Department of Biochemistry, MD University, Rohtak 124 001, India.
Int J Biol Macromol. 2013 Mar;54:250-7. doi: 10.1016/j.ijbiomac.2012.12.004. Epub 2012 Dec 10.
A method is described for construction of a highly sensitive amperometric L-amino acid biosensor based on covalent immobilization of goat kidney L-amino acid oxidase (LAAO) onto carboxylated multiwalled carbon nanotubes/nickel hexacyanoferrate/polypyrrole hybrid film electrodeposited on surface of glassy carbon electrode. The biosensor showed optimum response within 5s at pH 7.5 and 35 °C, when polarized at 0.15 V vs. Ag/AgCl. There was a linear relationship between biosensor response (μA) and L-phenylalanine concentration in the range, 0.5 μM to 100 mM. The sensitivity of the biosensor was 79.31 nA cm(-2) μM(-1) with a detection limit of 0.5 μM (S/N=3). The enzyme electrode was used 160 times over a period of 140 days, when stored dry at 4 °C. The biosensor was evaluated and employed for determination of L-amino acid level in fruit juices and alcoholic beverages. The biosensor showed excellent analytical characteristics along with short response time, good shelf life, and negligible interference of commonly present interferents. The film could be used for improvement of other biosensors also.
描述了一种基于共价固定化羊肾 L-氨基酸氧化酶(LAAO)到羧基化多壁碳纳米管/镍六氰合铁酸盐/聚吡咯杂合膜的高灵敏度安培 L-氨基酸生物传感器的构建方法,该杂合膜沉积在玻碳电极表面。当在 35°C 、pH7.5 条件下,在 0.15V(相对于 Ag/AgCl)下极化时,生物传感器在 5s 内显示出最佳响应。生物传感器响应(μA)与 L-苯丙氨酸浓度在 0.5μM 至 100mM 范围内呈线性关系。生物传感器的灵敏度为 79.31nAcm(-2)μM(-1),检测限为 0.5μM(S/N=3)。当在 4°C 下干燥储存时,酶电极在 140 天内使用了 160 次。该生物传感器用于果汁和酒精饮料中 L-氨基酸水平的测定,具有出色的分析特性,响应时间短,保质期长,并且常见干扰物的干扰可忽略不计。该膜还可用于改进其他生物传感器。