Fabrication of a Floatable Micron-Sized Enzyme Device Using Diatom Frustules.

Immobilization of enzymes has been widely reported due to their reusability, thermal stability, better storage abilities, and so on. However, there are still problems that immobilized enzymes do not have free movements to react to substrates during enzyme reactions and their enzyme activity becomes weak. Moreover, when only the porosity of support materials is focused, some problems such as enzyme distortion can negatively affect the enzyme activity. Being a solution to these problems, a new function "floatability" of enzyme devices has been discussed. A "floatable" micron-sized enzyme device was fabricated to enhance the free movements of immobilized enzymes. Diatom frustules, natural nanoporous biosilica, were used to attach papain enzyme molecules. The floatability of the frustules, evaluated by macroscopic and microscopic methods, was significantly better than that of four other SiO2 materials, such as diatomaceous earth (DE), which have been widely used to fabricate micron-sized enzyme devices. The frustules were fully suspended at 30 °C for 1 h without stirring, although they settled at room temperature. When enzyme assays were performed at room temperature, 37, and 60 °C with or without external stirring, the proposed frustule device showed the highest enzyme activity under all conditions among papain devices similarly prepared using other SiO2 materials. It was confirmed by the free papain experiments that the frustule device was active enough for enzyme reactions. Our data indicated that the high floatability of the reusable frustule device, and its large surface area, is effective in maximizing enzyme activity due to the high probability to react to substrates.

Distinguishing Antioxidant Molecules with Near-Infrared Photoluminescence of DNA-Wrapped Single-Walled Carbon Nanotubes.

In this study, two biomolecule solutions were distinguished using the capacity difference in the near-infrared photoluminescence (PL) of single-walled carbon nanotubes (SWNTs). Biosensing techniques using sensitive responses of SWNTs have been intensively studied. When a small amount of an oxidant or reductant solution was injected into the SWNT suspensions, the PL intensity of the SWNTs is significantly changed. However, distinguishing between different molecules remains challenging. In this study, we comparably injected saponin and banana solutions, which are known antioxidant chemicals, into an SWNT suspension. The SWNTs were solubilized by wrapping them with DNA molecules. The results show that 69.1 and 155.2% increases of PL intensities of SWNTs were observed after injection of 20 and 59 µg/mL saponin solutions, respectively. Subsequently, the increase in PL was saturated. With the banana solution, 18.1 and 175.4% increases in PL intensities were observed with 20 and 59 µg/mL banana solutions, respectively. Based on these results, the two antioxidant molecules could be distinguished based on the different PL responses of the SWNTs. In addition, the much higher saturated PL intensities observed with the banana solution suggests that the banana solution increased the capacity of the PL increase for the same SWNT suspension. These results provide helpful information for establishing biosensing applications of SWNTs, particularly for distinguishing chemicals.

Attachment of DNA-Wrapped Single-Walled Carbon Nanotubes (SWNTs) for a Micron-Sized Biosensor.

We fabricated a micron-sized biodevice based on the near-infrared photoluminescence (PL) response of single-walled carbon nanotubes (SWNTs). Various biosensors using the unique optical responses of SWNTs have been proposed by many research groups. Most of these employed either colloidal suspensions of dispersed SWNTs or SWNT films on flat surfaces, such as electrodes. In this study, we attached DNA-wrapped SWNTs (DNA-SWNTs) to frustule (micron-sized nanoporous biosilica) surfaces, which were purified from cultured isolated diatoms. After the injection of an oxidant and a reductant, the SWNTs on the frustules showed prominent PL responses. This suggests that the biodevice functions as a micron-sized redox sensor. Frustules can be easily suspended in aqueous solutions because of their porous structures and can easily be collected as pellets by low-speed centrifugation. Thus, the removal of unbound SWNTs and the recovery of the fabricated DNA-SWNT frustules for reuse were achieved by gentle centrifugation. Our proposal for micron-sized SWNT biodevices would be helpful for various biological applications.