Enhanced thermal and ultrasonic stability of a fungal protease encapsulated within biomimetically generated silicate nanospheres.
BACKGROUND
Dendrimers are highly branched synthetic macromolecules with a globular shape. They have been successfully used for generation of nanospheres at mild conditions via biomimetic silicification. Encapsulation of enzyme molecules within these nanospheres during their synthesis is a promising method for rapid and efficient entrapment of several enzymes. However, encapsulation of proteolytic enzymes has been rarely done via biomimetic silicification. As well, the operational stability of encapsulated enzyme has not been systematically reported.


METHODS
A proteolytic enzyme, either alpha-Chymotrypsin or a fungal protease from Aspergilus Oryzea was encapsulated along with iron oxide nanoparticles within particles yielded via biomimetic silicification of different generations of polyamidoamine (PAMAM) dendrimers. Stability of encapsulated enzyme was compared to that of free enzyme during storage at room temperature. As well, their thermal and ultrasonic stabilities were measured. Scanning electron microscopy, transmission electron microscopy and optical microscopy were used to investigate the morphology of nanospheres.


RESULTS
Determination of encapsulation efficiency revealed that approximately 85% of fungal protease with concentration 1.4mg mL(-1) stock solution was immobilized within particles yielded by generation 0. Based on microscopic images the generated particles interconnected with each other and had spherical morphologies independent of generation. Kinetic analysis of encapsulated fungal protease demonstrated that Mechaelis-Menten constant (K(m)) slightly increased.


CONCLUSIONS
PAMAM dendrimer generation 0 could be effectively used for rapid encapsulation of a fungal protease from Aspegilus Oryzae.


CONCLUSIONS
Encapsulation significantly enhances the thermal and ultrasonic stabilities of enzymes, suggesting a range of diverse applications for them.
Madadlou
Ashkan
A
Department of Food Science & Engineering, Faculty of Technology, Campus of Agriculture & Natural Resources, University of Tehran, Karadj, Iran. ashkan.madadlou@gmail.com
Iacopino
Daniela
D
Sheehan
David
D
Emam-Djomeh
Zahra
Z
Mousavi
Mohammad E
ME
D001236:Aspergillus oryzae / Q000201:enzymology*; D002214:Capsules; D002918:Chymotrypsin / Q000737:chemistry / Q000648:ultrastructure; D050091:Dendrimers / Q000737:chemistry*; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry*; D007700:Kinetics; D008970:Molecular Weight; D053758:Nanoparticles; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D011487:Protein Conformation; D017640:Silicates / Q000737:chemistry; D014465:Ultrasonics
D002214:Capsules; D050091:Dendrimers; D005656:Fungal Proteins; C104700:PAMAM Starburst; D017640:Silicates; D010447:Peptide Hydrolases; D002918:Chymotrypsin