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首页> 外文期刊>The protein journal >Effects of Pulsed Electric Field (PEF) Treatment on Enhancing Activity and Conformation of alpha-Amylase
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Effects of Pulsed Electric Field (PEF) Treatment on Enhancing Activity and Conformation of alpha-Amylase

机译：脉冲电场（PEF）处理对增强活性和α-淀粉酶构象的影响

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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 alpha-amylase were investigated. Conformational transitions were also studied with the aid of circular dichroism (CD) and fluorescence spectra. The maximum enzymatic activity of alpha-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 +/- A 1.14 % compared with control. The activation effect could last for 18 h at 4 A degrees C. PEF treatment could widen the range of optimum temperature for alpha-amylase, however, it barely exerted any effect on the optimum pH. On the other hand, alpha-amylase treated by PEF showed an increase of V (max), t(1/2) and Delta 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 alpha-amylase surface with enhanced alpha-helices by 34.76 % and decreased random coil by 12.04 % on alpha-amylase when compared with that of untreated. These changes in structure had positive effect on enhancing alpha-amylase activity and property.
机译：为了探索高效，安全，快速地应用脉冲电场（PEF）技术进行酶促修饰，研究了PEF处理对α-淀粉酶的酶促活性，性质和动力学参数的影响。还借助圆二色性（CD）和荧光光谱研究了构象转变。在15 kV / cm电场强度和100 mL / min流速PEF处理下，α-淀粉酶具有最大的酶活性，与对照组相比，酶活性提高了22.13 +/- A 1.14％。活化作用可以在4 A的温度下持续18小时。PEF处理可以扩大α-淀粉酶的最佳温度范围，但是，它几乎没有对最佳pH产生任何影响。另一方面，用PEF处理的α-淀粉酶显示出V（max），t（1/2）和ΔG增加，而观察到K（m）和k降低。此外，从荧光和CD光谱可以看出，PEF处理增加了具有α-螺旋增强的α-淀粉酶表面上的氨基酸残基，特别是色氨酸的氨基酸残基的数量，增加了34.76％，而使α的随机卷曲减少了12.04％ -淀粉酶与未处理的淀粉酶相比。这些结构变化对增强α-淀粉酶活性和性质具有积极作用。

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