Marquez-Rios Enrique, Cota-Arriola Octavio, Villalba-Villalba Ana Gloria, Ezquerra-Brauer Josafat Marina, Ocaño-Higuera Victor Manuel, Lopez-Corona Betzabe Ebenhezer, Torres-Arreola Wilfrido
1Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Blvd. Luis Encinas y Rosales s/n. Apdo. Postal 1658, C.P. 83000 Col. Centro, Hermosillo, Sonora, México.
Ingeniería Ambiental. Universidad Estatal de Sonora, Unidad Académica Hermosillo. Ley Federal del Trabajo S/N, Hermosillo, Sonora, México.
Food Sci Biotechnol. 2016 Aug 31;25(4):1011-1016. doi: 10.1007/s10068-016-0163-y. eCollection 2016.
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 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.
从巨型鱿鱼肝胰腺（HP）中纯化出糜蛋白酶，纯化倍数为2.4倍，产率为1.9%，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）估计其分子量为31 kDa，并对其进行了表征。评估了糜蛋白酶对从巨型鱿鱼的外套膜、鳍和腕部提取的胶原蛋白的作用。以琥珀酰-丙氨酰-丙氨酰-脯氨酰-苯丙氨酸对硝基苯胺（SAAPNA）为底物时，该酶在pH 7和65°C下表现出最大活性，并且使用特异性抑制剂N-对甲苯磺酰-L-苯丙氨酸氯甲基酮（TPCK）和苯甲基磺酰氟（PMSF）对其进行鉴定后发现，其残余活性分别为6%和0%。此外，在4.0至8.0的pH范围内观察到较高活性。纯化后的酶以肌肉胶原蛋白为底物时表现出中等活性。尽管还需要进一步研究，但结果表明该酶在需要酸性或弱碱性条件的情况下具有潜在应用价值。
