Abstract
Trypsin from the viscera of Bogue (Boops boops) was purified to homogeneity by precipitation with ammonium sulphate, Sephadex G-100 gel filtration and Mono Q-Sepharose anion exchange chromatography, with an 8.5-fold increase in specific activity and 36% recovery. The molecular weight of the purified enzyme was estimated to be 23 kDa by SDS–PAGE and size exclusion chromatography. The purified trypsin appeared as a single band on native-PAGE and zymography staining. The purified enzyme showed esterase-specific activity on N-α-benzoyl-l-arginine ethyl ester (BAEE) and amidase activity on N-α-benzoyl-dl-arginine-p-nitroanilide (BAPNA). The optimum pH and temperature for the enzyme activity, after 10 min incubation, were pH 9.0 and 55°C, respectively, using BAPNA as a substrate. The trypsin kinetic constants K m and k cat on BAPNA were 0.13 mM and 1.56 s−1, respectively, while the catalytic efficiency k cat /K m was 12 s−1 mM−1. Biochemical characterisation of B. boops trypsin showed that this enzyme can be used as a possible biotechnological tool in the fish processing and food industries.
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This work was funded by «Ministry of Higher Education, Scientific Research and Technology-Tunisia».
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Barkia, A., Bougatef, A., Nasri, R. et al. Trypsin from the viscera of Bogue (Boops boops): isolation and characterisation. Fish Physiol Biochem 36, 893–902 (2010). https://doi.org/10.1007/s10695-009-9365-z
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DOI: https://doi.org/10.1007/s10695-009-9365-z