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RevistaPROCESS BIOCHEMISTRY
Año2017
Volumen58
Páginas98-104
Internacional

Stabilization of ficin extract by immobilization on glyoxyl-agarose. Preliminary characterization of the biocatalyst performance in hydrolysis of proteins

Autores:Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
El-Hocine Siar a,b, Hadjer Zaaka,b,c, Jakub F. Korneckia, Mohammed Nasreddine Zidouneb, Oveimar Barbosad, Roberto Fernandez-Lafuente a,*
 
Departamento de Biocatálisis.  Instituto de Catálisis-CSIC, Campus UAM-CSIC Madrid, spain.
b Equipe TEPA, Laboratoire LNTA, INATAA, Université des Frères Mentouri Constantine1, 25000, Constantine, Algeria
Food Biotechnology Division, Biotechnology Research Center (CRBt), Algeria
c  Applied Microbiology in Environment Laboratory, Science Department, Ibn Khaldoun University, Algeria.
dDepartamento de Química, Facultad de Ciencias. Universidad del Tolima, Ibagué, Colombia.



A protein extract containing ficin was immobilized on glyoxyl-agarose at pH 10 and 25ºC. The free enzyme remained fully active after 24 h at pH 10. However the enzyme immobilized on the support retained only 30% of the activity after this time using a small substrate. After checking the stability of ficin preparations obtained after different enzyme-support multi-interaction times, it was found that it reached a maximum at 3 h (40 folds more stable than the free enzyme at pH 5). The immobilized enzyme was active in a wide range of pH (e.g., retained double activity at pH 10 than the free enzyme) and temperatures (e.g., at 80ºC retained 3 folds more activity than the free enzyme). The activity versus casein almost matched the results using the small substrate (60%) at 55ºC. However in the presence of 2 M of urea, it became 3 times more active than the free enzyme. The immobilized enzyme could be reused 5 cycles at 55ºC without losing activity.
Palabras clave:protease stabilization, protease immobilization, improved enzyme performance, ficin, glyoxyl agarose, protein hydrolysis
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