metallothionein genes in Deinococcus radiodurans R1 for enhanced tolerance and bioaccumulation of cadmium. Biometals 27, 471–482 (2014). https://doi.org/10.1007/s10534-014-9721-z

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  • DOI: https://doi.org/10.1007/s10534-014-9721-z

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