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首页> 外文期刊>Analytical chemistry >Covalent immobilization of beta-galactosidase onto a gold-coated magnetoelastic transducer via a self-assembled monolayer: Toward a magnetoelastic biosensor
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Covalent immobilization of beta-galactosidase onto a gold-coated magnetoelastic transducer via a self-assembled monolayer: Toward a magnetoelastic biosensor

机译：通过自组装单层将β-半乳糖苷酶共价固定在镀金的磁弹性换能器上：朝向磁弹性生物传感器

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摘要

The enzyme beta-galactosidase has been covalently immobilized onto a gold-coated magnetoelastic film via a self-assembled monolayer (SAM) of omega-carboxylic acid alkylthiol. Use of magnetoelastic transduction allows for the wireless monitoring of enzymatic activity through the associated change in the frequency and amplitude of magnetic fields. The formations of SAMs of 3-mercaptopropanoic acid and thioctic acid were monitored by magnetoelastic transduction. After coupling of beta-galactosidase to the SAMs, the enzyme activity was monitored by using a substrate that forms an insoluble product upon action of the enzyme. Specifically, an indolyl galactopyranoside substrate was employed in conjunction with an azo dye as the precipitating system. The immobilized enzyme was evaluated and found to have an apparent Michaelis-Menten constant (K-M) of 1.2 mM for the indolyl galactopyranoside. Calibration plots for both substrates and inhibitors were generated to establish the versatility of this sensing system. Kinetic parameters for nonprecipitating substrates were determined in conjunction with a precipitating enzymatic substrate by way of a competitive inhibition study using beta-galactosidase attached to magnetoelastic strips. The methods developed within this work allow for the fabrication of wireless enzyme sensing systems, which can also be used as another means of screening for enzyme inhibitors. [References: 20]
机译：β-半乳糖苷酶已通过ω-羧酸烷基硫醇的自组装单分子层（SAM）共价固定在涂金的磁弹性膜上。磁弹性转导的使用允许通过磁场频率和振幅的相关变化来无线监测酶活性。通过磁弹性转导监测了3-巯基丙酸和硫辛酸的SAM的形成。在将β-半乳糖苷酶偶联至SAM之后，通过使用在酶作用下形成不溶产物的底物来监测酶活性。具体地，将吲哚基吡喃半乳糖苷底物与偶氮染料一起用作沉淀体系。对固定化的酶进行了评估，发现吲哚基吡喃半乳糖苷的表观Michaelis-Menten常数（K-M）为1.2 mM。生成了底物和抑制剂的标定图，以建立该传感系统的多功能性。通过竞争抑制研究，使用附着在磁弹性条上的β-半乳糖苷酶，结合沉淀的酶促底物确定了非沉淀底物的动力学参数。在这项工作中开发的方法允许制造无线酶传感系统，该系统也可以用作筛选酶抑制剂的另一种方法。 [参考：20]

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