'Urease immobilized single-kit' for sensing of thiourea-glucose pair employing fluorescence 'Turn off - Turn on' and as an efficient sorbent for selective sample cleanup of thiourea.

The tenfold lowering in binding energy for TU-Tyrosine in immobilized urease (Kb: 4.7 × 103) with respect to the native enzyme (Kb: 6.5 × 104) begets easy desorption of thiourea (TU) by glucose (GL) with an eventual formation of a more strong TU- GL adduct; that rejuvenates the kit-material ready for the subsequent cycle(s). The sorption-desorption heeds fluorescence turn-off and turn-on in DCM for selective sensing of TU- GL pair at their respective linear range of concentration 2.5-26.1 ppm and 2.36-11.57 ppm. The process was found to be static (KSV ≥ 2.25 × 103 L mol-1), exothermic (ΔH: -0.08 kJ mol-1), spontaneous (ΔG: -21.1 kJ mol-1) and marginally entropy gaining (ΔS: 0.07 kJ mol-1 K-1). The 'bulk material' (200 ± 20 µm) brilliantly preconcentrates TU with an enrichment factor of 106.2 after its selective extraction at near-neutral pH from a large volume sample (800 mL) of low concentration (30 ppm). A very dilute solution (0.05 mmol L-1) of GL at minimum volume (6 mL) acts as a stripping agent and provides a longer life (200 cycles with good extraction efficiency) to the material. The method was found to be efficient in the analysis of fruit juice as a real sample.

Exuberant Immobilization of Urease on an Inorganic SiO2 Support Enhances the Enzymatic Activities by 3-fold for Perennial Utilization.

Urease has been covalently immobilized on a 3-D networking silica gel (SG) using dimethyldichlorosilane (DMDCS) as second generation silane coupling reagent and m-nitroaniline as linker component in a robust methodology and subsequently characterized as [{Si(OSi)4(H2O)0.05}205.2] n=4{OSi(CH3)2-NH-C6H4-N═N-urease}·282.5H2O (molecular mass 263 445 g or 263.4 kDa). Selective coupling of tyrosine residue with an identifiable m-nitroaniline modified SG unit prevents enzyme-enzyme cross-linking leading to enhancement of enzymatic activity. The material worked at room temperature and its activity (luminescent and ammonia releasing efficiency) was enhanced by 3-fold (for both synthetic and real sample) compared to native enzyme values at neutral pH. Up to 30 days and 30 cycles, this 3-fold activity remains as such but reduces gradually to native enzyme level after 60 days and 60 cycles of reuse.