Purification and Characterization of Cathepsin B from the Muscle of Horse Mackerel Trachurus japonicus
AbstractAn endogenous protease in fish muscle, cathepsin B, was partially purified and characterized from horse mackerel meat. On SDS-PAGE of the purified enzyme under reducing conditions, main protein bands were detected at 28 and 6 kDa and their respective N-terminal sequences showed high homology to heavy and light chains of cathepsin B from other species. This suggested that horse mackerel cathepsin B formed two-chain forms, similar to mammalian cathepsin Bs. Optimum pH and temperature of the enzyme were 5.0 and 50 °C, respectively. A partial cDNA encoding the amino acid sequence of 215 residues for horse mackerel cathepsin B was obtained by RT-PCR and cloned. The deduced amino acid sequence contains a part of light and heavy chains of cathepsin B. The active sites and an N-glycosylation site were conserved across species. We also confirmed that the modori phenomenon was avoided by CA-074, a specific inhibitor for cathepsin B. Therefore, our results suggest that natural cysteine protease inhibitor(s), such as oryzacystatin derived from rice, can apply to thermal-gel processing of horse mackerel to avoid the modori phenomenon. Meanwhile, this endogenous protease may be used for food processing, such as weaning meal and food for the elderly. View Full-Text
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Yoshida, A.; Ohta, M.; Kuwahara, K.; Cao, M.-J.; Hara, K.; Osatomi, K. Purification and Characterization of Cathepsin B from the Muscle of Horse Mackerel Trachurus japonicus. Mar. Drugs 2015, 13, 6550-6565.
Yoshida A, Ohta M, Kuwahara K, Cao M-J, Hara K, Osatomi K. Purification and Characterization of Cathepsin B from the Muscle of Horse Mackerel Trachurus japonicus. Marine Drugs. 2015; 13(11):6550-6565.Chicago/Turabian Style
Yoshida, Asami; Ohta, Megumi; Kuwahara, Koichi; Cao, Min-Jie; Hara, Kenji; Osatomi, Kiyoshi. 2015. "Purification and Characterization of Cathepsin B from the Muscle of Horse Mackerel Trachurus japonicus." Mar. Drugs 13, no. 11: 6550-6565.