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超声化学对市售脂肪酶活性和构象的影响

Sonochemical Effect on Activity and Conformation of Commercial Lipases.

作者信息

Nadar Shamraja S, Rathod Virendra K

机构信息

Department of Chemical Engineering, Institute of Chemical Technology, Matunga (E), Mumbai, 400019, India.

出版信息

Appl Biochem Biotechnol. 2017 Apr;181(4):1435-1453. doi: 10.1007/s12010-016-2294-2. Epub 2016 Dec 1.

Abstract

The enzyme under lower-intensity ultrasonic irradiation leads to favourable conformational changes, thereby enhancing its activity. The augmentation of activity of ultrasound-treated enzyme is strongly dependent on ultrasound intensity, duty cycle and exposure time, which was investigated for commercial lipases. Thermomyces lanuginosus (TL) lipase showed a 1.3-fold enhanced activity after irradiating at 22 kHz and 11.38 W cm with 50 % duty cycle for 25-min ultrasonic treatment and 1.5-fold enhanced activity was observed for lipozyme (candida antarctica lipase B (CALB)) lipase, at 22 kHz and 15.48 W cm with 66.67 % duty cycle for 20-min ultrasonic treatment. After sonication, thermodynamic parameters viz. E , ΔH, ΔS and ΔG were evaluated and values were found to be significantly lower for both lipases. In addition, the changes in secondary structure due to sonication were investigated by using Fourier transform infrared (FT-IR), which revealed increase in a certain number of random coiled structure, loss of β-sheets, β-turns and α-helix content in TL lipase and CALB lipase. Also, fluorescence spectroscopy exhibited the increased number of tryptophan on surface of both lipases. Moreover, particle size distribution after sonication also helped to improve surface area and enhanced mass transfer, which contributed to improvement in lipase activity.

摘要

在较低强度超声辐照下,酶会发生有利的构象变化,从而提高其活性。超声处理后酶活性的增强强烈依赖于超声强度、占空比和暴露时间,这已针对商业脂肪酶进行了研究。嗜热栖热菌(TL)脂肪酶在22kHz、11.38W/cm²、50%占空比下进行25分钟超声处理后,活性增强了1.3倍;而Lipozyme(南极假丝酵母脂肪酶B(CALB))脂肪酶在22kHz、15.48W/cm²、66.67%占空比下进行20分钟超声处理后,活性增强了1.5倍。超声处理后,对热力学参数即E、ΔH、ΔS和ΔG进行了评估,发现两种脂肪酶的这些值均显著降低。此外,通过傅里叶变换红外光谱(FT-IR)研究了超声处理引起的二级结构变化,结果显示TL脂肪酶和CALB脂肪酶中一定数量的无规卷曲结构增加,β-折叠、β-转角和α-螺旋含量减少。而且,荧光光谱显示两种脂肪酶表面色氨酸数量增加。此外,超声处理后的粒度分布也有助于增加表面积并增强传质,这有助于脂肪酶活性的提高。

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