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RevistaINTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Año2019
Volumen130
Páginas564-572
Internacional

Improved features of a highly stable protease from Penaeus vannamei by immobilization on glutaraldehyde activated graphene oxide nanosheets

Autores:Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
Neda Ranjbari, Mozhgan Razzaghi, Roberto Fernandez-Lafuente, Fozieh Shojaei, Mohammad
Satari, Ahmad Homaei
aDepartment of Marine Biology, Faculty of Marine Science and Technology, University of
Hormozgan, Bandar Abbas, Iran
bDepartamento de Biocatálisis. ICP-CSIC, Campus UAM-CSIC Madrid, Spain
cDepartment of Biology, Faculty of Sciences, Malayer University, Malayer, Iran

In this study,we report the synthesis of graphene oxide nanosheets (GON) by a modified Hummers method. Then, a
protease purified from the Penaeus vannamei shrimp was immobilized on the GON activated with glutaraldehyde.
Several techniques such as SEM, DLS and FTIR were applied to characterize the different nano-structures at the
different levels. The immobilization of the protease on the GON activated with glutaraldehyde did not affect the
optimum pH, but significantly improved thermal stability and stability at extreme pH values, as well as activity at 90ºC.
After 24 h of incubation at 90 °C,the free enzyme retained less than 10% of the activity, while the immobilized enzyme
kept more than 90% of its original activity. The apparent Km and Vmax for Penaeus vannamei protease remained
fairly similar after immobilization, a very relevant data considering the large size of the substrate (casein).In the
hydrolysis of casein at 70ºC and in the presence of 2 M urea, the immobilized enzyme exhibited a higher activity than
the free enzyme. The results indicate that the immobilization of the enzyme Penaeus vannamei protease on GON
activated with glutaraldehyde increases its already high stability against environmental stresses and makes it suitable
for biotechnological and industrial applications.
Palabras clave:Enzyme stabilization, Graphene oxide nanosheets; Penaeus vannamei protease; enzyme immobilization, glutaraldehyde.
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