Enzyme-based electrochemical biosensor for antimonite detection in water

Abstract

Antimonite (Sb^III) is a naturally occurring contaminant demanding on-site ultrasensitive detection. The enzyme-based electrochemical (EC) biosensors are promising, but the lack of specific Sb^III oxidizing enzymes hindered the past efforts. Herein, we modulated the specificity of arsenite oxidase AioAB toward Sb^III by regulating its spatial conformation from tight to loose using the metal-organic framework ZIF-8. The constructed EC biosensor, AioAB@ZIF-8, exhibited the substrate specificity toward Sb^III at 12.8 s^-1 μM^-1, an order of magnitude higher than that of As^III (1.1 s^-1 μM^-1). Relaxing AioAB structure in ZIF-8 was evidenced by the break of the S-S bond and the conversion of α helix to the random coil as suggested by Raman spectroscopy. Our AioAB@ZIF-8 EC sensor exhibited a dynamic linear range in 0.041-4.1 μM at a response time of 5 s, and the detection limit at 0.041 μM at a high sensitivity of 1894 nA μM^-1. The insights into tuning the specificity of an enzyme shed new light on biosensing metal(loid)s without specific proteins.

Keywords: Antimonite; Electrochemical biosensor; Enzyme; Metal-organic frameworks; Substrate specificity.