In this study, we prepared graphene oxide (GO)/ZIF-67 nanocomposites. Therefore, GO/ZIF-67 nanocomposites were used as a modifier on a screen-printed electrode (GO/ZIF-67/SPE) for studying the electrochemical behavior of epinine in phosphate buffer saline (PBS) at pH 7.0 with voltammetry techniques. The GO/ZIF-67/SPE showed greater electrocatalytic activities than the bare SPE. As a result, the GO/ZIF-67/SPE was utilized for additional electrochemical examinations. The epinine concentration determination was in the range 9.0 × 10⁻⁸ M to 5.0 × 10⁻⁴ M, and the limit of detection (LOD) as well as the limit of quantification (LOQ) equaled 2.0 and 6.6 nM, respectively. From the scan rate study, the oxidation of epinine was found to be diffusion-controlled, and the simultaneous detection of epinine and dobutamine were well achieved with the differential pulse voltammetric (DPV) technique. Moreover, the stability and reproducibility of epinine at the GO/ZIF-67/SPE was studied, and the use of the GO/ZIF-67/SPE to detect epinine and dobutamine in real samples was furthermore successfully demonstrated.

In most bioluminescence systems the oxidation of luciferin and production of light is catalyzed by luciferases. Protein engineering studies have shown that thermostable proteins from thermophilic organisms have a higher frequency of Arg, especially in exposed states. To further clarify the arginine saturation effect on thermostability of firefly luciferase, some of hydrophobic solvent-exposed residues in Lampyris turkestanicus luciferase are changed to arginine. All of these residues are located at the surface loops of L.turkestanicus luciferase. Starting with a luciferase mutant (E³⁵⁴Q/Arg³⁵⁶), single mutation (-Q³⁵R, -I¹⁸²R, -I²³²R and -L³⁰⁰R), double mutation (-Q³⁵R/I²³²R) and triple mutation (-Q³⁵R/I²³²R/I¹⁸²R) are added. Bioluminescence emission spectra indicate that substitution of Arg by these residues, do not effect on the maximum wavelength of emission spectrum. It should be noted, introduction of double mutation (-Q³⁵R/I²³²R) and triple mutation (-Q³⁵R/I²³²R/I¹⁸²R) were kept specific activity of firefly luciferase. By addition of positively charged residue, some specific mutations (-I²³²R, -Q³⁵R/I²³²R and -Q³⁵R/I²³²R/I¹⁸²R) showed that optimum temperature of activity was increased to 40°C which are 12 and 15°C higher than E³⁵⁴Q/Arg³⁵⁶ and wild-type luciferases, respectively. Also, after 40 min incubation of enzymes at 40°C, the relative remaining activity of wild type was only 5%, whereas for -I²³²R, -Q³⁵R/I²³²R and -Q³⁵R/I²³²R/I¹⁸²R was 60, 80 and 80% of original activity, respectively.

The improvement in the enzyme activity of Aspergillus flavus urate oxidase (Uox) was attained by immobilizing it on the surface of a Ni-based magnetic metal–organic framework (NimMOF) nanomaterial; physicochemical properties of NimMOF and its application as an enzyme stabilizing support were evaluated, which revealed a significant improvement in its stability upon immobilization on NimMOF (Uox@NimMOF). It was affirmed that while the free Uox enzyme lost almost all of its activity at ~40–45 °C, the immobilized Uox@NimMOF retained around 60% of its original activity, even retaining significant activity at 70 °C. The activation energy (Ea) of the enzyme was calculated to be ~58.81 kJ mol⁻¹ after stabilization, which is approximately half of the naked Uox enzyme. Furthermore, the external spectroscopy showed that the MOF nanomaterials can be coated by hydrophobic areas of the Uox enzyme, and the immobilized enzyme was active over a broad range of pH and temperatures, which bodes well for the thermal and long-term stability of the immobilized Uox on NimMOF.