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Smart Cleaning Properties of a Multi Tolerance Keratinolytic Protease from an Extremophilic Bacillus tequilensis hsTKB2: Prediction of Enzyme Modification Site

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Abstract

Bacillus tequilensis hsTKB2, produced extracellular alkaline (9–10.5), thermostable (50–80 °C), halophilic (0–30 %), organic solvent tolerant keratinolytic protease. The maximum enzyme production was observed in high alkaline condition (pH 10) at 45 °C in sea water as supporting medium and chicken feathers as substrate with cell-immobilization of chitin flakes. The molecular mass of purified enzyme was estimated to be about 59.89 kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF analysis. The apparent Km for the enzyme activity on keratin was 1.9 mg mL−1. Optimum temperature and pH for keratinolytic protease activity were 70 °C and 10.5 respectively. The effect of organic solvents on the enzyme activity showed that this enzyme was more stable in the presence of non-polar organic solvents than polar solvents. Circular dichroism spectroscopy revealed that at pH 10.5 and temperature 70 °C, the enzyme upholds a typical secondary structure (α-helix 25.23 %, β-sheet 61.02 %, turn 11.97 % and coil 1.78 %). Mn2+ and toluene were influenced the enzyme activity by effecting structural changes. This enzyme folded into its active conformation inside the cell and secreted outside. Furthermore the tolerability with surfactants and commercial solid and liquid detergents appeared to be a favorable approach towards laundry and dishwashing detergents.

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Acknowledgments

The authors gratefully acknowledge the financial support of UGC-RFSMS Fellowship scheme [F.11-114/2008 (BSR)] for present research.

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Authors and Affiliations

  1. Department of Microbiology, Vidyasagar University, Midnapore, 721102, West Bengal, India

    Tanmay Paul, Arpan Das, Arpita Mandal, Arijit Jana, Suman K. Halder, Pradeep K. Das Mohapatra, Bikas R. Pati & Keshab C. Mondal

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  1. Tanmay Paul
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  8. Keshab C. Mondal
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Correspondence to Keshab C. Mondal.

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Paul, T., Das, A., Mandal, A. et al. Smart Cleaning Properties of a Multi Tolerance Keratinolytic Protease from an Extremophilic Bacillus tequilensis hsTKB2: Prediction of Enzyme Modification Site. Waste Biomass Valor 5, 931–945 (2014). https://doi.org/10.1007/s12649-014-9310-y

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