Application of Superparamagnetic Particles in Amylase Purification

1. Abstract In recent years, many researchers pay more attention to the development of magnetic carrier technology which was used the un-magnetic particles as matrix, after modifying the surface of magnetic carrier by cross-linking agent, the target was adsorbed onto a support, and the enzyme could be purified by the separation of magnetic field. In this study, the superparamagnetic particles which were modified by cross-linking agents were used to be the support, the starch could be coated on the support, and this support would be affinity to the amylase, then, it could be used in the purification of α and β-amylase, the result showed that the support could purify the α-amylase from BSA and α-amylase complex solution, the recovery was 79.3% and the fold-purification was 36.4, the support also could purify the β-amylase from BSA and β-amylase complex solution, the recovery was 54.7% and the fold-purification was 57.8. The carrier could adsorb the target protein efficiently, so the technology may further combine with other separation technologies in industry to improve its efficiency of purification and more affinity magnetic carrier could be developed for practical application. Lane 1: Marker protein ; Lane 2: Commercial α-amylase; Lane 3: Commercial BSA; Lane 4: Commercial α-amylase and BSA mixture solution; Lane 5: Starch-SPIO purified enzyme. Lane 1 : Marker protein; Lane 2 : Commercial β-amylase; Lane 3 : Commercial BSA; Lane 4 : Commercial β-amylase and BSA mixture solution; Lane 5 : Starch-SPIO purified enzyme Figure 1. The SDS-PAGE of purified α-amylase from commercial α-amylase and BSA mixture solution by affinity superparamagnetic oxide iron particle. Figure 2. The SDS-PAGE of purified β-amylase from commercial β-amylase and BSA mixture solution by affinity superparamagnetic oxide iron particle. Application of Superparamagnetic Particles in Amylase Purification S. T. Kuo, H. H. Chen, Y. C. Hou, C. M. Jiang and M. C. Wu*National Pingtung University, TAIWAN *globalizationwu@yahoo.com.tw Result Table 1. Purification of α-amylase from commercial α-amy lase and BSA mixture solution by affinity superparamagnetic oxide iron particle. Table 2. Purification of β-amylase from commercial enzyme and BSA mixture solution by affinity superparamagnetic oxide iron particle.