Choline biosensor constructed with chitinous membrane from soldier crab and its application in measuring cholinesterase inhibitory activities.
An amperometric flow-injection choline biosensor was assembled utilizing natural chitinous membrane as the supporting material for biocatalyst immobilization, and the membrane was purified from Taiwanese soldier crab, Mictyris brevidactylus. The chitinous membrane (<50.0 microm in thickness) was covalently immobilized with choline oxidase (EC 3.1.1.17 from Alcaligenes sp.) and then attached onto the platinum electrode of an amperometric flow cell. The flow cell served as the choline sensing device of the proposed FIA system. The sensor signal (peak height of the FIAgram) was linearly related to choline concentration (r=0.999 for choline up to 5.0mM) with low detection limit (S/N>3 for 10.0 microM choline) and high reproducibility (CV<3% for 1.0mM choline, n=7). The system was proved to be useful in measuring cholinesterase inhibitory activities of synthetic chemicals or natural products.
Hsieh
Bo-Chuan
BC
Department of Bio-Industrial Mechatronics Engineering, National Taiwan University, 136 Chou-Shan Road, Taipei City 106, Taiwan, ROC.
Matsumoto
Kiyoshi
K
Cheng
Tzong-Jih
TJ
Yuu
Gyokurin
G
Chen
Richie L C
RL
D000421:Alcaligenes / Q000201:enzymology; D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D003386:Brachyura; D002686:Chitin / Q000737:chemistry*; D002794:Choline / Q000032:analysis; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004563:Electrochemistry; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000378:metabolism*; D017022:Flow Injection Analysis; D006863:Hydrogen-Ion Concentration; D010936:Plant Extracts / Q000494:pharmacology*; D010984:Platinum; D012680:Sensitivity and Specificity
D002800:Cholinesterase Inhibitors; D004800:Enzymes, Immobilized; D010936:Plant Extracts; D002686:Chitin; D010984:Platinum; D000429:Alcohol Oxidoreductases; C019888:choline oxidase; D002794:Choline