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Purification and characterization of stomach protease from the turbot (Scophthalmus maximus L.)

Fish Physiology and Biochemistry volume 32, pages 179–188 (2006)Cite this article

Abstract

Proteolytic activity in the different parts of the digestive tract of the turbot (Scophthalmus maximus L.) were studied in this work. One pure protease was isolated from turbot stomach and its behavior was studied. Results showed the optimum pH for proteases in the different parts of the digestive tract of the turbot were pH 2.0 for the stomach, pH 8.0 for the pylorus cecum, pH 8.0 for the foregut, pH 8.5 for the midgut, and pH 8.0 for the hindgut. The activity of proteases in the different parts of the digestive tract were in the sequence pylorus cecum protease > stomach protease > foregut protease > midgut protease > hindgut protease. The stomach protease was purified by ammonium sulfate precipitation and column chromatography on DEAE-Sepharose F.F. and Sephadex G-100. The purified enzyme gave a single band in SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Its molecular weight was found to be approximately 42,000 Da. The enzyme is stable at pH 1.0–9.0 and at temperatures below 40°C. Its activity was maximum at pH 2.0 and 40°C. When reaction time was prolonged the optimum temperature of the enzyme tended to decline. The enzyme was activated by Mn2+ and Cu2+ and inactivated by Fe3+. It was fully inhibited by pepstatin and partially inhibited by PMSF, TPCK, PCMB, and NBS. These results imply the enzyme is a pepsin.

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Acknowledgements

This work was supported by Science Developing project “Studies on the Digestion Enzyme in Turbot” of Science and Technology Bureau, Qingdao municipality, Shandong Province, China.

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Authors and Affiliations

  1. Yellow Sea Fisheries Research Institute, Academic of Fishery Science of China, No. 106, Nan Jing Road, Qingdao, Shandong Province, 266071, P.R. China

    Hai-ying Wang, Yue-jun Wang, Qing-yin Wang & Mi Sun

  2. Department of Food Science and Technology, Ocean University of China, No. 5, Yu Shan Road, Qingdao, Shandong Province, 266003, P.R. China

    Chang-hu Xue

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  1. Hai-ying Wang
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  2. Yue-jun Wang
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  5. Mi Sun
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Correspondence to Mi Sun.

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Wang, Hy., Wang, Yj., Wang, Qy. et al. Purification and characterization of stomach protease from the turbot (Scophthalmus maximus L.). Fish Physiol Biochem 32, 179–188 (2006). https://doi.org/10.1007/s10695-006-0010-9

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  • DOI: https://doi.org/10.1007/s10695-006-0010-9

Keywords

  • Characterization
  • Purification
  • Stomach protease
  • Turbot (Scophthalmus maximus L.)

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