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

作者：

Qilong Tang, Wenli Wang, Lujia Zhang, Yuan Liu

期刊：

Journal of Food Measurement and Characterization

摘要：

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.

关键词：

 Takifugu rubripes; Cathepsin L;  Biochemical characterization;  Flavor peptides

Fig. 1 The comparison of recombinant TrCL and original TrCL after sequencing