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首页> 外文期刊>Electroanalysis >Pure organic phase phenol biosensor based on tyrosinase entrapped in a vapor deposited titania sol-gel membrane
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Pure organic phase phenol biosensor based on tyrosinase entrapped in a vapor deposited titania sol-gel membrane

机译：气相沉积二氧化钛溶胶-凝胶膜中捕获的基于酪氨酸酶的纯有机相酚生物传感器

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摘要

A novel amperometric biosensor was constructed for the determination of phenols in pure organic phase. This biosensor was fabricated by immobilizing tyrosinase in a titania sol-gel membrane which was obtained with a vapor deposition method. This method was facile and avoided the calcination step needed in conventional titania sol-gel process. The titania sol-gel membrane could effectively retain the essential water layer around the enzyme molecule needed for maintaining its activity in organic phase. The experimental parameters such as solvent and operating potential were optimized. At - 100 mV this biosensor showed a good amperometric response to phenols in pure chloroform without any mediator and rehydration of the enzyme. For catechol determination the sensor exhibited a fast response of less than 5 seconds. The sensitivity of different phenols was as follows: catechol > phenol > p-cresol. Additionally, the apparent Michaelis-Menten constants of the encapsulated tyrosinase to catechol, phenol and p-cresol were found to be 0.15 +/- 0.003, 0.17 +/- 0.008 and 0.21 +/- 0.004 mM, respectively. The biosensor had also good reproducibility and stability. This work provided a promising platform for the construction of pure organic phase biosensors and the determination of substrates with poor water solubility.
机译：构建了一种新型的安培生物传感器，用于测定纯有机相中的苯酚。通过将酪氨酸酶固定在通过气相沉积法获得的二氧化钛溶胶-凝胶膜中来制造该生物传感器。该方法是容易的，并且避免了常规二氧化钛溶胶-凝胶法中所需的煅烧步骤。二氧化钛溶胶-凝胶膜可以有效地在酶分子周围保留必需的水层，以维持其在有机相中的活性。优化了实验参数，例如溶剂和工作电位。在-100 mV时，该生物传感器对纯氯仿中的苯酚显示出良好的安培响应，而没有任何介体和酶的再水化作用。对于邻苯二酚的测定，传感器表现出不到5秒的快速响应。不同苯酚的敏感性如下：儿茶酚>苯酚>对甲酚。另外，发现包封的酪氨酸酶对儿茶酚，苯酚和对甲酚的表观Michaelis-Menten常数分别为0.15 +/- 0.003、0.17 +/- 0.008和0.21 +/- 0.004mM。该生物传感器还具有良好的再现性和稳定性。这项工作为构建纯有机相生物传感器以及确定水溶性差的底物提供了一个有前途的平台。

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