Hydrolytic enzymes are a topic of continual study and improvement due to their industrial impact and biological implications; however, the ability to measure the activity of these enzymes, especially in high-throughput assays, is limited to an established, few enzymes and often involves the measurement of secondary byproducts or the design of a complex degradation probe. Herein, a versatile single-walled carbon nanotube (SWNT)-based biosensor that is straightforward to produce and measure is described. The hydrolytic enzyme substrate is rendered as an amphiphilic polymer, which is then used to solubilize the hydrophobic nanotubes. When the target enzyme degrades the wrapping, the SWNT fluorescent signal is quenched due to increased solvent accessibility and aggregation, allowing quantitative measurement of hydrolytic enzyme activity. Using (6,5) chiral SWNT suspended with polypeptides and polysaccharides, turnover frequencies are estimated for cellulase, pectinase, and bacterial protease. Responses are recorded for concentrations as low as 5 fM using a well-characterized protease, Proteinase K. An established trypsin-based plate reader assay is used to compare this nanotube probe assay with standard techniques. Furthermore, the effect of freeze-thaw cycles and elevated temperature on enzyme activity is measured, suggesting freezing to have minimal impact even after 10 cycles and heating to be detrimental above 60 °C. Finally, rapid optimization of enzyme operating conditions is demonstrated by generating a response surface of cellulase activity with respect to temperature and pH to determine optimal conditions within 2 h of serial scans.
We report the electrical transport properties of conducting / insulator / conducting heterostructures by studying current -voltage IV curves at room temperature. The measurements were obtained on tunnel junctions with different areas (900 µm 2 , 400 µm 2 and 100µm 2 ) using a conducting atomic force microscope. Trilayers with GdBa 2 Cu 3 O 7 (GBCO) as the bottom electrode, SrTiO 3 or BaTiO 3 (thicknesses between 1.6 nm and 4 nm) as the insulator barrier, and GBCO or Nb as the top electrode were grown by DC sputtering on (100) SrTiO 3 substrates For SrTiO 3 and BaTiO 3 barriers, asymmetric IV curves at positive and negative polarization can be obtained using electrodes with different work function. In addition, hysteretic IV curves are obtained for BaTiO 3 barriers, which can be ascribed to a combined effect of the FE reversal switching polarization and an oxygen vacancy migration. For GBCO/ BaTiO 3 / GBCO heterostructures, the IV curves correspond to that expected for asymmetric interfaces, which indicates that the disorder affects differently the properties at the bottom and top interfaces. Our results show the role of the interface disorder on the electrical transport of conducting/ insulator/ conduction heterostructures, which is relevant for different applications, going from resistive switching memories (at room temperature) to Josephson junctions (at low temperatures).
The transport properties of ultra-thin SrTiO(3) (STO) layers grown over YBa(2)Cu(3)O(7) electrodes were studied by conductive atomic force microscopy at the nano-scale. A very good control of the barrier thickness was achieved during the deposition process. A phenomenological approach was used to obtain critical parameters regarding the structural and electrical properties of the system. The STO layers present an energy barrier of 0.9 eV and an attenuation length of 0.23 nm, indicating very good insulating properties for the development of high-quality Josephson junctions.
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