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Chemically amplified kojic acid responses of tyrosinase-based biosensor, based on inhibitory effect to substrate recycling driven by tyrosinase and -ascorbic acid
Authors:Yasushi Hasebe   Katsuhiro Oshima   Osamu Takise  Shunichi Uchiyama
Abstract:A tyrosinase-based chemically amplified biosensor, based on the substrate recycling of polyphenols driven by tyrosinase-catalyzed oxidation and chemical reduction by -ascorbic acid (AsA), has been utilized for the highly sensitive detection of inhibitors of tyrosinase such as kojic acid, benzoic and SCN ion. The amplified current response of immobilized tyrosinase-coupling oxygen electrode due to the recycling was suppressed by the addition of inhibitors, and a highly amplified response to kojic acid over other inhibitors was observed in the presence of 5 mM AsA. The amplification factor (AF) of kojic acid is substantially proportional to the AF of substrates, and the AF for 1 × 10−7 M kojic acid was increased by up to a factor of 143 when 1 × 10−5 M dopamine was used as a competitive substrate in the presence of 5 mM AsA. The amplified calibration curve of kojic acid obtained with 5 mM AsA was shifted towards more than a two decades lower concentration range compared with that of the non-amplified response, and the detection limit of kojic acid was lowered to 7 × 10−8 M.
Keywords:Biosensors   Tyrosinase   Substrate recycling   -Ascorbic acid
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