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  • a Triângulo Mineiro Federal University, Rua Getulio Guaritá, 159, CEP 38025 360, Uberaba, Minas Gerais, Brazil
  • | b Federal University of Alfenas, Rua Gabriel Monteiro da Silva, 700, Prédio H, sala 106D, CEP 37 130 000, Alfenas, Minas Gerais, Brazil
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Protein hydrolysis using immobilized proteases may present problems, which are mainly caused by the diffusion of macromolecular substrate. Pre-hydrolysed substrate could be a viable alternative in this process. The aim of this work was to test the casein and β-lactoglobulin hydrolysis using immobilized papain on glyoxyl-agarose, following substrate pre-treatment with immobilized trypsin. Although immobilized papain showed difficulties to degrade the milk proteins, after 10 min of trypsin pre-hydrolysis, the immobilized papain was able to achieve the maximum degree of hydrolysis in shorter time. For pre-hydrolysate casein, the immobilized papain reached 98.9% at 60 min total reaction (including the 10 min pre-digestion). As for β-lactoglobulin, immobilized papain was capable of achieving maximum levels after just 60 min of reaction, where the free enzyme only achieved 60% of its maximum. Pre-hydrolysis with trypsin appears to have worked well as a pre-treatment method to increase the proteolytic action of immobilized papain.

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