Enhancing purification of α-amylase by superparamagnetic complex with alginate/chitosan/β-cyclodextrin/TPP

Abstract

The main aim of this study was to synthesize the superparamagnetic nanoparticles coated by alginate/chitosan/β-cyclodextrin to purify α-amylase. Isolated bacteria were identified by morphological, biochemical and taxonomic molecular studies. FTIR- spectrometer, VSM, X-ray instruments and Malvern Zetasizer were used to characterize nanoparticles characteristics. The morphological structures and the elemental composition of the nanoparticles were studied by using FESEM and EDS, respectively. The molecular weight of enzyme was determined using SDS-PAGE, and the enzyme activity detected by zymographic analysis. FTIR studies showed the presence of Fe–O–Fe in the Fe₃O₄ and verified the interaction between chitosan, β-cyclodextrin and alginate. The saturation magnetization for superparamagnetic and coated superparamagnetic nanoparticles was indicated 39 and 1.9 emu g⁻¹, respectively. The maximum intensity of the XRD peak indicated the presence of the Fe₃O₄. FESEM and EDS analysis showed that the nanoparticles were regular and spherical in shape and corresponded to the Fe and O elements. Enzyme purification by synthesized nanoparticles was achieved 13.84 U mg⁻¹; purification fold of 3.50. The molecular weight of α-amylase was about 22 kDa. The highest activity of α-amylase was observed at 70 °C, pH 9.3 and Ca²⁺-independent. As a conclusion, the coated superparamagnetic nanoparticles showed more applications in enzyme purification comparing to the conventional methods.

Keywords:

• Superparamagnetic
• purification
• amylase
• nanoparticles

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The work was a part of the MSC thesis and has been kindly supported by the Kerman University of Medical Sciences (KMU) of Iran.