Quantitative Determination of Glycine in Aqueous Solution Using Glutamate Dehydrogenase-Immobilized Glyoxal Agarose Beads

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).

The effects of acetaldehyde, glyoxal and acetic acid on the heterogeneous reaction of nitrogen dioxide on gamma-alumina

“…The broad band from 3630 to 2780 cm À1 reflects the OH groups of CH(OH) 2 CHO. 59 The band appearing at 1661 cm À1 is due to the n(CQO) stretching vibration, 59,60 indicating the coordination of glyoxal molecules with Lewis sites through the carbonyl oxygen. The observed species indicates that gaseous glyoxal is easily converted to other surface species, which is consistent with the previous studies that have demonstrated that OHC-CHO can react with surface adsorbed water to form CH(OH) 2 CHO.…”

Ultrastable Gold–Copper Nanoclusters on Nitrogen-Doped Graphene Quantum Dots for Selective Electrochemical and Fluorescence Sensing of Glycine

“…The synthesis of AuCuNC@N-GQD was done through the GE method as follows: in a typical synthesis, 0.5 mL of AuCl 3 (∼0.17 M) was added to 6 mL of CuNC@N-GQD; the synthesis procedure of N-GQD and CuNC@N-GQD is reported elsewhere − by our group. The reaction mixture was stirred for 4 h at 65 °C until the turbid solution became clear.…”

“…These indirect methods involve inter-reactions, making the sensing procedure more complex and hence becoming time-consuming. For instance, the enzymatic sensing procedure uses the enzyme glutamate dehydrogenase (GDH) 35 to determine GLY. Here, ammonia is produced from the enzymatic reactions between GLY and GDH.…”

“…32 Thus, the quantitative sensing of GLY holds a high significance in the commercial and scientific fields of biology, medicine, and food science. Present methods of detection of GLY involve enzymes, whether it is a complex method such as liquid chromatography, 34 spectrophotometry, 35 or fluorescence assay kits, 36 and it possesses demerits such as presample preparation, experimental complexity, difficulty in attaining higher sensitivity and lower limit of detection (LOD), time consumption, and not at all costeffective, 35 whereas electrochemical (EC) and fluorescence sensing methods are relatively simpler, has higher sensitivity, low operating cost, and fast response, in which the EC method is highly meritorious because of its experimental simplicity, easily portable nature, higher sensitivity, and cost-effectiveness as compared with the fluorescence sensing. 37 In this work, we demonstrate a dual-sensing property of AuCuNC@N-GQD toward GLY via both EC and fluorescence methods.…”

Ca@Cu-CD nanoprobe for dual detection of glycine and ex vivo glycine imaging

“…7 So far, several research papers have been published on the quantitative detection of glycine, which mainly involve enzymes. 8 Such instrument-based methods, including spectrophotometry, 9 liquid chromatography, 10 fluorescence assay, 11 atomic absorption spectrometry (AAS), 12 calorimetric methods, 13 Raman spectroscopy, 14 and two-dimensional infrared spectroscopy (2D-IR), 15 possess disadvantages such as delicate sample preparation, experimental complexity, time consumption, and high cost. With the current research progress in nanotechnology, a number of nanoprobes have been designed for effective disease diagnosis at the cellular and molecular level.…”