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Nanomaterial-based Novel Methods for the Assay of Enzyme Activity

Author: CaoYa
Tutor: LiGenXi
School: Nanjing University
Course: Biochemistry and Molecular Biology
Keywords: Purine nucleoside phosphorylase Indoleamine 2,3 - dioxygenase Silver nanoparticles Platinum nanoparticles Biocatalysis Electrocatalysis
CLC: TB383.1
Type: Master's thesis
Year: 2011
Downloads: 56
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Abstract


Various chemical reactions in the metabolic processes are almost always conducted in the enzyme-catalyzed activity exception will seriously disrupt the normal activities of life, and is closely related to many diseases, therefore, the detection of activity has attracted widespread attention, and has developed a variety of biosensors. Nanomaterials because of their unique optical, electrical, magnetic properties, and good biocompatibility, can significantly improve the sensitivity of the biosensor, has been widely applied in the field of bio-sensors. The papers silver nanoparticles and platinum nanoparticles were used to construct the detection of purine nucleoside phosphorylase and indoleamine 2,3 - dioxygenase biosensing system. We build two sensors having high sensitivity and high selectivity characteristics, compared to the traditional activity detection method, and provides some technical support for the clinical examination of these two enzymes. 1 based on the the silver nanoparticles purine nucleoside phosphorylase activity detection purine nucleoside phosphorylase is widespread related enzymes involved in purine metabolism in vivo, to play an important role in the nucleic salvage pathway. Chapter OF catalyzed by purine nucleoside phosphorylase guanosine generate guanine enzyme reaction characteristics, combined with the optical and electrochemical properties of the silver nanoparticles, was constructed that contains the optical and electrochemical detection system biosensor of novel enzyme activity. The experimental results show that the sensor has a high sensitivity for the detection of the purine nucleoside phosphorylase activity, a good stability and specificity, can fully meet the needs of the clinical detection. Meanwhile, the detection system to avoid the use of the of other coupling enzyme or isotope-labeled, greatly reduces the complexity of the detection operation, the kinetics of novel inhibitors of purine nucleoside phosphorylase development. 2 based on platinum nanoparticles indoleamine 2,3 - dioxygenase activity detection indoleamine 2,3 - dioxygenase mammalian cytoplasmic catalytic tryptophan along the kynurenine pathway metabolism speed limit enzyme, its active center contains a heme molecule, can catalyze the L-tryptophan in the cutting of the pyrrole ring, and the cutting is inserted at the two oxygen atoms, to thereby form N-formyl-dog urine histidine. Chapter OF biocatalytic process using the enzyme-mediated and platinum nanocolloid particle mediated electrical coupling of catalytic processes, to construct a novel electrochemical sensor for the detection of indoleamine 2,3 - dioxygenase enzyme activity . The experimental results show that only in indoleamine 2,3 - dioxygenase activity exist under the conditions, in order to connect platinum nanoparticles to the electrode surface to obtain its electrical catalyzed hydrogen peroxide and restore the electrochemical signal. The detection system has a higher sensitivity detection limit reached 6.24U/mL. Further studies showed that the system can be successfully applied to the activity detection in complex samples such as serum activity inhibitor effect.

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