Institute of Food Science and Technology, National Taiwan University, Taipei, Taiwan, ROC.
Biosens Bioelectron. 2010 Sep 15;26(1):118-25. doi: 10.1016/j.bios.2010.05.022. Epub 2010 May 21.
β(1→3)-d-Glucan sensors were fabricated using bi-enzyme and tri-enzyme immobilized systems with gold nanoparticles (GNPs) to increase sensitivity. The plant β(1→3)-D-glucanase (βG), glucose oxidase (GOD) or/and peroxidase (POD) in agarose-corn flour-gelatin (ACG) matrix were coated on platinum disc electrode to detect soluble β(1→3)-D-glucan. The atomic force microscopy (AFM) revealed that GNPs embedded in ACG formed tiny islands/clusters with enzymes. Both of bi-enzyme sensor (ACG-βG-GOD-GNPs/Pt) and tri-enzyme sensor (ACG-βG-GOD-POD-GNPs/Pt) had response time less than 20s for β(1→3)-D-glucan. A linear calibration plot for bi-enzyme sensor was obtained for β(1→3)-D-glucan concentration ranged from 100 to 1000 ngmL(-1) (R(2)=0.983). The lower detection limit was 30 ngmL(-1) using applied potential of 200 mV and scan rate of 50 mVs(-1); with signal to noise ratio (S/N) of 3. Fabricated tri-enzyme sensor was also operable under similar conditions with LOD of 50 ngmL(-1) (r(2)=0.989) at -175 mV applied potential and scan rate of 50 mVs(-1). Both sensors were durable and could be repeatedly used for at least 14 times. When the tri-enzyme sensor was employed to analyze β(1→3)-d-glucan content in alcoholic beverages, the results were comparable to those obtained by standard method.
β(1→3)-D-葡聚糖传感器采用双酶和三酶固定化系统与金纳米粒子 (GNPs) 制备，以提高灵敏度。将植物β(1→3)-D-葡聚糖酶 (βG)、葡萄糖氧化酶 (GOD) 或/和过氧化物酶 (POD) 包埋在琼脂-玉米粉-明胶 (ACG) 基质中，涂覆在铂盘电极上，以检测可溶性β(1→3)-D-葡聚糖。原子力显微镜 (AFM) 显示，GNPs 嵌入 ACG 中形成带有酶的微小岛/簇。双酶传感器 (ACG-βG-GOD-GNPs/Pt) 和三酶传感器 (ACG-βG-GOD-POD-GNPs/Pt) 对β(1→3)-D-葡聚糖的响应时间均小于 20s。对于双酶传感器，β(1→3)-D-葡聚糖浓度在 100 至 1000ngmL(-1) 范围内获得了线性校准曲线 (R(2)=0.983)。在 200 mV 应用电位和 50 mVs(-1) 扫描速率下，检测限为 30 ngmL(-1)，信噪比 (S/N) 为 3。在类似条件下，制备的三酶传感器也可操作，在 -175 mV 应用电位和 50 mVs(-1) 扫描速率下，检测限为 50 ngmL(-1) (r(2)=0.989)。两种传感器均耐用，可重复使用至少 14 次。当三酶传感器用于分析酒精饮料中的β(1→3)-d-葡聚糖含量时，结果与标准方法相当。
