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Biological characterization of Bacillus flexus strain SSAI1 transforming highly toxic arsenite to less toxic arsenate mediated by periplasmic arsenite oxidase enzyme encoded by aioAB genes

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Abstract

Bacillus flexus strain SSAI1 isolated from agro-industry waste, Tuem, Goa, India displayed high arsenite resistance as minimal inhibitory concentration was 25 mM in mineral salts medium. This bacterial strain exposed to 10 mM arsenite demonstrated rapid arsenite oxidation and internalization of 7 mM arsenate within 24 h. The Fourier transformed infrared (FTIR) spectroscopy of cells exposed to arsenite revealed important functional groups on the cell surface interacting with arsenite. Furthermore, scanning electron microscopy combined with electron dispersive X-ray spectroscopy (SEM-EDAX) of cells exposed to arsenite revealed clumping of cells with no surface adsorption of arsenite. Transmission electron microscopy coupled with electron dispersive X-ray spectroscopic (TEM-EDAX) analysis of arsenite exposed cells clearly demonstrated ultra-structural changes and intracellular accumulation of arsenic. Whole-genome sequence analysis of this bacterial strain interestingly revealed the presence of large number of metal(loid) resistance genes, including aioAB genes encoding arsenite oxidase responsible for the oxidation of highly toxic arsenite to less toxic arsenate. Enzyme assay further confirmed that arsenite oxidase is a periplasmic enzyme. The genome of strain SSAI1 also carried glpF, aioS and aioE genes conferring resistance to arsenite. Therefore, multi-metal(loid) resistant arsenite oxidizing Bacillus flexus strain SSAI1 has potential to bioremediate arsenite contaminated environmental sites and is the first report of its kind.

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Acknowledgements

Ms. Sajiya Yusuf Mujawar is grateful to University Grants Commission, New Delhi for financial support as MANF-SRF (Award Letter No. F1-17.1/2015-16/MANF-2015-17-Goa-50641). The authors are thankful to Mr. Areef Sardar from CSIR-National Institute of Oceanography, Goa for EDAX analysis and AIRF, Jawaharlal Nehru University, New Delhi for TEM-EDAX analysis. The authors are also obliged to Dr. B. R. Srinivasan and Mr. Rahul Kerkar from Department of Chemistry, Goa University for FTIR analysis. The authors are also grateful to Dr. Sandeep Garg, Head, Department of Microbiology, Goa University for providing laboratory facilities.

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  1. Laboratory of Bacterial Genetics and Environmental Biotechnology, Department of Microbiology, Goa University, Taleigao Plateau, Goa, 403206, India

    Sajiya Yusuf Mujawar, Diviya Chandrakant Vaigankar & Santosh Kumar Dubey

  2. Center of Advanced Study in Botany, Banaras Hindu University, Varanasi, Uttar Pradesh, 221005, India

    Santosh Kumar Dubey

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  1. Sajiya Yusuf Mujawar
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SYM: Conceptualization, experiments, data generation, analysis and writing of the original draft manuscript. DCV: Assisted in experimental work and draft writing. SKD: Experimental designs, verification of data, mentoring of experiments, correction and final editing of the research manuscript.

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Correspondence to Santosh Kumar Dubey.

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Mujawar, S.Y., Vaigankar, D.C. & Dubey, S.K. Biological characterization of Bacillus flexus strain SSAI1 transforming highly toxic arsenite to less toxic arsenate mediated by periplasmic arsenite oxidase enzyme encoded by aioAB genes. Biometals 34, 895–907 (2021). https://doi.org/10.1007/s10534-021-00316-x

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