Department of Biochemistry, Faculty of Science, Ege University, 35100 Bornova-Izmir, Turkiye.
Biotechnol Prog. 2011 Mar-Apr;27(2):530-8. doi: 10.1002/btpr.544. Epub 2011 Jan 10.
This article deals with the use of pyranose oxidase (PyOx) and glucose oxidase (GOx) enzymes in amperometric biosensor design and their application in monitoring fermentation processes with the combination of flow injection analysis (FIA). The amperometric studies were carried out at -0.7 V by following the oxygen consumption due to the enzymatic reactions for both batch and FIA modes. Optimization studies (enzyme amounts and pH) and analytical parameters such as linearity, repeatability, effect of interference, storage, and operational stabilities have been studied. Under optimized conditions, for the PyOx-based biosensor, linear graph was obtained from 0.025 to 0.5 mM glucose in phosphate buffer (50 mM) at pH 7.0 with the equation of y = 3.358x + 0.028 and R(2) = 0.998. Linearity was found to be 0.01-1.0 mM in citrate buffer (50 mM and pH 4.0) with the equation of y = 1.539x + 0.181 and R(2) = 0.992 for the GOx biosensor. Finally, these biosensor configurations were further evaluated in a conventional flow injection system. Results from batch experiments provide a guide to design sensitive, stable, and interference-free biosensors for FIA mode. Biosensor stability, dynamic range, and repeatability were also studied in FIA conditions, and the applicability for the determination of glucose in fermentation medium could be successfully demonstrated. The FIA-combined glucose biosensor was used for the offline monitoring of yeast fermentation. The obtained results correlated well with HPLC measurements.
本文探讨了吡喃糖氧化酶(PyOx)和葡萄糖氧化酶(GOx)在安培生物传感器设计中的应用,以及它们与流动注射分析(FIA)相结合在监测发酵过程中的应用。在 -0.7 V 的电势下进行安培研究,通过跟踪酶反应引起的氧气消耗来监测分批和 FIA 两种模式下的反应。对优化研究(酶用量和 pH 值)和分析参数(线性度、重复性、干扰影响、存储和操作稳定性)进行了研究。在优化条件下,基于 PyOx 的生物传感器在磷酸盐缓冲液(50 mM,pH 7.0)中,葡萄糖浓度在 0.025 到 0.5 mM 之间时,得到了线性图,其方程式为 y = 3.358x + 0.028,R² = 0.998。在柠檬酸缓冲液(50 mM,pH 4.0)中,线性范围为 0.01-1.0 mM,其方程式为 y = 1.539x + 0.181,R² = 0.992。最后,在传统的流动注射系统中进一步评估了这些生物传感器的配置。批量实验结果为设计用于 FIA 模式的敏感、稳定且无干扰的生物传感器提供了指导。还在 FIA 条件下研究了生物传感器的稳定性、动态范围和重复性,并成功证明了其在发酵培养基中葡萄糖测定的适用性。FIA 结合的葡萄糖生物传感器用于酵母发酵的离线监测。获得的结果与 HPLC 测量结果相关性良好。