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Isolation, purification and characterization of a pH tolerant and temperature stable proteinaceous protease inhibitor from marine Pseudomonas mendocina

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Abstract

Objectives

An extracellular protease inhibitor (BTPI-301) of trypsin was purified and characterized from an isolate of Pseudomonas mendocina.

Results

BTPI-301was purified to homogeneity by (NH4)2SO4, precipitation, DEAE Sepharose and CNBr-activated Sepharose chromatography. Homogeneity was proved by native PAGE and SDS-PAGE. The intact molecular mass was 11567 Da by MALDI-TOF analysis. BTPI-301was a competitive inhibitor with a Ki of 3.5 × 10−10 M. It was stable and active at pH 4–12 and also at 4–90 °C for 1 h. Peptide mass fingerprinting by MALDI revealed that the BTPI-301 is a new inhibitor not reported so far with protease inhibitory activity. The pI of the inhibitor was 3.8. The stoichiometry of trypsin-BTPI-301 interaction is 1:1. The inhibitor was specific towards trypsin.

Conclusion

A pH tolerant and thermostable protease inhibitor BTPI-301 active against trypsin was purified and characterized from P. mendocina that could be developed and used as biopreservative as well as biocontrol agent.

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Acknowledgements

The authors gratefully acknowledge the financial support from Department of Biotechnology, Government of India (Sanction Order No.: BT/PR7906/AAQ/03/281/2006 dated 07.03.2007).

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Authors and Affiliations

  1. Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, 682 022, India

    K. Sapna, K. R. Rekha Mol, Sarita G. Bhat & M. Chandrasekaran

  2. Department of Biotechnology, MES College, Marampally, Aluva, Kerala, 683107, India

    P. P. Manzur Ali

  3. Department of Biotechnology, Calicut University, Malappuram, 673 635, Kerala, India

    K. K. Elyas

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  1. K. Sapna
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  2. P. P. Manzur Ali
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  3. K. R. Rekha Mol
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  4. Sarita G. Bhat
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  5. M. Chandrasekaran
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  6. K. K. Elyas
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Correspondence to K. Sapna.

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Sapna, K., Manzur Ali, P.P., Rekha Mol, K.R. et al. Isolation, purification and characterization of a pH tolerant and temperature stable proteinaceous protease inhibitor from marine Pseudomonas mendocina . Biotechnol Lett 39, 1911–1916 (2017). https://doi.org/10.1007/s10529-017-2424-0

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  • DOI: https://doi.org/10.1007/s10529-017-2424-0

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