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Biophysicochemical Characterization of an Alkaline Protease from Beauveria sp. MTCC 5184 with Multiple Applications

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Abstract

This study illustrates the biophysicochemical properties of an alkaline protease, BAP (Beauveria sp. alkaline protease) from Beauveria sp. MTCC 5184. This protease exhibited maximum activity at 50 °C, pH 9.0, and stability in a broad pH range, in the presence of organic solvents, denaturants, as well as detergents. Wash performance studies revealed that BAP was able to remove blood clots/stains from blood-soaked cloth. Peptide mass fingerprinting results demonstrated partial homology of BAP with subtilisin-like proteinase. BAP showed catalytic activity against natural as well as synthetic substrates. Active site characterization of BAP confirmed the involvement of serine, tryptophan, and aspartic acid in catalytic activity. Detailed kinetic and thermodynamic studies of BAP demonstrated that the activation energy (Ea) for casein hydrolysis was 82.55 kJ/M, the specificity constant (Kcat/K m), and the values of ∆G (change in Gibbs free energy) decreased with increase in temperature, whereas ∆H (change in enthalapy) and ∆S (change in entropy) were constant. The results of the present study indicate that BAP has potential for applications as detergent additive, in peptide synthesis, and in basic research.

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Acknowledgments

Shiv Shankar is grateful to the Council of Scientific and Industrial Research (CSIR) for financial assistance. Authors acknowledge the financial support from NMITLI project funded by CSIR, Govt. of India, and Dr. Mahesh J Kulkarni for analysis of samples in MALDI.

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Authors and Affiliations

  1. Division of Biochemical Sciences, National Chemical Laboratory, Pune, 411 008, India

    Shiv Shankar & Ryali Seeta Laxman

  2. Department of Food Engineering, Mokpo National University, 61 Dorimri, Chungkyemyon, Muangun, Jeonnam, 534729, Republic of Korea

    Shiv Shankar

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  1. Shiv Shankar
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  2. Ryali Seeta Laxman
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Correspondence to Shiv Shankar or Ryali Seeta Laxman.

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Shankar, S., Laxman, R.S. Biophysicochemical Characterization of an Alkaline Protease from Beauveria sp. MTCC 5184 with Multiple Applications. Appl Biochem Biotechnol 175, 589–602 (2015). https://doi.org/10.1007/s12010-014-1314-3

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