Characteristics of Penicillin G Acylase Immobilized onto Iron Oxide Nanoparticles
Momen Seyed Bahman
Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran
Atyabi Seyed Mohammad
Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran
Azim Akbarzadeh
Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran
Salimi Mona
Physiology and Pharmacology Department, Pasteur Institute of Iran, Tehran 13164, Iran
Sedigeh Hatami Gigloo
Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran
Farhangi Ali
Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran
Norouzian Dariush *
Pilot Biotechnology Department, Pasteur Institute of Iran, Tehran 13164, Iran
*Author to whom correspondence should be addressed.
Abstract
Penicillin G acylase was immobilized onto iron oxide nanoparticles coated with polyethyleneimine and then cross linked with glutaraldehyde solution. The FTIR spectrum of immobilized enzyme showed peak at 1648cm-1 which can be attributed to the C=N bonds of Schiff’s base linkage formed between glutaraldehyde and amino group of penicillin G acylase. By considering the FTIR spectrum of nano particle coated with polyethyleneimine, adsorption of penicillin G acylase has taken place and then glutaraldehyde cross linked enzyme onto activated support. Catalytic properties of nano penicillin G acylase were improved upon immobilization as compared to its free counterpart. The optimal pH and temperature were determined to be 7.0, 10.0, 50 and 75ºC for free and immobilized penicillin G acylase, respectively. Thermal stabilities of both nano and free penicillin G acylase were studied .The Km value of immobilized nanozyme was calculated from Lineweaver Burck plot to be 0.23 μM while that of free penicillin G acylase was 0.28μM. In this way nano penicillin G acylase with improved catalytic properties was developed as compared to its soluble counterpart.
Keywords: Immobilized, nanopenicillin G acylase, polyethyleneimine, kinetic properties