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Forming nanoparticles of α-amylase and embedding them into solid surfaces

Author:
Meridor, David, Gedanken, Aharon
Source:
Journal of Molecular Catalysis. B, Enzymatic 2013 v.90 pp. 43-48
ISSN:
1381-1177
Subject:
Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, alpha-amylase, aqueous solutions, catalytic activity, enzyme activity, glass, immobilized enzymes, leaching, maltose, nanoparticles, pH, polyethylene, potato starch, temperature, transmission electron microscopy, ultrasonics
Abstract:
In the current work nanoparticles (NPs) of α-amylase were generated in an aqueous solution using high-intensity ultrasound, and were subsequently immobilized on polyethylene (PE) films, or polycarbonate (PC) plates, or on microscope glass slides. The α-amylase NPs coated on the solid surfaces have been characterized by ESEM, TEM, FTIR, XPS and AFM. The substrates immobilized with α-amylase were used for hydrolyzing soluble potato starch to maltose. The amount of enzyme introduced in the substrates, leaching properties, and the catalytic activity of the immobilized enzyme were compared. The catalytic activity of the amylase deposited on the three solid surfaces was compared to that of the same amount of free enzyme at different pHs and temperatures. α-Amylase coated on PE showed the best catalytic activity in all the examined parameters when compared to native amylase, especially at high temperatures. When immobilized on glass, α-amylase showed better activity than the native enzyme over all pH and temperature values studied. However, the immobilization on PC did not improve the enzyme activity at any pH and any temperature compared to the free amylase. The kinetic parameters, Km and Vmax were also calculated. The amylase coated PE showed the most favorable kinetic parameters (Km=5gL−1 and Vmax=5E−07molmL−1min−1). In contrast, the anchored enzyme-PC exhibited unfavorable kinetic parameters (Km=16gL−1, Vmax=4.2E−07molmL−1min−1). The corresponding values for amylase-glass were Km=7gL−1, Vmax=1.8E−07molmL−1min−1, relative to those obtained for the free enzyme (Km=6.6gL−1, Vmax=3.3E−07molmL−1min−1).
Agid:
855203