کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1173151 | 961654 | 2012 | 5 صفحه PDF | دانلود رایگان |
An online immobilized glucose oxidase (GOx) capillary microreactor was developed based on an enzymatic redox reaction with 1,4-benzoquinone as an acceptor of electrons, replacing the molecular oxygen typically used in a GOx reaction to achieve direct ultraviolet detection without derivation. A high efficiency of enzymolysis was obtained at 1 mg ml−1 1,4-benzoquinone for 5 min of incubation at 25 °C, and baseline separation of the substrate and product could be achieved with a resolution of 3.85 by employing 20 mM phosphate buffer (pH 8.0) containing 40 mg ml−1 sulfated β-cyclodextrin as an additive, a constant voltage of 15 kV, and a detection wavelength of 220 nm. In addition, an online enzyme inhibition study was performed on the immobilized GOx microreactor with metal ions Ag+ and Cu2+ used as model inhibitors. The results indicate that Ag+ (IC50 = 69.16 μM) has a markedly higher inhibitory effect than Cu2+ (IC50 = 1.33 mM). The protocol described can be applied in high-throughput screening of enzyme reactions and inhibitors.
Journal: Analytical Biochemistry - Volume 427, Issue 2, 15 August 2012, Pages 139–143