Trypsin from the viscera of Bogue (Boops boops): isolation and characterisation.
Fish Physiol Biochem
; 36(4): 893-902, 2010 Dec.
Article
en En
| MEDLINE
| ID: mdl-19898990
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 Km and kcat on BAPNA were 0.13 mM and 1.56 s(-1), respectively, while the catalytic efficiency kcat/Km 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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Bases de datos:
MEDLINE
Asunto principal:
Perciformes
/
Vísceras
/
Tripsina
Límite:
Animals
Idioma:
En
Revista:
Fish Physiol Biochem
Año:
2010
Tipo del documento:
Article
País de afiliación:
Túnez