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Trypsin Inhibitor from Edible Mushroom Pleurotus floridanus Active against Proteases of Microbial Origin

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Abstract

Protease inhibitors can be versatile tools mainly in the fields of medicine, agriculture and food preservative applications. Fungi have been recognized as sources of protease inhibitors, although there are only few such reports on mushrooms. This work reports the purification and characterization of a trypsin inhibitor from the fruiting body of edible mushroom Pleurotus floridanus (PfTI) and its effect on the activity of microbial proteases. The protease inhibitor was purified up to 35-fold by DEAE-Sepharose ion exchange column, trypsin-Sepharose column and Sephadex G100 column. The isoelectric point of the inhibitor was 4.4, and its molecular mass was calculated as 37 kDa by SDS-PAGE and 38.3 kDa by MALDI-TOF. Inhibitory activity confirmation was by dot-blot analysis and zymographic activity staining. The specificity of the inhibitor toward trypsin was with Ki of 1.043 × 10−10 M. The inhibitor was thermostable up to 90 °C with maximal stability at 30 °C, active over a pH range of 4–10 against proteases from Aspergillus oryzae, Bacillus licheniformis, Bacillus sp. and Bacillus amyloliquefaciens. Results indicate the possibility of utilization of protease inhibitor from P. floridanus against serine proteases.

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Acknowledgments

One of the authors Manzur Ali P P is grateful to University Grants Commission for providing Teacher Fellowship. Financial support from Kerala Biotech Commission, KSCSTE, Kerala (Project Fellowship 739/MS/2011–2012 dated 19.03.2012) is gratefully acknowledged by the last author.

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Authors and Affiliations

  1. Department of Biotechnology, Cochin University of Science and Technology, Cochin, 682022, Kerala, India

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

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

    P. P. Manzur Ali

  3. Department of Botany and Microbiology, College of Science, King Saud University, PB No. 2455, Riyadh, 11451, Saudi Arabia

    M. Chandrasekaran

  4. Department of Biotechnology, Calicut University, Malappuram, 673635, Kerala, India

    K. K Elyas

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  1. P. P. Manzur Ali
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  2. K. Sapna
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  4. Sarita G Bhat
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  6. K. K Elyas
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Correspondence to P. P. Manzur Ali.

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Ali, P.P.M., Sapna, K., Mol, K.R.R. et al. Trypsin Inhibitor from Edible Mushroom Pleurotus floridanus Active against Proteases of Microbial Origin. Appl Biochem Biotechnol 173, 167–178 (2014). https://doi.org/10.1007/s12010-014-0826-1

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  • DOI: https://doi.org/10.1007/s12010-014-0826-1

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