Efficient Portable Urea Biosensor Based on Urease Immobilized Membrane for Monitoring of Physiological Fluids

Numerous studies have addressed the utilization of glutaraldehyde (GA) as a homobifunctional cross-linker. However, its applicability has been impeded due to several issues, including the tendency of GA molecules to undergo polymerization. Herein, a portable urea biosensor was developed for the real-time monitoring of the flow of physiological fluids; this was achieved by using disuccinimidyl cross-linker-based urease immobilization. Urease was immobilized on a porous polytetrafluoroethylene (PTFE) solid support using different disuccinimidyl cross-linkers, namely disuccinimidyl glutarate (DSG), disuccinimidyl suberate (DSS) and bis-N-succinimidyl-(pentaethylene glycol) ester (BS(PEG)5). A urease activity test revealed that DSS exhibited the highest urease immobilizing efficiency, whereas FT-IR analysis confirmed that urease was immobilized on the PTFE membrane via DSS cross-linking. The membrane was inserted in a polydimethylsiloxane (PDMS) fluidic chamber that generated an electrochemical signal in the presence of a flowing fluid containing urea. Urea samples were allowed to flow into the urea biosensor (1.0 mL/min) and the signal was measured using chronoamperometry. The sensitivity of the DSS urea biosensor was the highest of all the trialed biosensors and was found to be superior to the more commonly used GA cross-linker. To simulate real-time monitoring in a human patient, flowing urea-spiked human serum was measured and the effective urease immobilization of the DSS urea biosensor was confirmed. The repeatability and interference of the urea biosensor were suitable for monitoring urea concentrations typically found in human patients.

Synthesis of 1,4-dihydropyrimidines with immobilized urease: effect of method immobilization on magnetic supports

The effect of the urease immobilization method was studied on magnetic supports for the Biginelli/Hantzsch reaction. For this purpose, Fe

Effect of Cultivation Time and Medium Condition in Production of Bacterial Cellulose Nanofiber for Urease Immobilization

A new nanoporous biomatrix originated from bacterial resources has been chosen for urease immobilization. Urease has been immobilized on synthesized bacterial cellulose nanofiber since this enzyme has a key role in nitrogen metabolism.Gluconacetobacter xylinumATCC 10245 has been cultivated for synthesis of a nanofiber with the diameter of 30–70 nm. Different cultivation processes in the aspect of time and cultivation medium conditions were chosen to study the performance of immobilized enzyme on four types of bacterial cellulose nanofibers (BCNs). Urease immobilization into the nanofiber has been done in two steps: enzyme adsorption and glutaraldehyde cross-linking. The results showed that the immobilized enzymes were relatively active and highly stable compared to the control samples of free enzymes. Optimum pH was obtained 6.5 and 7 for different synthesized BCNs, while the optimum temperature for immobilized urease was 50°C. Finding of the current experiment illustrated that the immobilized enzyme in optimum condition lost its initial activity by 41% after 15 weeks.