Applied Biochemistry and Biotechnology

, Volume 175, Issue 3, pp 1732–1744 | Cite as

A Cysteine Protease Isolated from the Latex of Ficus microcarpa: Purification and Biochemical Characterization

  • Ibtissem Hamza Mnif
  • Rayda Siala
  • Rim Nasri
  • Samiha Mhamdi
  • Moncef Nasri
  • Alya Sellami Kamoun
Article
First Online:
Received:
Accepted:

Abstract

A plant protease named microcarpain was purified from the latex of Ficus microcarpa by acetonic (20–40 % saturation) precipitation, Sephadex G-75 filtration, and Mono Q-Sefinose FF chromatography. The protease was purified with a yield of 9.25 % and a purification factor of 8. The molecular weight of the microcarpain was estimated to be 20 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The purified enzyme showed maximum activity at pH 8.0 and at a temperature of 70 °C. Proteolytic activity was strongly inhibited by dithio-bis-nitrobenzoic acid (DTNB), Hg2+, and Cu2+. The N-terminal amino acid sequence of the purified microcarpain “VPETVDWRSKGAV” showed high homology with a protease from Arabidopsis thaliana. Inhibition studies and N-terminal sequence classified the enzyme as a member of the cysteine peptidases family.

Keywords

Ficus microcarpa latex Cysteine protease Purification Microcarpain 
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Notes

Acknowledgments

This work was funded by the Ministry of Higher Education and Scientific Research, Tunisia.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Ibtissem Hamza Mnif
    • 1
  • Rayda Siala
    • 1
  • Rim Nasri
    • 1
  • Samiha Mhamdi
    • 1
  • Moncef Nasri
    • 1
  • Alya Sellami Kamoun
    • 1
  1. 1.Laboratory of Enzyme Engineering and Microbiology, National School of Engineering of SfaxUniversity of SfaxSfaxTunisia

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