Immobilization of phenylalanine dehydrogenase and its application in flow-injection analysis system for determination of plasma phenylalanine

Applied Biochemistry and Biotechnology
L Tarhan, Hulya Ayar-Kayali

Abstract

Phenylalanine dehydrogenase (L-PheDH) from Sporosarcina ureae was immobilized on DEAE-cellulose, modified initially with 2-amino-4,6-dichloro-s-triazine followed by hexamethylenediamine and glutaraldehyde. The highest activity of immobilized PheDH was determined as 95.75 U/g support with 56% retained activity. The optimum pH value of immobilized L-PheDH was shifted from pH 10.4 to 11.0. The immobilized L-PheDH showed activity variations close to the maximum value in a wider temperature range of 45-55 °C, whereas it was 40 °C for the native enzyme. The pH and the thermal stability of the immobilized L-PheDH were also better than the native enzyme. At pH 10.4 and 25 °C, K (m) values of the native and the immobilized L-PheDH were determined as K(m Phe) = 0.118, 0.063 mM and K(m NAD)(+) = 0.234, 0.128 mM, respectively. Formed NADH at the exit of packed bed reactor column was detected by the flow-injection analysis system. The conversion efficiency of the reactor was found to be 100% in the range of 5-600 μM Phe at 9 mM NAD(+) with a total flow rate of 0.1 mL/min. The reactor was used for the analyses of 30 samples each for 3 h per day. The half-life period of the reactor was 15 days.

References

Jan 1, 1994·Acta Neuropathologica·K RenkawekW W de Jong
Mar 25, 1998·Analytical Chemistry·T HuangF W Scheller
Feb 16, 2000·Journal of the American Academy of Child and Adolescent Psychiatry·P S Jensen
Dec 1, 2001·Clinica Chimica Acta; International Journal of Clinical Chemistry·Alan Clague, Andrew Thomas
Jan 20, 2004·Molecular Genetics and Metabolism·Zhaobing DingBeat Thöny
Oct 13, 2006·Biosensors & Bioelectronics·David J WeissLaura Peterson
May 18, 2007·Nutrition·Margreet van RijnFrancjan van Spronsen

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Citations

Jun 15, 2021·Langmuir : the ACS Journal of Surfaces and Colloids·Rafael Leonardo Cruz Gomes da SilvaLuciano Caseli

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