Selective Immobilization of His-Tagged Enzyme on Ni-Chelated Ion Exchange Resin and Its Application in Protein Purification

Abstract

Ion exchange resins are suitable as carriers for immobilized enzymes because of their stable physicochemical properties, appropriate particle size and pore structure, and lower loss in continuous operation. In this paper, we report the application of the Ni-chelated ion exchange resin in the immobilization of His-tagged enzyme and protein purification. Acrylic weak acid cation exchange resin (D113H) was selected from four cationic macroporous resins that could chelate the transition metal ion Ni. The maximum adsorption capacity of Ni was ~198 mg/g. Phosphomannose isomerase (PMI) can be successfully immobilized on Ni-chelated D113H from crude enzyme solution through chelation of transition metal ions with the His-tag on the enzyme. The maximum amount of immobilized PMI on the resin was ~143 mg/g. Notably, the immobilized enzyme showed excellent reusability and maintained 92% of its initial activity with 10 cycles of catalytic reaction. In addition, PMI was successfully purified using an affinity chromatography column prepared by Ni-chelated D113H, which showed the potential for the immobilization and purification process to be realized in one step.

One-Step Purification and Immobilization of Glycosyltransferase with Zn–Ni MOF for the Synthesis of Rare Ginsenoside Rh2

“…19 Transition metal ions (Cu 2+ , Zn 2+ , Ni 2+ , and Co 2+ ) can form coordination bonds with imidazole residues on His, which enables their firm adsorption onto the carrier. 20 Moreover, His-tags have minimal effects on protein solubility, physical properties, or structure. Compared with random immobilization, directional immobilization through affinity tags results in higher enzyme activity and improved thermal and operational stabilities.…”

“…His-tags are most commonly used due to their ease of expression and cost effectiveness in Escherichia coli host bacteria . Transition metal ions (Cu 2+ , Zn 2+ , Ni 2+ , and Co 2+ ) can form coordination bonds with imidazole residues on His, which enables their firm adsorption onto the carrier . Moreover, His-tags have minimal effects on protein solubility, physical properties, or structure.…”

Magnetic resin composites for the enrichment of proteins, peptides and phosphopeptides

“…It is well known that resins can be employed for protein adsorption due to their stable physicochemical properties, appropriate particle size and pore structure. 12 Ion-exchange resins with acidic or basic functional groups possess the advantages of low cost, good mechanical strength, and stable physical and chemical properties, and also have broad application prospects. 13 The electrostatic interaction between the charge on the proteins and the charged groups of the ion exchange resins also facilitates protein purification.…”