Acetylcholinesterase with mesoporous silica: Covalent immobilization, physiochemical characterization, and its application in food for pesticide detection

J Cell Biochem. 2019 Jun;120(6):10777-10786. doi: 10.1002/jcb.28369. Epub 2019 Jan 23.

Abstract

Toxic contamination of commonly consumed food products and water due to food chain vulnerability via agricultural products and commodities is a serious health hazard. This study reports on Santa Barbara Amorphous (SBA-15), a type of mesoporous silica nanoparticles, for efficient and stable acetylcholinesterase (AChE) adhesion toward detection of toxic pesticides. AChE was immobilized to the inert framework of mesoporous materials viz. SBA-15 with a proficient hydrolytic response toward acetylthiocholine. The immobilized system acts as a biosensor for the detection of pesticides, which are organophosphorus compounds in food. Both the SBA-15 and immobilized SBA-15 were characterized to give an insight on the physiochemical and morphological modification properties. The enzyme activity was accessed by Ellman's spectrophotometric bioassay for bare and enzyme-immobilized SBA-15 that resulted in promising enzymatic activity with the counterpart. Enzyme stability was also studied, which exhibited that immobilized AChE retained its catalytic activity up to 60 days and retained 80% of the hydrolytic activity even at 37°C. On the basis of the success of immobilized enzyme (covalent) being inhibited by acetylthiocholine, the sensor was administered for the inhibition by monocrotophos and dimethoate that are used widely as pesticides in agricultural. The inhibitory concentration (IC50 ) value was found to be 2.5 ppb for monocrotophos and 1.5 ppb for dimethoate inhibiting immobilized AChE. This was verified using cyclic voltammetry, an electrochemical analysis thus proving that the SBA-15@AChE complex could be used as a sensitive and highly stable sensor for detecting the concentration of hazardous pesticide compounds.

Keywords: Santa Barbara Amorphous; acetylcholinesterase; acetylthiocholine; covalent immobilization; enzyme stability; inhibitory concentration 50; pesticides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholinesterase / chemistry*
  • Acetylthiocholine / chemistry
  • Adsorption
  • Biosensing Techniques / methods
  • Carbonated Beverages / analysis
  • Dimethoate / analysis*
  • Electrochemical Techniques*
  • Enzyme Assays
  • Enzymes, Immobilized / chemistry*
  • Food Contamination / analysis
  • Humans
  • Monocrotophos / analysis*
  • Nanoparticles / chemistry
  • Pesticides / analysis*
  • Porosity
  • Sensitivity and Specificity
  • Silicon Dioxide / chemistry

Substances

  • Enzymes, Immobilized
  • Pesticides
  • SBA-15
  • Acetylthiocholine
  • Monocrotophos
  • Silicon Dioxide
  • Acetylcholinesterase
  • Dimethoate