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Purification and characterization of cysteine proteinase inhibitors from crucian carp Carassius auratus eggs

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  • Chemistry and Biochemistry
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Abstract

Two cystatins (cst-I and cst-II) were purified from crucian carp eggs by acidification and subsequent ion exchange and molecular sieve chromatography. The molecular masses of cst-I and cst-II analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis were 11.9 and 14.4 kDa, respectively, under reducing conditions and 13.5 and 12.7 kDa, respectively, under non-reducing conditions. The cst-I and cst-II molecules were stable after 30 min of incubation at 60 and 50°C, respectively. There was no significant loss in the inhibitory activity of either cst in the pH range 4–11. These two cystatins were able to affect the proteolysis of papain, cathepsin L, and bromelain, but they were unable to inhibit cathepsin B and trypsin. The partial N-terminal amino acid sequences of both cst inhibitors were homologous and that of cst-I was recognized as NH2-AGIPGGLVDADINDADVQ. This latter fragment shared 88.9% identity to common carp cystatin and 44.4–55.6% to cystatins of other aquatic animals. Based on these results, we conclude that the two cst inhibitors are members of family II cystatin.

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Acknowledgments

This study was supported by the National Science Council, ROC, under Grant Nr. NSC 94-2313-B-041-009, and Chia-Nan University of Pharmacy & Science, under Grant Nr. CN9731.

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Authors and Affiliations

  1. Department of Hotel and Restaurant Management, Chia-Nan University of Pharmacy and Science, 60 Erh-Jen Rd., Sect. 1, Jen-Te, Tainan, 71710, Taiwan

    Shinn-Shuenn Tzeng, Hui-Chun Wu & Wen-Chieh Sung

  2. Department of Biotechnology, Chia-Nan University of Pharmacy and Science, 60 Erh-Jen Rd., Sect. 1, Jen-Te, Tainan, 71710, Taiwan

    Chien-An Liu

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  1. Shinn-Shuenn Tzeng
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Correspondence to Shinn-Shuenn Tzeng.

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Tzeng, SS., Wu, HC., Sung, WC. et al. Purification and characterization of cysteine proteinase inhibitors from crucian carp Carassius auratus eggs. Fish Sci 75, 1453–1460 (2009). https://doi.org/10.1007/s12562-009-0170-5

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  • DOI: https://doi.org/10.1007/s12562-009-0170-5

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