Preparation of an enzyme/inorganic nanosheet/magnetic bead complex and its enzymatic activity

Abstract

A complex composed of enzyme (horseradish peroxidase, HRP), layered titanate (TiO x ), and magnetic beads (MB) was prepared, and the enzymatic activity of the complex and the repeatable use for the enzyme in the complex were investigated. The HRP/TiO x /MB complex was formed in acetate buffer (pH 4) by electrostatic interaction among the components where HRP and MB have positive charge, whereas TiO x is negatively charged. The resultant complex was sufficiently stable in an aqueous solution, and HRP immobilized on the layered titanate in the complex kept a certain enzymatic activity in which typical enzymatic relationship was observed between substrate concentration and initial reaction rate. By giving a magnetic field using commercial magnets, the HRP/TiO x / MB complex was easily and rapidly recovered, and the recovered complex was able to be used for the next enzymatic reactions. The enzyme/inorganic nanosheet/MB complex proposed in this study is expected to be applied as a new tool in the bioengineering and nanobiotechnological fields.

Antioxidant Materials Based on 2D Nanostructures: A Review on Recent Progresses

“…Stable suspensions were obtained by immobilizing HRP and magnetic beads on positively charged layered titania in slightly acidic samples [111]. Although steric hindrance and possibly slight The same TNS was used to immobilize SOD enzyme [61].…”

“…Stable suspensions were obtained by immobilizing HRP and magnetic beads on positively charged layered titania in slightly acidic samples [111]. Although steric hindrance and possibly slight conformational changes led to reduced HRP activity, the magnetic composite was separable with strong magnets at the end of catalytic cycles.…”

Enhanced catalytic activity of enzymes interacting with nanometric titanate nanosheets

“…One advantage of immobilized enzymes is that tiny enzyme molecules are easily recovered from a reaction system and can then be reused many times. 3 In general, the stability of an enzyme is reinforced in proportion to the binding energy to the solid support. 4 Although thermal or chemical durability of immobilized enzymes is surely reinforced, an apparent enzymatic activity at an ambient atmosphere is reduced because the binding to a solid support prohibits Brownian motion of enzyme molecules in a solvent, causing deterioration in accessibility to external substrate molecules.…”

Oxidation of Organic Compounds by Magnetic Biocatalyst

“…Одним из перспективных применений является использование их в качестве носителей для иммобилизованных ферментов. Так например, фермент класса оксидоредуктаз Нorseradish peroxidase (HRP) был иммобилизован на магнит-модифицированный полианилин активированный глутаровым диальдегидом [8], на магнитные железотитановые пластинки [1], на магнитные наночастицы с использованием 3-аминопропилтриэтоксисилана (3-и глутарового диальдегида в качестве сшивающего агента [6], на магнитные наночастицы с использованием хитозана и полиэтиленмалеиновой кислоты [9], на магнитные поли(глицидилметакрилат-со-метилметакрилат)ные гранулы активированные с помощью глутарового диальдегида [10], на магнитные наночастицы методом адсорбции [11]…”