Construction of novel curdlan-based and Ca2+-chelated magnetic microspheres (CCMM) for efficient protein purification and oriented immobilization.

Enzyme and Microbial Technology
Yifeng ZhangYingfeng An

Abstract

In this study, curdlan-based and calcium ion (Ca2+)-chelated magnetic microspheres (CCMM) were prepared for protein purification and oriented immobilization. Additional purification steps before immobilization were not required. CCMM samples were produced by reverse embedding of Fe3O4 nanoparticles with curdlan and chelated with Ca2+ in the presence of iminodiacetic acid. The β-xylanase XynII from Trichoderma reesei QM6a was used to investigate the efficiency of CCMM preparation. The resulting CCMM-XynII was found to be very stable, showing 82 % and 60 % of initial activities after storage for 35 days and after being assayed ten times, respectively. In addition, the CCMM-XynII showed higher stabilities in the presence of organic solvents and multiple chemicals than the free XynII, suggesting that the CCMM-XynII could be efficient for applications requiring the presence of organic solvents. In addition, CCMM may be more suitable than commercially available Ni-NTA for purification of proteins intolerant of Ni2+.

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