Analytical Biochemistry 2014-07-15

Detection of glycoalkaloids using disposable biosensors based on genetically modified enzymes.

Michelle Arredondo Espinoza, Georges Istamboulie, Ana Chira, Thierry Noguer, Margarita Stoytcheva, Jean-Louis Marty

文献索引：Anal. Biochem. 457 , 85-90, (2014)

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摘要

In this work we present a rapid, selective, and highly sensitive detection of α-solanine and α-chaconine using cholinesterase-based sensors. The high sensitivity of the devices is brought by the use of a genetically modified acetylcholinesterase (AChE), combined with a one-step detection method based on the measurement of inhibition slope. The selectivity was obtained by using butyrylcholinesterase (BChE), an enzyme able to detect these two toxins with differential inhibition kinetics. The enzymes were immobilized via entrapment in PVA-AWP polymer directly on the working electrode surface. The analysis of the resulting inhibition slope was performed employing linear regression function included in Matlab. The high toxicity of α-chaconine compared to α-solanine due to a better affinity to the active site was proved. The inhibition of glycoalkaloids (GAs) mixture was performed over AChE enzyme wild-type AChE and BChE biosensors resulting in the detection of synergism effect. The developed method allows the detection of (GAs) at 50 ppb in potato matrix.Copyright © 2014 Elsevier Inc. All rights reserved.