Decoding the role of hypothetical protein All3255 of Anabaena PCC7120 in heavy metal stress management in Escherichia co
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ORIGINAL PAPER
Decoding the role of hypothetical protein All3255 of Anabaena PCC7120 in heavy metal stress management in Escherichia coli Prashant Kumar Singh1,2 · Mengmeng Tang1 · Sudhir Kumar2 · Alok Kumar Shrivastava3 Received: 8 September 2017 / Revised: 20 November 2017 / Accepted: 23 November 2017 © Springer-Verlag GmbH Germany, part of Springer Nature 2017
Abstract Cadmium is a non-essential toxic heavy metal for organisms, including plants and cyanobacteria. Cadmium resistance transporters involved in resistance of cells against various toxicants such as drugs and effluxes cytotoxic compounds from cells. However, cadmium resistance-associated protein (CadD) has never been reported from a diazotrophic cyanobacterium Anabaena sp. To test whether the hypothetical protein All3255 of Anabaena sp. PCC7120 a homolog of cadmium resistance-associated protein (CadD) involved in cadmium or heavy metal resistance or not, cloning and heterologous expression analysis of all3255 performed in Escherichia coli BL21 (DE3). Our results revealed that the strain transformed with pGEX-5X-2 + all3255 showed resistant towards not only to cadmium but also other heavy metals such as nickel, copper, zinc, lead and cobalt in addition to arsenic than those of transformed with empty vector (pGEX-5X-2). Furthermore, the results of metal accumulation analysis of these cells unveil a lower accumulation of tested heavy metals in all3255-overexpressing E. coli cells than those transformed with empty vector. This study strongly supports the role of All3255 of Anabaena sp. PCC7120 as a CadD efflux pump of heavy metals in E.coli. Keywords Anabaena sp. PCC7120 · Cadmium resistance transporter (CadD) · Escherichia coli BL21 (DE3) · Heavy metals · Hypothetical proteins · Resistance Abbreviations CadD Cadmium resistant transporter IPTG Isopropyl β-d-1-thiogalactopyranoside ROS Reactive oxygen species pI Isoelectric point Mw Molecular weight PCR Polymerase chain reaction
Communicated by Erko Stackebrandt. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00203-017-1462-2) contains supplementary material, which is available to authorized users. * Alok Kumar Shrivastava [email protected] 1
State Key Laboratory of Cotton Biology, Henan Key Laboratory of Plant Stress Biology, Henan University, Kaifeng 475004, Henan, China
2
Department of Botany, Banaras Hindu University, Varanasi, Uttar Pradesh 221005, India
3
Department of Botany, Mahatma Gandhi Central University, Motihari, Bihar 845401, India
RT-PCR Reverse transcriptase PCR qRT-PCR Quantitative real-time polymerase chain reaction
Introduction Anthropogenic activities in combination with modern agricultural practices pushing not only developing countries but also the developed one in one hand into an alarming state of heavy metal pollution while on the other reducing productivity of agriculturally fertile land (Valdman et al. 2001). It is worthwhile to mention that our waterbodies, as well as groundwater resources, ar
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