Laboratoire de Génie des Procédés Plasma et Traitements de Surface, Université Pierre et Marie Curie-Chimie ParisTech, Paris, France.
Mater Sci Eng C Mater Biol Appl. 2013 Aug 1;33(6):3197-205. doi: 10.1016/j.msec.2013.03.052. Epub 2013 Apr 6.
In this work, a simple and rapid method was used to functionalize carbon electrode in order to efficiently immobilize laccase for biosensor application. A stable allylamine coating was deposited using a low pressure inductively excited RF tubular plasma reactor under mild plasma conditions (low plasma power (10 W), few minutes) to generate high density amine groups (N/C ratio up to 0.18) on rough carbon surface electrodes. The longer was the allylamine plasma deposition time; the better was the surface coverage. Laccase from Trametes versicolor was physisorbed and covalently bound to these allylamine modified carbon surfaces. The laccase activities and current outputs measured in the presence of 2,2'-azinobis-(3-ethylbenzothiazole-6-sulfonic acid) (ABTS) showed that the best efficiency was obtained for electrode plasma coated during 30 min. They showed also that for all the tested electrodes, the activities and current outputs of the covalently immobilized laccases were twice higher than the physically adsorbed ones. The sensitivity of these biocompatible bioelectrodes was evaluated by measuring their catalytic efficiency for oxygen reduction in the presence of ABTS as non-phenolic redox substrate and 2,6-dimethoxyphenol (DMP) as phenolic one. Sensitivities of around 4.8 μA mg(-1)L and 2.7 μA mg(-1)L were attained for ABTS and DMP respectively. An excellent stability of this laccase biosensor was observed for over 6 months.
在这项工作中，采用了一种简单快速的方法对碳电极进行功能化，以便有效地固定漆酶用于生物传感器应用。在温和的等离子体条件下（低等离子体功率（10 W），几分钟），使用低压感应激发射频管状等离子体反应器沉积稳定的烯丙胺涂层，以在粗糙的碳表面电极上生成高密度的胺基团（N/C 比高达 0.18）。烯丙胺等离子体沉积时间越长，表面覆盖率越好。来自变色栓菌的漆酶通过物理吸附和共价结合到这些经烯丙胺修饰的碳表面上。在存在 2,2'-联氮双（3-乙基苯并噻唑-6-磺酸）（ABTS）的情况下测量的漆酶活性和电流输出表明，在 30 分钟内对电极进行等离子体涂覆时获得了最佳效率。它们还表明，对于所有测试的电极，共价固定化漆酶的活性和电流输出都比物理吸附的漆酶高两倍。通过测量在 ABTS 作为非酚类氧化还原底物和 2,6-二甲氧基苯酚（DMP）作为酚类底物存在下对氧还原的催化效率来评估这些生物相容的生物电极的灵敏度。对于 ABTS 和 DMP，分别获得了约 4.8 μA mg(-1)L 和 2.7 μA mg(-1)L 的灵敏度。这种漆酶生物传感器具有出色的稳定性，可观察到超过 6 个月。
