Novel biotechnological formulations of cysteine proteases, immobilized on chitosan. Structure, stability and activity
Bromelain, papain, and ficin are studied the most for meat tenderization, but have limited application due to their short lifetime. The aim of this work is to identify the adsorption mechanisms of these cysteine proteases on chitosan to improve the enzymes' stability. It is known that immobiliz...
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Veröffentlicht in: | International journal of biological macromolecules 2021-06, Vol.180, p.161-176 |
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Sprache: | eng |
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Zusammenfassung: | Bromelain, papain, and ficin are studied the most for meat tenderization, but have limited application due to their short lifetime. The aim of this work is to identify the adsorption mechanisms of these cysteine proteases on chitosan to improve the enzymes' stability. It is known that immobilization can lead to a significant loss of enzyme activity, which we observed during the sorption of bromelain (protease activity compared to soluble enzyme is 49% for medium and 64% for high molecular weight chitosan), papain (34 and 28% respectively) and ficin (69 and 70% respectively). Immobilization on the chitosan matrix leads to a partial destruction of protein helical structure (from 5 to 19%). Using computer modelling, we have shown that the sorption of cysteine proteases on chitosan is carried out by molecule regions located on the border of domains L and R, including active cites of the enzymes, which explains the decrease in their catalytic activity upon immobilization. The immobilization on chitosan does not shift the optimal range of pH (7.5) and temperature values (60 °C for bromelain and papain, 37–60 °C for ficin), but significantly increases the stability of biocatalysts (from 5.8 times for bromelain to 7.6 times for papain).
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•Chitosan matrix blocks in part active cites of adsorbed cysteine proteases.•Immobilization on chitosan leads to partial destruction of enzyme helical structure.•Blocking and alteration of structure decrease enzymatic activity.•Immobilization does not shift pH and temperature optima of enzymes.•Immobilization significantly increases stability of biocatalysts. |
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ISSN: | 0141-8130 1879-0003 |
DOI: | 10.1016/j.ijbiomac.2021.03.016 |