The conversion of fumaric acid into L-malate by fumarase immobilized on silanized nanostructures was analyzed experimentally. The enzyme was bound to the silanized nanostructures. We carried out scanning electron microscopy (SEM), fourier transform infrared spectroscopy (FTIR) analysis, zeta size analysis and surface area calculation for the characterization of the nanostructures. The effect of initial enzyme concentration and pH on immobilization procedure were investigated and the change of Michaelis-Menten constants (K m and V max ) with immobilization was examined. The change in the storage stability of the enzyme by immobilization was also investigated. The stability of the immobilized enzyme was very good. We observed that the fumarase was bound to silanized nanostructures [p(HEMA)-3-MTES] in much greater amounts. We have compared the activities of free fumarase and immobilized fumarase and we have observed a significant increase in the activity of the fumarase after immobilization for L-malate production. Moreover, we came to the conclusion that this activity can be better preserved for 30 days compared to free fumarase.
Folic acid, which provides the transfer of single carbon atoms in synthesis reactions and metabolic cycles in metabolism, is very important for metabolism. Folic acid also plays an important role in nucleotide synthesis and methylation reactions. There are many disorders caused by defective folic acid metabolism and lack of folic acid. Today, innovative, cost-effective methods are needed to develop folic acid determination methods. The main objective of this study is the development of surface-printed carbon electrodes (SPCE) modified with folic acid imprinted nanostructures (FA-Imp-poly(MPTS-rGO-co-NAT), which will be used for the first time for folic acid determination in commercially human blood serum. For this purpose, the synthesis of nanostructures has been carried out and characterized by FTIR, SEM-EDS, and AFM. Then, a new chemically modified nanosensor was fabricated for the determination of folic acid using folic acid imprinted nanostructures. Differential pulse voltammetry (DPV) and circular voltammetry (CV) methods were used as electrochemical methods in the FA-imprinted-nanosensor studies. Measurements in differential pulse voltammetry were performed at an application speed of 0.005 volts per second in the potential range of −0.4 to 0.6 volts. As a result of the circular voltammetric method, an idea about the surface was obtained with the voltammograms obtained. The detection limit (LOD) of the developed FA-imprinted-nanosensor was 7.54 ng/mL and the determination limit (LOQ) was 25.14 ng/mL. FA analytical (10 and 20 ng/mL) was added to commercial synthetic serum samples by the standard adding method and RSD values of 0.092% and 0.734% were found in the DPV technique and measurements respectively. This manuscript demonstrated a novel, simple, selective, and rapid FA-imprinted-nanosensor for determining the FA in the biological samples.
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