Enzyme immobilization on glass fiber membrane for detection of halogenated compounds
Enzyme immobilization on glass fiber membrane for detection of halogenated compounds
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酶固定在玻璃纤维膜上用于检测卤代化合物
DOI：
10.1016/j.ab.2020.113971
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发表时间：
2020-11-15
影响因子：
2.9
通讯作者：
Tang, Lixia
中科院分区：
文献类型：
--
作者：
Gul, Ijaz;Wang, Qian;Tang, Lixia
利用无机膜固定化酶已经引起了越来越多的关注，因为它们不仅提高了酶的稳定性，而且还提供了用户友好的生物设备，可以根据不同的应用进行定制。为此，我们探索了玻璃纤维膜固定化酶的适宜性及其在卤代烃检测中的应用。为此，将卤代丙烷脱卤酶(HHEC)和牛血清白蛋白(BSA)交联并固定在玻璃纤维膜上，而不进行膜功能化。固定化HHEC在4℃(固定化67%对游离态8.1%)和30℃(固定化77%对游离态57%)的60天内表现出比游离HHEC更高的储存稳定性。同样，固定化HHEC的耐热性显著提高。以1，3-二氯-2-丙醇(1，3-DCP)和2，3-二溴-1-丙醇(2，3-DBP)为模型分析物，验证了膜固定化酶的实用性。在优化条件下，1，3-DCP和2，3-DBP的检出限分别为0.06 mm和0.09 mm。对河水和湖水样品的加标回收率令人满意，这表明固定化HHEC在水样污染物筛选中的应用潜力。我们的结果表明，所提出的经济和简便的方法可能有助于酶的稳定，并减轻卤代烃污染。
Enzyme immobilization using inorganic membranes has enticed increased attention as they not only improve enzyme stability, but also furnish user-friendly biodevices that can be tailored to different applications. Herein, we explored the suitability of the glass fiber membrane for enzyme immobilization and its application for halocarbon detection. For this, halohydrin dehalogenase (HheC) and bovine serum albumin were crosslinked and immobilized on a glass fiber membrane without membrane functionalization. Immobilized HheC exhibited higher storage stability than its free counterpart over 60 days at 4 degrees C (67% immobilized vs. 8.1% free) and 30 degrees C (77% immobilized vs. 57% free). Similarly, the thermal endurance of the immobilized HheC was significantly improved. The practical utility of the membrane-immobilized enzyme was demonstrated by colorimetric detection of 1,3-dichlom-2-propanol (1,3-DCP) and 2,3-dibromo-1-propanol (2,3-DBP) as model analytes. Under optimized conditions, the detection limits of 0.06 mM and 0.09 mM were achieved for 1,3-DCP and 2,3-DBP, respectively. The satisfactory recoveries were observed with spiked river and lake water samples, which demonstrate the application potential of immobilized HheC for screening contaminants in water samples. Our results revealed that the proposed frugal and facile approach could be useful for enzyme stabilization, and mitigation of halocarbon pollution.