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

31 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

Fingerprint

Stichopus
Sea Cucumbers
Apostichopus japonicus
cathepsin L
Cathepsin L
Holothuroidea
integument
Purification
Enzymes
Arginine
arginine
enzymes
Sulfhydryl Reagents
Column chromatography
Enzyme Precursors
reducing agents
enzyme inhibitors
Reducing Agents
Ammonium Sulfate
Molecular mass

Keywords

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

ASJC Scopus subject areas

  • Bioengineering
  • Biotechnology
  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry (miscellaneous)
  • Applied Microbiology and Biotechnology
  • Food Science

Cite this

Purification and characterization of a cathepsin L-like enzyme from the body wall of the sea cucumber Stichopus japonicus. / Zhu, Bei Wei; Zhao, Lu Lu; Sun, Li Ming; Li, Dong Mei; Murata, Yoshiyuki; Yu, Lei; Zhang, Lei.

In: Bioscience, Biotechnology and Biochemistry, Vol. 72, No. 6, 2008, p. 1430-1437.

Research output: Contribution to journalArticle

Zhu, Bei Wei ; Zhao, Lu Lu ; Sun, Li Ming ; Li, Dong Mei ; Murata, Yoshiyuki ; Yu, Lei ; Zhang, Lei. / Purification and characterization of a cathepsin L-like enzyme from the body wall of the sea cucumber Stichopus japonicus. In: Bioscience, Biotechnology and Biochemistry. 2008 ; Vol. 72, No. 6. pp. 1430-1437.
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