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In vitro Cr(VI) reduction by cell-free extracts of chromate-reducing bacteria isolated from tannery effluent irrigated soil

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Abstract

Four efficient Cr(VI)-reducing bacterial strains were isolated from rhizospheric soil of plants irrigated with tannery effluent and investigated for in vitro Cr(VI) reduction. Based on 16S rRNA gene sequencing, the isolated strains SUCR44, SUCR140, SUCR186, and SUCR188 were identified as Bacillus sp. (JN674188), Microbacterium sp. (JN674183), Bacillus thuringiensis (JN674184), and Bacillus subtilis (JN674195), respectively. All four isolates could completely reduce Cr(VI) in culture media at 0.2 mM concentration within a period of 24–120 h; SUCR140 completely reduced Cr(VI) within 24 h. Assay with the permeabilized cells (treated with Triton X-100 and Tween 80) and cell-free assay demonstrated that the Cr(VI) reduction activity was mainly associated with the soluble fraction of cells. Considering the major amount of chromium being reduced within 24–48 h, these fractions could have been released extracellularly also during their growth. At the temperature optima of 28 °C and pH 7.0, the specific activity of Cr(VI) reduction was determined to be 0.32, 0.42, 0.34, and 0.28 μmol Cr(VI) min−1 mg−1 protein for isolates SUCR44, SUCR140, SUCR186, and SUCR188, respectively. Addition of 0.1 mM NADH enhanced the Cr(VI) reduction in the cell-free extracts of all four strains. The Cr(VI) reduction activity in cell-free extracts of all the isolates was stable in presence of different metal ions tested except Hg2+. Beside this, urea and thiourea also reduced the activity of chromate reduction to significant levels.

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Acknowledgments

The authors wish to thank the Director, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow, India, for providing necessary facilities and encouragement during the course of investigation, Dr HP Singh for his help in statistical analysis, Sukhmal Chand and Kundan Wasnik for soil analysis, and the Indian Council of Medical Research (ICMR), New Delhi, India, for providing financial support to SKS.

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Authors and Affiliations

  1. Department of Microbial Technology and Entomology, CSIR-Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), Lucknow, 226015, India

    Sumit K. Soni, Rakshapal Singh, Ashutosh Awasthi, Mangal Singh & Alok Kalra

  2. CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226015, Uttar Pradesh, India

    Alok Kalra

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  1. Sumit K. Soni
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  2. Rakshapal Singh
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Correspondence to Alok Kalra.

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Soni, S.K., Singh, R., Awasthi, A. et al. In vitro Cr(VI) reduction by cell-free extracts of chromate-reducing bacteria isolated from tannery effluent irrigated soil. Environ Sci Pollut Res 20, 1661–1674 (2013). https://doi.org/10.1007/s11356-012-1178-4

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