Abstract
Abnormal concentrations of amino acids in blood and urine can be indicative of several diseases, including cancer and diabetes. Therefore, analyses that examine amino acid concentrations are useful for the diagnosis of such diseases. In this study, we developed an enzyme-immobilized, small reactor column for flow analysis of amino acid concentrations. For the recognition of asparagine and lysine, asparaginyl-tRNA synthetase and lysyl-tRNA synthase were immobilized onto microparticles, respectively, and coupled with coloration reagents for spectrophotometric detection. This assay has some advantages in the analytical field, such as the ability to detect small amounts of analyte, allowing for the use of a small reaction volume, and ensuring a rapid and efficient reaction rate. This approach provided selective quantitation of up to 480 μM of asparagine and lysine in 200 mM Tris–HCl buffer (pH 8.0).
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Acknowledgments
This work was partly supported by Hiroshima City University Grant for Special Academic Research (General Studies) and also supported by Grant-in-Aid for Scientific Research C (No. 25330344).
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Kugimiya, A., Konishi, H. & Fukada, R. Flow Analysis of Amino Acids by Using a Newly Developed Aminoacyl-tRNA Synthetase–Immobilized, Small Reactor Column–Based Assay. Appl Biochem Biotechnol 178, 924–931 (2016). https://doi.org/10.1007/s12010-015-1918-2
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DOI: https://doi.org/10.1007/s12010-015-1918-2