Kharkova Anna, Kuznetsova Lyubov, Perchikov Roman, Gertsen Maria, Melnikov Pavel, Zaitsev Nikolay, Zhang Jun, Arlyapov Vyacheslav
The Research Center «BioChemTech», Tula State University, 300012 Tula, Russia.
Laboratory of Soil Chemistry and Ecology, Tula State Lev Tolstoy Pedagogical University, 300026 Tula, Russia.
Gels. 2025 May 12;11(5):355. doi: 10.3390/gels11050355.
A biosensor for the determination of glucose, lactate, ethanol and starch in beverages has been developed using enzymes immobilized by a redox-active gel on a screen-printed electrode. A significant improvement proposed for multichannel biosensors, overcoming stability and sensitivity issues by covalently binding phenazine mediators to a biocompatible protein hydrogel, enhancing the packaging of the enzyme. Glucose oxidase (GOx), alcohol oxidase (AOx) and lactate oxidase (LOx) were used as biological materials, as well as a mixture of GOx with γ-amylase (Am). Redox gels were synthesized from bovine serum albumin (BSA) and phenazine derivatives. It was shown that a neutral red-based redox gel combined with single-walled carbon nanotubes is more promising than other substrates for enzyme immobilization. The lower limit of quantification for glucose, ethanol, lactate and starch using these systems is 0.035 mM, 2.3 mM, 15 mM and 2 mg/L, respectively. Biosensors were used to analyze the content of these substances in alcoholic, kvass and fermentation mass. Statistical analysis of the results showed that the values of glucose, ethanol, lactic acid and starch determined using biosensors and obtained by reference methods differ insignificantly. A set of biosensors developed on the basis of specifically selected enzymes is effective for controlling biotechnological processes and can be used as an alternative to classical analytical methods.
一种用于测定饮料中葡萄糖、乳酸、乙醇和淀粉的生物传感器已被开发出来，它是利用氧化还原活性凝胶固定在丝网印刷电极上的酶制成的。针对多通道生物传感器提出了一项重大改进，即通过将吩嗪介质共价结合到生物相容性蛋白质水凝胶上，克服稳定性和灵敏度问题，增强酶的封装。葡萄糖氧化酶（GOx）、醇氧化酶（AOx）和乳酸氧化酶（LOx）被用作生物材料，以及GOx与γ-淀粉酶（Am）的混合物。氧化还原凝胶由牛血清白蛋白（BSA）和吩嗪衍生物合成。结果表明，基于中性红的氧化还原凝胶与单壁碳纳米管结合比其他酶固定化底物更有前景。使用这些系统测定葡萄糖、乙醇、乳酸和淀粉的定量下限分别为0.035 mM、2.3 mM、15 mM和2 mg/L。生物传感器用于分析酒精饮料、克瓦斯和发酵物料中这些物质的含量。结果的统计分析表明，使用生物传感器测定并通过参考方法获得的葡萄糖、乙醇、乳酸和淀粉的值差异不显著。基于特定选择的酶开发的一组生物传感器对于控制生物技术过程是有效的，并且可以用作经典分析方法的替代方法。
