Surface Functionalization by Hydrophobin-EPSPS Fusion Protein Allows for the Fast and Simple Detection of Glyphosate

• Main Authors: Julia Döring, David Rettke, Gerhard Rödel, Tilo Pompe, Kai Ostermann
• Format: Article
• Language: English
• Published: MDPI AG 2019-08-01
• Series: Biosensors
• Subjects: glyphosate; malachite green assay; hydrophobin; EPSPS; immobilization
• Online Access: https://www.mdpi.com/2079-6374/9/3/104

Glyphosate, the most widely used pesticide worldwide, is under debate due to its potentially cancerogenic effects and harmful influence on biodiversity and environment. Therefore, the detection of glyphosate in water, food or environmental probes is of high interest. Currently detection of glyphosate usually requires specialized, costly instruments, is labor intensive and time consuming. Here we present a fast and simple method to detect glyphosate in the nanomolar range based on the surface immobilization of glyphosate’s target enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) via fusion to the hydrophobin Ccg2 and determination of enzyme activity with a malachite green assay, which is a common photometric technique to measure inorganic phosphate (Pi). The assay demonstrates a new approach for a fast and simple detection of pesticides.