Abstract
The present study characterized aresenate reductase of Bacillus thuringiensis KPWP1, tolerant to salt, arsenate and a wide range of pH during growth. Interestingly, it was found that arsC, arsB and arsR genes involved in arsenate tolerance are distributed in the genome of strain KPWP1. The inducible arsC gene was cloned, expressed and the purified ArsC protein showed profound enzyme activity with the KM and Kcat values as 25 µM and 0.00119 s−1, respectively. In silico studies revealed that in spite of 19–26% differences in gene sequences, the ArsC proteins of Bacillus thuringiensis, Bacillus subtilis and Bacillus cereus are structurally conserved and ArsC structure of strain KPWP1 is close to nature. Docking and analysis of the binding site showed that arsenate ion interacts with three cysteine residues of ArsC and predicts that the ArsC from B. thuringiensis KPWP1 reduces arsenate by using the triple Cys redox relay mechanism.
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Acknowledgements
The authors acknowledge the service of SGS laboratory, Kolkata for atomic absorption spectroscopy facility. We also acknowledge the service of 1st BASE for DNA sequencing. Authors acknowledge Arpita Karmakar for technical assistance. Authors also like to thank Dr. Sumita Sengupta (Calcutta University) for her valuable suggestions regarding the manuscript writing. PB was a recipient of a Senior Research Fellowship from the Council of Scientific and Industrial Research, Government of India. AC, NB and SJ were recipients of Research Fellowship from Indian Institute of Science Education and Research (IISER) Kolkata. The work was supported by IISER Kolkata.
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PB, AC, PPD and TKS conceived the study and designed the experiments. PB, AC, NB and SJ performed the experiments and analysed the data. PB and TKS have written the manuscript.
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Banerjee, P., Chatterjee, A., Jha, S. et al. Biochemical, molecular and in silico characterization of arsenate reductase from Bacillus thuringiensis KPWP1 tolerant to salt, arsenic and a wide range of pH. Arch Microbiol 204, 46 (2022). https://doi.org/10.1007/s00203-021-02660-5
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DOI: https://doi.org/10.1007/s00203-021-02660-5