Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya St., 3, 142290 Puschino, Moscow Region, Russia.
Int J Mol Sci. 2023 Apr 5;24(7):6766. doi: 10.3390/ijms24076766.
Phenols are widely used in industries despite their toxicity, which requires governments to limit their concentration in water to 5 mg/L before discharge to the city sewer. Thus, it is essential to develop a rapid, simple, and low-cost detection method for phenol. This study explored two pathways of peroxidase immobilization to develop a phenol detection system: peroxidase encapsulation into polyelectrolyte microcapsules and peroxidase captured by CaCO. The encapsulation of peroxidase decreased enzyme activity by 96%; thus, this method cannot be used for detection systems. The capturing process of peroxidase by CaCO microspherulites did not affect the maximum reaction rate and the Michaelis constant of peroxidase. The native peroxidase-Vmax = 109 µM/min, Km = 994 µM; CaCO-peroxidase-Vmax = 93.5 µM/min, Km = 956 µM. Ultimately, a reusable phenol detection system based on CaCO microparticles with immobilized peroxidase was developed, capable of detecting phenol in the range of 700 ng/mL to 14 µg/mL, with an error not exceeding 5%, and having a relatively low cost and production time. The efficiency of the system was confirmed by determining the content of phenol in a paintwork product.
酚类广泛应用于工业中,尽管它们具有毒性,但这要求政府在将其排放到城市污水管之前,将水中的浓度限制在 5mg/L 以下。因此,开发一种快速、简单且低成本的酚类检测方法至关重要。本研究探索了两种固定化过氧化物酶的途径,以开发酚类检测系统:过氧化物酶包封在聚电解质微胶囊中和过氧化物酶被 CaCO 捕获。过氧化物酶的包封使酶活性降低了 96%;因此,该方法不能用于检测系统。CaCO3 微球粒对过氧化物酶的捕获过程并不影响过氧化物酶的最大反应速率和米氏常数。天然过氧化物酶-Vmax = 109 µM/min,Km = 994 µM;CaCO3-过氧化物酶-Vmax = 93.5 µM/min,Km = 956 µM。最终,开发了一种基于固定化过氧化物酶的 CaCO3 微球粒的可重复使用的酚类检测系统,能够检测 700ng/mL 至 14 µg/mL 范围内的酚类,误差不超过 5%,且具有相对较低的成本和生产时间。通过测定木制品产品中的酚类含量,验证了该系统的效率。