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Effects of Pulsed Electric Field (PEF) Treatment on Enhancing Activity and Conformation of α-Amylase

Journal 2016 · IF 1.029 · 35(2): 154-162 · 发表于 2016 年 2 月 20 日
作者: Tian Mei-ling,Fang Ting,Du Mu-ying,Zhang Fu-sheng
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摘要

To explore an efficient, safe, and speedy application of pulsed electric field (PEF) technology for enzymatic modification, effects of PEF treatment on the enzymatic activity, property and kinetic parameters of α-amylase were investigated. Conformational transitions were also studied with the aid of circular dichroism (CD) and fluorescence spectra. The maximum enzymatic activity of α-amylase was obtained under 15 kV/cm electric field intensity and 100 mL/min flow velocity PEF treatment, in which the enzymatic activity increased by 22.13 ± 1.14 % compared with control. The activation effect could last for 18 h at 4 °C. PEF treatment could widen the range of optimum temperature for α-amylase, however, it barely exerted any effect on the optimum pH. On the other hand, α-amylase treated by PEF showed an increase of V max , t 1/2 and Δ G , whereas a decrease of K m and k were observed. Furthermore, it can be observed from fluorescence and CD spectra that PEF treatment had increased the number of amino acid residues, especially that of tryptophan, on α-amylase surface with enhanced α-helices by 34.76 % and decreased random coil by 12.04 % on α-amylase when compared with that of untreated. These changes in structure had positive effect on enhancing α-amylase activity and property.

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