title: Structural and catalytic properties of immobilized α-amylase from Laceyella sacchari TSI-2.
abstract: One of the approaches to address the issues of the cost of production, recovery and reusability of the extremozymes can be immobilization. In this report, we describe immobilization of an α-amylase from Laceyella sacchari TSI-2 and characterization of the immobilized enzyme. The enzyme was immobilized on 6 different matrices using entrapment, ionic binding and surface adsorption. The DEAE cellulose with glutaraldehyde crosslinking appeared most effective for the immobilization with high operational stability. While the temperature optima and thermal stability of the immobilized α-amylase shifted from 60 to 70°C with increased half-life, the pH optima remain unaltered while pH stability was shifted from 6 to 7. The stability of the immobilized enzyme improved in solvents. The enzyme catalysis in surfactants enhanced, while the Km and Vmax were reduced after immobilization. The structural features of the immobilized enzyme as probed by FT-IR established the role of aliphatic amines, esters and alkenes in immobilization. The starch hydrolysis efficiency of the immobilized enzyme was 15.55%. The immobilized enzyme in various detergents was highly efficient in removing the starch stain from cotton cloth. Taken together, the α-amylase turned more stable after immobilization and can be a favored choice for applications.
authors.affiliation: UGC-CAS Department of Biosciences, Saurashtra University, Rajkot 360 005 Gujarat, India.
authors.forename: Rushit J
authors.initials: RJ
authors.lastname: Shukla
authors.affiliation: UGC-CAS Department of Biosciences, Saurashtra University, Rajkot 360 005 Gujarat, India. Electronic address: satyapsingh@yahoo.com.
authors.forename: Satya P
authors.initials: SP
authors.lastname: Singh
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D002384:Catalysis; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D011994:Recombinant Proteins; D012997:Solvents; D017550:Spectroscopy, Fourier Transform Infrared; D013329:Structure-Activity Relationship; D013501:Surface-Active Agents / Q000737:chemistry; D013696:Temperature; D000516:alpha-Amylases / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D012997:Solvents; D013501:Surface-Active Agents; D000516:alpha-Amylases
keywords: FT-IR; Immobilization; Laceyella sacchari; Saccharification; Thermoactinomycete; α-Amylase