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Optimization, purification and characterization of novel thermostable, haloalkaline, solvent stable protease from Bacillus halodurans CAS6 using marine shellfish wastes: a potential additive for detergent and antioxidant synthesis

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Abstract

A protease producing marine bacterium, Bacillus halodurans CAS6 isolated from marine sediments, was found to produce higher enzyme by utilizing shrimp shell powder. Optimum culture conditions for protease production were 50 °C, pH 9.0, 30 % NaCl and 1 % shrimp shell powder (SSP) and the protease purified with a specific activity of 509.84 U/mg. The enzyme retained 100 % of its original activity even at 70 °C, pH 10.0 and 30 % NaCl for 1 h. The purified protease exhibited higher stability when treated with ionic, non-ionic (72–94 %) and commercial detergents (76–88 %), and organic solvents (88–126 %). Significant blood stain removal activity was found with the enzyme in washing experiments. The culture supernatant supplemented with 1 % SSP showed 93.67 ± 2.52 % scavenging activity and FT-IR analysis of the reaction mixture confirmed the presence of antioxidants such as cyclohexane and cyclic depsipeptide with aliphatic amino groups. These remarkable qualities found with this enzyme produced by Bacillus halodurans CAS6 could make this as an ideal candidate to develop the industrial process for bioconversion of marine wastes and antioxidant synthesis.

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Acknowledgments

We thank the authorities of Annamalai University for providing facilities and Ministry of Earth Sciences—Coastal Ocean Monitoring and Prediction System (MoES-COMAPS), Government of India for providing financial support.

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Authors and Affiliations

  1. Department of Chemistry, University of Puerto Rico at Cayey, 205, Ave. Antonio R. Barcelo, Cayey, PR, 00736, USA

    Neelamegam Annamalai

  2. CAS in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai, 608502, TN, India

    Mayavan Veeramuthu Rajeswari, Shanmugam Vijayalakshmi & Thangavel Balasubramanian

  3. Department of Chemical and Biological Sciences, Polytechnic Institute of New York University, 6 Metrotech Center, Brooklyn, NY, 11201, USA

    Rengathavasi Thavasi

Authors
  1. Neelamegam Annamalai
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  2. Mayavan Veeramuthu Rajeswari
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  3. Rengathavasi Thavasi
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  4. Shanmugam Vijayalakshmi
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  5. Thangavel Balasubramanian
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Correspondence to Neelamegam Annamalai.

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Annamalai, N., Rajeswari, M.V., Thavasi, R. et al. Optimization, purification and characterization of novel thermostable, haloalkaline, solvent stable protease from Bacillus halodurans CAS6 using marine shellfish wastes: a potential additive for detergent and antioxidant synthesis. Bioprocess Biosyst Eng 36, 873–883 (2013). https://doi.org/10.1007/s00449-012-0820-3

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  • DOI: https://doi.org/10.1007/s00449-012-0820-3

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