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KMID : 0545120160260050829
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Journal of Microbiology and Biotechnology
2016 Volume.26 No. 5 p.829 ~ p.836
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Covalent Immobilization of Penicillin G Acylase onto Fe3O4@Chitosan Magnetic Nanoparticles
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Ling Xiao Min
Wang Xiang Yu
Ma Ping
Yang Yi
Qin Jie Mei
Zhang Xue Jun
Zhang Ye Wang
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Abstract
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Penicillin G acylase (PGA) was immobilized on magnetic Fe3O4@chitosan nanoparticles through the Schiff base reaction. The immobilization conditions were optimized as follows: enzyme/support 8.8 mg/g, pH 6.0, time 40 min, and temperature 25oC. Under these conditions, a high immobilization efficiency of 75% and a protein loading of 6.2 mg/g-support were obtained. Broader working pH and higher thermostability were achieved by the immobilization. In addition, the immobilized PGA retained 75% initial activity after ten cycles. Kinetic parameters Vmax and Km of the free and immobilized PGAs were determined as 0.91 mmol/min and 0.53 mmol/min, and 0.68 mM and 1.19 mM, respectively. Synthesis of amoxicillin with the immobilized PGA was carried out in 40% ethylene glycol at 25°C and a conversion of 72% was obtained. These results showed that the immobilization of PGA onto magnetic chitosan nanoparticles is an efficient and simple way for preparation of stable PGA.
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KEYWORD
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Chitosan, Immobilization, Magnetic nanoparticles, penicillin G acylase, Amoxicillin
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