Hydrophobic interaction chromatography as separation method of alkaline proteases from viscera of Scomberomorus sierra

• Main Authors: Pablo Sergio Osuna-Amarillas, Ofelia Rouzaud-Sandez, Odilia Azucena Higuera-Barraza, Joe Luis Arias-Moscoso, Marco Antonio López-Mata, Julio César Campos-García, Ramón Gertrudis Valdez-Melchor
• Format: Article
• Language: English
• Published: Universidad Nacional Autónoma de México 2019-07-01
• Series: TIP Revista Especializada en Ciencias Químico-Biológicas
• Subjects: chymotrypsin; hydrophobic interaction chromatography; scomberomorus sierra; trypsin
• Online Access: http://tip.zaragoza.unam.mx/index.php/tip/article/view/183

This study focused on recovering alkaline proteases from the viscera of Scomberomorus sierra through hydrophobic interaction chromatography. Three alkaline proteases were partially separated using this chromatographic technique; two of them, with molecular weights of 19 and 31 kDa, were identified as trypsin-like enzymes according to inhibition assays. The 31 kDa alkaline protease, the only isolated enzyme, was purified under following chromatographic conditions: ammonium sulfate 13% (w/v) and ethylene glycol 27% (w/v); this enzyme showed maximum activity at pH 9 – 10 and 50 – 60 °C and was strongly inhibited by soybean trypsin inhibitor (SBTI) and porcine trypsin inhibitor (TPI). A third alkaline protease with molecular weight of 20 kDa was partially separated and inhibited by tosyl phenylalanyl chloromethyl ketone (TPCK), showing optimum activity at pH 9 – 11 and 60 °C. These results show that the viscera of Scomberomorus sierra may be useful as source of proteases.