Electrochemical Measurement of Immobilized Onion Leaves PPO in Agar-Abelmoschus Escucentus Gum
Onion leaves monophenol monooxygenase and o-diphenol oxidase were extracted by salt and solvent method followed by purification on affinity column using natural affiant. For the fabrication of enzyme-based biosensor, composite matrix of natural biopolymers agar-Abelmoschus escucentus gum was used to...
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Published in: | Procedia chemistry 2012, Vol.6, p.27-35 |
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Main Author: | |
Format: | Article |
Language: | English |
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Online Access: | Get full text |
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Summary: | Onion leaves monophenol monooxygenase and o-diphenol oxidase were extracted by salt and solvent method followed by purification on affinity column using natural affiant. For the fabrication of enzyme-based biosensor, composite matrix of natural biopolymers agar-Abelmoschus escucentus gum was used to immobilize monophenol monooxygenase and o-diphenol oxidase. L-tyrosine and L-dopa were used as substrate for the measurement of phenol. Monophenol monooxygenase and o-diphenol oxidase oxidizes phenolic substrate to the corresponding quinone to allow convenient low-potential detection of phenolic analyte. In the enzymatic catalysis, reduction of quinone to phenol was also observed using (vs.Ag/AgCl) at a platinum electrode. The Vmax for tyrosine and dopa is 102.4 and 108.2μM/litre/min where as Km is 11.26 X 10-3mM and 11.90 X 10-3 mM respectively. The biosensor exhibited good performance in terms of reusability, linearity, sensitivity, fabrication, simplicity, shelf-life and operational stability. Thus, the biosensor is suitable for the analytical quantification of phenols. |
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ISSN: | 1876-6196 1876-6196 |
DOI: | 10.1016/j.proche.2012.10.127 |