title: Covalent immobilization of α-amylase on magnetic particles as catalyst for hydrolysis of high-amylose starch.
abstract: Enzyme immobilized on magnetic particles can be used as efficient recoverable biocatalysts under strong magnetic response. To enable re-use of enzyme, modified Fe3O4 particles were used as carrier to immobilize α-amylase in this paper. Firstly, the surface of Fe3O4 particles were coated with amino groups by direct using TEOS (tetraethoxysilane) followed by treatment with APTES (3-aminopropyltriethoxysilane) and then carboxylated by reacting it with succinic anhydride. In addition, the effect of the immobilization condition on enzyme activity recovery and immobilization efficiency were investigated. The results showed that the optimal immobilization occurred under following conditions: pH 5.5, 40°C, enzyme concentration of 20mgmL(-1), reaction time for 36h. Using immobilized α-amylase as biocatalyst, the optimum pH and temperature for hydrolysis were observed to be 6.5 and 60°C. The kinetics of hydrolysis reaction were studied using Michaelis-Menten equation. The affinity constant (Km) and maximum reaction rate (vmax) of magnetic particles immobilization α-amylase (MPIA) was 0.543mgmL(-1) and 1.321mgmin(-1) compared to those of 0.377mgmL(-1) and 6.859mgmin(-1) of free enzyme. After immobilization, enzymatic activity, storage stability, thermo-stability, and reusability of MPIA were found superior to those of the free one. MPIA maintained 86% enzyme activity after 30 days and maintained 78% enzyme activity after recycling six times.
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, PR China. Electronic address: tggh635@163.com.
authors.forename: Hui
authors.initials: H
authors.lastname: Guo
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Tang
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Lu
authors.initials: L
authors.lastname: Xue
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Jun-Qing
authors.initials: JQ
authors.lastname: Qian
mesh_terms: D000688:Amylose / Q000737:chemistry*; D000818:Animals; D055162:Biocatalysis; D002417:Cattle; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D013696:Temperature; D000516:alpha-Amylases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D058185:Magnetite Nanoparticles; D012997:Solvents; D000688:Amylose; D000516:alpha-Amylases
keywords: Immobilized α-amylase; Magnetic Fe(3)O(4) nanoparticles; Starch hydrolysis