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Purification and characterization of a cysteine-like protease from the body wall of the sea cucumber Stichopus japonicus

Fish Physiology and Biochemistry volume 33, pages 181–188 (2007)Cite this article

Abstract

The sea cucumber (Stichopus japonicus) is able to undergo autolysis in response to a variety of environmental and mechanical cues. Within the framework of a long-term study of this phenomenon we have purified a protease from the body wall of the sea cucumber by means of ion-exchange chromatography with DE-52 cellulose and gel filtration chromatography with Sephadex G-100. The final enzyme preparation was nearly homogeneous on polyacrylamide gel electrophoresis, and its molecular weight was estimated to be approximately 35.5 kDa. The purified enzyme exhibited a maximum activity for the hydrolysis of casein at pH 7.0 and 50°C and a remarkable stability at pH 4.0–7.0 and 40–60°C. Based on the inhibition and activation profiles obtained with numerous specific protease inhibitors and an activator, the protease purified from the body wall of the sea cucumber was defined to be a cysteine-like protease.

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Acknowledgements

We thank Prof. Murata Yoshiyuki (Department of Biological Resources Chemistry, Faculty of Agriculture, Okayama University) for supplying some of the agents and for providing us with useful suggestions. This work was financially supported by the National Natural Science Foundation of China (NSFC; No. 30571449), Chinese International Corporation Project (No. 2006DFA32580), and National Basic Research Program of China (973 Program; No. 2006CB708210).

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Authors and Affiliations

  1. College of Bio & Food Technology, Dalian Institute of Light Industry, Qinggongyuan No.1, Ganjingzi District, Dalian, 116034, P.R. China

    H. Qi, X. P. Dong, L. N. Cong, Y. Gao, L. Liu & B. W. Zhu

  2. Department of Biological Resources Chemistry, Faculty of Agriculture, Okayama University, Okayama, 700-8530, Japan

    T. Mikiro

Authors
  1. H. Qi
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  2. X. P. Dong
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  3. L. N. Cong
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  4. Y. Gao
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  5. L. Liu
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  6. T. Mikiro
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  7. B. W. Zhu
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Correspondence to B. W. Zhu.

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Qi, H., Dong, X.P., Cong, L.N. et al. Purification and characterization of a cysteine-like protease from the body wall of the sea cucumber Stichopus japonicus . Fish Physiol Biochem 33, 181–188 (2007). https://doi.org/10.1007/s10695-007-9129-6

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  • DOI: https://doi.org/10.1007/s10695-007-9129-6

Keywords

  • Characterization
  • Proteases
  • Purification
  • Sea cucumber (Stichopus japonicus)

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