• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.413-419
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• 1998

Aryl acylamidase [EC 3.5.1.13] present in an acetaminophen-assimilating Pseudomonas sp. has been purified to a homogeneity using series of ammonium sulfate fractionation, DEAE-Sephacel anion exchange, Phenyl-Sepharose CL-4B hydrophobic, and Sephadex G-100 gel-permeation chromatography. The molecular weight, which was estimated by gel-permeation filtration and sodium dodecyl sulfate polyacylamide gel electrophoresis, was about 57 kDa and 56 kDa, respectively, indicating that this enzyme is a monomeric protein. The optimum pH was 10.5 and the optimum temperature was 40$^{\circ}C$. After incubation of the enzyme at 50$^{\circ}C$ for 30 min, residual activity of the enzyme was 34% compared to its original activity. The Km values for acetaminophen and 4'-nitroactanilide were 0.10 mM and 0.11 mM, respectively.

• Journal of Sensor Science and Technology
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• v.17 no.1
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• pp.29-34
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• 2008

A new disposable amperometric tri-enzyme biosensor for the detection of paracetamol has been developed. The paracetamol sensors developed uses horseradish peroxidase modified screen-printed carbon electrodes (HRP-SPCEs) coupled with immobilized enzymes, tyrosinase and aryl acylamidase, prepared using a poly (vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ) matrix. Optimization of the experimental parameters has been performed and the paracetamol biosensor showed detection limit for paracetamol is as low as $100{\mu}M$ and the sensitivity of the sensor is $1.46nA{\mu}M^{-1}cm^{-2}$.