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Rapid enzymatic assays for l-citrulline and l-arginine based on the platform of pyrophosphate detection

Paper ID Volume ID Publish Year Pages File Format Full-Text
17059 42637 2014 6 PDF Available
Title
Rapid enzymatic assays for l-citrulline and l-arginine based on the platform of pyrophosphate detection
Abstract

•Rapid enzymatic assays for l-citrulline and l-arginine were developed.•Higher selectivity than that of conventional enzymatic and colorimetric assays.•Accurate quantification even in the presence of a biological sample.•Usefulness of the pyrophosphate detection system as a platform of selective assays.

Rapid determination of l-citrulline and l-arginine, physiologically important amino acids, is a beneficial technique from the scientific and medical viewpoints. In this study, enzymatic assays for l-citrulline and l-arginine were established and evaluated. l-Citrulline assay was constructed by coupling argininosuccinate synthetase to a pyrophosphate detection system, in which pyruvate phosphate dikinase was employed, so that the citrulline-dependent production of pyrophosphate could be determined. Furthermore, the l-arginine assay was developed by coupling arginine deiminase to the l-citrulline assay. Both assays exhibited high selectivity to l-citrulline and l-arginine without any significant reactivity to other proteinaceous amino acids. These assays were also resistant to various contaminants that interfered with the conventional l-citrulline and l-arginine assays. The high accuracy of these assays was demonstrated by measurements in the presence of human plasma. Because these assays can be conducted under the neutral pH without terminating the reaction progress, they allow not only measurements in static analyte solutions, but also real-time monitoring of l-citrulline and l-arginine synthesis in the reaction mixture. The features of these assays also demonstrated that the pyrophosphate detection system served as a useful platform to develop selective and robust enzymatic assays by being coupled to a pyrophosphate-producing enzyme.

Keywords
ADI, arginine deiminase; ASS, argininosuccinate synthetase; BSA, bovine serum albumin; CV, coefficient of variation; GdmCl, guanidinium chloride; HRP, horseradish peroxidase; NOS, nitric oxide synthase; PEP, phosphoenolpyruvate; POX, pyruvate oxidase; PPD
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Rapid enzymatic assays for l-citrulline and l-arginine based on the platform of pyrophosphate detection
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 57, 10 April 2014, Pages 36–41
Authors
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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