Abstract

The use of agrochemicals has increased considerably in recent years, and consequently, there has been increased exposure of ecosystems and human populations to these highly toxic compounds. The study and development of methodologies to detect these substances with greater sensitivity has become extremely relevant. This article describes, for the first time, the use of atomic force spectroscopy (AFS) in the detection of enzyme-inhibiting herbicides. A nanobiosensor based on an atomic force microscopy (AFM) tip functionalised with the acetolactate synthase (ALS) enzyme was developed and characterised. The herbicide metsulfuron-methyl, an ALS inhibitor, was successfully detected through the acquisition of force curves using this biosensor. The adhesion force values were considerably higher when the biosensor was used. An increase of ∼250% was achieved relative to the adhesion force using an unfunctionalised AFM tip. This considerable increase was the result of a specific interaction between the enzyme and the herbicide, which was primarily responsible for the efficiency of the nanobiosensor. These results indicate that this methodology is promising for the detection of herbicides, pesticides, and other environmental contaminants.

Highlights

  • The use of pesticides and fertilisers has increased considerably in recent years due to the elevated global demand for food

  • The use of chemically modified cantilevers has proven to be a promising alternative for the detection of enzyme-inhibiting herbicides, allowing the use of atomic force microscopy (AFM) as a qualitative detection approach

  • By combining tips functionalised with acetolactate synthase (ALS) and force curve measurements, it was possible to detect, for the first time, the herbicide metsulfuron-methyl

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Summary

Introduction

The use of pesticides and fertilisers has increased considerably in recent years due to the elevated global demand for food. When AFS is used for agrochemical analysis, an alternative method for conferring selectivity to a sensor is based on the mechanism of action of the pesticide on the target plant because each pesticide has a unique mechanism of action that usually involves specific binding to a biomolecule. This relevant alternative has been explored by our research group. We studied the detection of the herbicide metsulfuron-methyl, one of the most widely used herbicides in Brazil for post-emergence weed control in wheat fields [48], by employing a new methodology based on the use of AFM tips modified with the enzyme ALS

Expression of Recombinant ALS
Acetolactate Synthase Activity and Herbicide Inhibition Assays
Chemical Functionalisation of Tips and Substrates
Fourier Transform Infrared Spectroscopy
Results and Discussion
Evaluation of the Interaction between the Enzyme and the Herbicide
Conclusions
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