Design of acetylcholinesterases for biosensor applications.
In recent years, the use of acetylcholinesterases (AChEs) in biosensor technology has gained enormous attention, in particular with respect to insecticide detection. The principle of biosensors using AChE as a biological recognition element is based on the inhibition of the enzyme's natural catalytic activity by the agent that is to be detected. The advanced understanding of the structure-function-relationship of AChEs serves as the basis for developing enzyme variants, which, compared to the wild type, show an increased inhibition efficiency at low insecticide concentrations and thus a higher sensitivity. This review describes different expression systems that have been used for the production of recombinant AChE. In addition, approaches to purify recombinant AChEs to a degree that is suitable for analytical applications will be elucidated as well as the various attempts that have been undertaken to increase the sensitivity of AChE to specified organophosphates and carbamates using side-directed mutagenesis and employing the enzyme in different assay formats.
Schulze
Holger
H
Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany.
Vorlová
Sandra
S
Villatte
Francois
F
Bachmann
Till T
TT
Schmid
Rolf D
RD
D000110:Acetylcholinesterase / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002800:Cholinesterase Inhibitors / Q000032:analysis* / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004867:Equipment Design; D005786:Gene Expression Regulation; D007306:Insecticides / Q000032:analysis; D009943:Organophosphorus Compounds; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D002800:Cholinesterase Inhibitors; D004800:Enzymes, Immobilized; D007306:Insecticides; D009943:Organophosphorus Compounds; D011994:Recombinant Proteins; D000110:Acetylcholinesterase