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TIP Revista Especializada en Ciencias Químico-Biológicas

2019, Number 1

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TIP Rev Esp Cienc Quim Biol 2019; 22 (1)

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

Osuna-Amarillas PS, Rouzaud-Sandez O, Higuera-Barraza OA, Arias-Moscoso JL, López-Mata MA, Campos-García JC, Valdez-Melchor RG

Language: Spanish
References: 23
Page: 1-10
PDF size: 448.96 Kb.


ABSTRACT

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.


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