Cloning, purification and biochemical characterization of recombinant Cathepsin L from Takifugu rubripes and its role in taste formation

Abstract

In recent years Takifugu rubripes has been increasingly popular among people base on its good taste. Cathepsin L was related to the formation of amino acids and small peptides. This study described the biochemical characterization of Cathepsin L and its role in the taste formation of Takifugu rubripes. Cathepsin L gene from Takifugu rubripes tissues was cloned successfully, and Cathepsin L with molecular masses of 46 kDa was isolated and enzymatically characterized. This enzyme reached its highest activity at 40 °C and pH 5.5. Cu2+ and Mn2+ ions reduced the activity of Cathepsin L, but Fe3+ and Ca2+ ions prominently increased its activity when the final concentrations of metal ions were 1 mM and 5 mM, respectively. The addition of phenylmethanesulfonyl fluoride gradually decreased the enzyme activity over 0.5 mM. 200–3000 KDa fraction from Takifugu rubripes muscle after Cathepsin L hydrolysis showed the stronger kokumi and umami taste, especially kokumi. In this study, the recombinant Cathepsin L from Takifugu rubripes could play a role in flavor formation and be applied in flavor studies through the degradation of animal proteins.

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Acknowledgements

This work was funded by The National Key R&D Program of China (2016YFD0400803, 2016YFD0401501) and National Natural Science Foundation of China (Grant Nos. 31622042, 31371790, 31271900).

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Correspondence to Yuan Liu.

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Tang, Q., Wang, W., Zhang, L. et al. Cloning, purification and biochemical characterization of recombinant Cathepsin L from Takifugu rubripes and its role in taste formation. Food Measure 14, 485–491 (2020). https://doi.org/10.1007/s11694-019-00122-7

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Keywords

  • Takifugu rubripes
  • Cathepsin L
  • Biochemical characterization
  • Flavor peptides