Abstract
In this study, an enzymatic procedure for the determination of glycine (Gly) was developed by using a column containing immobilized glutamate dehydrogenase (GDH) on glyoxal agarose beads. Ammonia is produced from the enzymatic reactions between Gly and GDH with NAD+ in phosphate buffer medium. The indophenol blue method was used for ammonia detection based on the spectrophotometric measurements of blue-colored product absorbing at 640 nm. The calibration graph is linear in the range of 0.1–10 mM of Gly concentrations. The effect of pH, temperature, and time interval was studied to find column stability, and also the interference effects of other amino acids was investigated. The interaction between GDH and glyoxal agarose beads was analyzed by Fourier transform infrared (FTIR) spectroscopy. The morphology of the immobilized and non-immobilized agarose beads were characterized by atomic force microscopy (AFM).
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Mitic, S. S., Pavlovic, A. N., Tosic, S. B., Arsic, B. B., & Sunaric, S. M. (2009). Journal of Analytical Chemistry, 64, 683–689.
Li, Q. Z., Huang, Q. X., Li, S. C., Yang, M. Z., & Rao, B. (2012). Korean Journal of Physiology and Pharmacology, 16, 355–360.
Tang, Y. B., Teng, L., Sun, F., Wang, X. L., Peng, L., Cui, Y. Y., et al. (2012). Journal of Chromatography B, 905, 61–66.
Culea, M., Horj, E., Iordache, A., & Cozar, O. (2011). Asian Journal of Chemistry, 23, 4279–4281.
Wilson, S. F., James, C. A., Zhu, X., Davis, M. T., & Rose, M. J. (2011). Journal of Pharmaceutical and Biomedical, 56, 315–323.
Pakhomova, O. A., Korenman, Y. I., Mokshina, N. Y., & Niftaliev, S. I. (2010). Russian Journal of Applied Chemistry, 83, 1940–1943.
Zyablov, A. N., Kalach, A. V., Zhibrova, Y. A., Selemenev, V. F., & Dyakonova, O. V. J. (2010). Analytical Chemistry, 65, 91–93.
Tuma, P., Soukupova, M., Samcova, E., & Stulık, K. (2009). Electrophoresis, 30, 3436–3441.
Marrubini, G., Caccialanza, G., & Massolini, G. (2008). Journal of Pharmaceutical and Biomedical, 47, 716–722.
Osuji, G. O., Konan, J., & M’mbijjewe, G. (2004). Applied Biochemistry and Biotechnology, 119, 209–228.
Blasi, L., Longo, L., Vasapollo, G., Cingolani, R., Rinaldi, R., Rizzello, T., et al. (2005). Enzyme and Microbial Technology, 36, 818–823.
Petach, H. H., & Driscoll, J. (1994). Biotechnology and Bioengineering, 44, 1018–1022.
Cordek, J., Wang, X., & Tan, W. (1999). Analytical Chemistry, 71, 1529–1533.
O’Connor, K. C., Schütz, H. J., & Bailey, J. E. (1989). Biotechnology and Bioengineering, 33, 896–905.
Stalikas, C. D., Karayannis, M. I., & Tzouwara-Karayanni, S. (1994). Talanta, 41, 1561–1567.
Gambhir, A., Gerard, M., Mulchandani, A. K., & Malhotra, B. D. (2001). Applied Biochemistry and Biotechnology, 96, 249–257.
Longo, L., Vasapollo, G., Guascito, M. R., & Malitesta, C. (2006). Analytical and Bioanalytical Chemistry, 385, 146–152.
Doong, R., & Shih, H. (2006). Biosensors and Bioelectronics, 22, 185–191.
Bolivar, J. M., Cava, F., Mateo, C., Rocha-Martin, J., Guisan, J. M., Berenguer, J., et al. (2008). Applied Microbiology and Biotechnology, 80, 49–58.
Mateo, C., Palomo, J. M., Fuentes, M., Betancor, L., Grazu, V., Lopez-Gallego, F., et al. (2006). Enzyme and Microbial Technology, 39, 274–280.
Manrich, A., Komesu, A., Adriano, W. S., Tardioli, P. W., & Giordano, R. L. C. (2010). Applied Biochemistry and Biotechnology, 161, 455–467.
Tzollas, N. M., Zachariadis, G. A., Anthemidis, A. N., & Stratis, J. A. (2010). International Journal of Environmental Analytical Chemistry, 90, 115–126.
Yaqoob, M., & Nabi, A. (2001). Talanta, 55, 1181–1186.
Villas-Boas, S. G., Akesson, M., & Nielsen, J. (2005). FEMS Yeast Research, 5, 703–709.
Peterson, P. E., & Smith, T. J. (1999). Structure, 7, 769–782.
Pompa, P. P., Blasi, L., Longo, L., Cingolani, R., Ciccarella, G., Vasapollo, G., Rinaldi, R., Rizzello, A., Storelli, C., & Maffia, M. (2003). Physical Review E, 67(4 Pt 1), 041902.
Kwinta, J., Bartoszewicz, K., & Bielawski, W. (2002). Acta Physiologiae Plantarum, 24, 279–283.
Ma, K., Robb, F. T., & Adams, M. W. (1994). Applied and Environmental Microbiology, 60, 562–568.
Gannon, M. C., Nuttall, J. A., & Nuttall, F. Q. (2002). The American Journal of Clinical Nutrition, 76, 1302–1307.
Ditscheid, B., Fünfstück, D. R., Busch, M., Schubert, R., Gerth, J., & Jahreis, G. (2005). European Journal of Clinical Nutrition, 59, 768–775.
Brand, H. S., Jörning, G. G. A., Chamuleau, R. A. F. M., & Abraham-Inpijn, L. (1997). Clinica Chimica Acta, 264, 37–47.
Kaspar, H., Dettmer, K., Gronwald, W., & Oefner, P. J. (2008). Journal of Chromatography B, 870, 222–232.
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This work was financially supported by Sakarya University Commission of Scientific Research Projects (project number 2012-02-04-041).
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Keskin, S.Y., Keskin, C.S. Quantitative Determination of Glycine in Aqueous Solution Using Glutamate Dehydrogenase-Immobilized Glyoxal Agarose Beads. Appl Biochem Biotechnol 172, 289–297 (2014). https://doi.org/10.1007/s12010-013-0543-1
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DOI: https://doi.org/10.1007/s12010-013-0543-1