Resumo

OSUNA-AMARILLAS, Pablo Sergio et al. Hydrophobic interaction chromatography as separation method of alkaline proteases from viscera of Scomberomorus sierra. TIP [online]. 2019, vol.22, e183.  Epub 04-Mar-2020. ISSN 1405-888X.  https://doi.org/10.22201/fesz.23958723e.2019.0.183.

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.

Palavras-chave : chymotrypsin; hydrophobic interaction chromatography; Scomberomorus sierra; trypsin.

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