fulltext.study @t Gmail

Putrescine biosensor based on putrescine oxidase from Kocuria rosea

Paper ID Volume ID Publish Year Pages File Format Full-Text
17214 42652 2012 5 PDF Available
Title
Putrescine biosensor based on putrescine oxidase from Kocuria rosea
Abstract

The novel putrescine oxidase based amperometric biosensor selectively measures putrescine, which can be considered as an indicator of microbial spoilage. Putrescine oxidase (PUOX, EC 1.4.3.10) was isolated from Kocuria rosea (Micrococcus rubens) by an improved and simplified purification process. Cells were grown on brain heart infusion medium supplemented with putrescine. Cell-free extract was prepared in Tris buffer (pH 8.0) by Bead-beater. A newly elaborated step based on three-phase partitioning (TPP) was applied in the purification protocol of PUOX.The purified enzyme was immobilized on the surface of a spectroscopic graphite electrode in redox hydrogel with horseradish peroxidase, Os mediator and poly(ethylene glycol) (400) diglycidyl ether (PEGDGE) as crosslinking agent. This modified working electrode was used in wall-jet type amperometric cell together with the Ag/AgCl (0.1 M KCl) reference electrode and a platinum wire as auxiliary electrode in flow injection analysis system (FIA). Hydrogel composition, pH and potential dependence were studied. Optimal working conditions were 0.45 mL min−1 flow rate of phosphate buffer (66 mM, pH 8.0) and +50 mV polarizing potential vs. Ag/AgCl. The linear measuring range of the method was 0.01–0.25 mM putrescine, while the detection limit was 5 μM. Beer samples were investigated by the putrescine biosensor and the results were compared by those of HPLC reference method.

► New enzyme purification procedure based on three-phase partitioning. ► Novel biosensor for selective determination of putrescine for food quality control. ► Novel modified electrode based on putrescine oxidase, peroxidase and Os complex. ► Optimized operation of unique enzyme combination. ► The putrescine biosensor was successfully used for the analysis of beer samples.

Keywords
Biosensor; Putrescine; Putrescine oxidase isolation; Kocuria rosea; Three-phase partitioning; Electrochemical sensor
First Page Preview
Putrescine biosensor based on putrescine oxidase from Kocuria rosea
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 51, Issue 5, 10 October 2012, Pages 258–262
Authors
, , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us