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Covalent attachment of laccase to carboxymethyl-botryosphaeran in aqueous solution for the construction of a voltammetric biosensor to quantify quercetin.
Bioelectrochemistry ( IF 5 ) Pub Date : 2020-05-15 , DOI: 10.1016/j.bioelechem.2020.107543
Adriana Gomes 1 , Gabriel J Mattos 1 , Bruna Coldibeli 1 , Robert F H Dekker 2 , Aneli M Barbosa Dekker 1 , Elen R Sartori 1
Affiliation  

Laccase from Botryosphaeria rhodina MAMB-05 was covalently immobilized on carboxymethyl-botryosphaeran by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide (EDC/NHS) in aqueous solution. This approach was employed to fabricate a novel laccase-based biosensor to electrochemically quantify quercetin (QCT), using a simple carbon black paste electrode as a transducer. The proposed biosensor was characterized by electrochemical impedance spectroscopy and Nyquist plots were used to evaluate the immobilization of the enzyme. For determining QCT, variables were optimized, that included experimental conditions for laccase immobilization, pH of the supporting electrolyte, and instrumental parameters of the electroanalytical technique. From square-wave-voltammograms, a linear dependence between the cathodic current peak and QCT concentration was observed within the range 4.98–50.0 × 10−8 mol L−1, with a theoretical detection limit of 2.6 × 10−8 mol L−1. The proposed method was successfully applied to determine QCT in beverages, pharmaceuticals, and biological samples. The proposed biosensor device presented good selectivity in the presence of uric acid, various inorganic ions, as well as other phenolic compounds, demonstrating the potential application of this biosensing platform in chemically complex solutions. Operational and analytical stability of the laccase-biosensor were evaluated, and good intra-day (SD = 1.23%) and inter-day (SD = 2.32%) repeatability, and long storage stability (SD = 3.47%) are presented.



中文翻译:

漆酶与水溶液中羧甲基-葡聚糖的共价结合,用于构建伏安生物传感器以定量槲皮素。

葡萄球菌的漆酶MAMB-05通过1-乙基-3-(3-二甲基氨基丙基)碳二亚胺和N-羟基琥珀酰亚胺(EDC / NHS)在水溶液中共价固定在羧甲基-葡聚糖上。使用一种简单的炭黑糊状电极作为换能器,采用该方法来制造新颖的基于漆酶的生物传感器,以电化学定量槲皮素(QCT)。拟议的生物传感器的特征是通过电化学阻抗谱和奈奎斯特图被用来评估酶的固定化。为了确定QCT,对变量进行了优化,包括漆酶固定化的实验条件,支持电解质的pH值以及电分析技术的仪器参数。从方波伏安图-8 mol L -1,理论检测极限为2.6×10 -8 mol L -1。该方法成功应用于饮料,药品和生物样品中的QCT测定。所提出的生物传感器设备在尿酸,各种无机离子以及其他酚类化合物的存在下表现出良好的选择性,证明了该生物传感平台在化学复杂溶液中的潜在应用。评估了漆酶生物传感器的操作和分析稳定性,并显示了良好的日内(SD = 1.23%)和日间(SD = 2.32%)重复性,以及长期保存稳定性(SD = 3.47%)。

更新日期:2020-05-15
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