Purification and characterization of a cathepsin L-like enzyme from the body wall of the sea cucumber Stichopus japonicus

Bei Wei Zhu, Lu Lu Zhao, Li Ming Sun, Dong Mei Li, Yoshiyuki Murata, Lei Yu, Lei Zhang

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Cathepsin L-like enzyme was purified from the body wall of the sea cucumber Stichopus japonicus by an integral method involving ammonium sulfate precipitation and a series of column chromatographies on DEAE Sepharose CL-6B, Sephadex G-75, and TSK-GEL. The molecular mass of the purified enzyme was estimated to be 63 kDa by SDS-PAGE. The enzyme cleaved N-carbobenzoxy- phenylalanine-arginine 7-amido-4-methylcoumarin with Km (69.92 μM) and kcat (12.80/S) hardly hydrolyzed N-carbobenzoxy-arginine- arginine 7-amido-4-methylcoumarin and L-arginine 7-amido-4-methylcoumarin. The optimum pH and temperature for the purified enzyme were found to be 5.0 and 50°C. It showed thermal stability below 40°C. The activity was inhibited by sulfhydryl reagents and activated by reducing agents. These results suggest that the purified enzyme was a cathepsin L-like enzyme and that it existed in the form of its enzyme-inhibitor complex or precursor.

Original languageEnglish
Pages (from-to)1430-1437
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume72
Issue number6
DOIs
Publication statusPublished - 2008

Keywords

  • Cathepsin L
  • Characterization
  • Protease
  • Purification
  • Sea cucumber (Stichopus japonicus)

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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