Chymotrypsin isolation from jumbo squid (<Emphasis Type="Italic">Dosidicus gigas</Emphasis>) hepatopancreas: Partial characterization and effect on muscle collagen |
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Authors: | Enrique Marquez-Rios Octavio Cota-Arriola Ana Gloria Villalba-Villalba Josafat Marina Ezquerra-Brauer Victor Manuel Ocaño-Higuera Betzabe Ebenhezer Lopez-Corona Wilfrido Torres-Arreola |
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Affiliation: | 1.Departamento de Investigación y Posgrado en Alimentos,Universidad de Sonora,Hermosillo, Sonora,México;2.Ingeniería Ambiental. Universidad Estatal de Sonora,Unidad Académica Hermosillo. Ley Federal del Trabajo S/N,Hermosillo, Sonora,México;3.Departamento de Ciencias Químico Biológicas,Universidad de Sonora,Hermosillo, Sonora,México |
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Abstract: | Chymotrypsin was purified from jumbo squid hepatopancreas (HP) with 2.4-fold and yield 1.9%, and characterized with a molecular weight of 31 kDa, as estimated by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). Chymotrypsin effect over collagen extracted from the mantle, fins and arms of the jumbo squid was evaluated. The enzyme exhibited the maximum activity at pH 7 and 65°C using Suc-Ala-Ala-Pro-Phe-p-nitroanilide (SAAPNA) as a substrate and it was identified using the specific inhibitors N-tosyl-L-phenylalaninechloromethyl ketone (TPCK) and phenyl methyl sulfonyl fluoride (PMSF), showing residual activities of 6% and 0%, respectively. Furthermore, high activity was observed in the pH range of 4.0 to 8.0. Purified enzyme showed a moderate in vitro activity using muscle collagen as a substrate. Although further research is needed, the results suggest that the enzyme has a potential application where acidic or slightly alkaline conditions are needed. |
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