Tetramerization of GH2 ß-Glucuronidases is Essential for Catalyzing the Hydrolysis of the Large Substrate Glycyrrhizin.

In this study, structural analysis was employed to identify three hotspot residues that contribute most to the tetramer formation of a glycoside hydrolase family 2 (GH2) ß-glucuronidase (GUS) from Aspergillus oryzae Li-3. Single-point mutation at these sites completely disrupted the tetramer structure and abolished the glycyrrhizin (GL)-hydrolyzing activity. Then, the W522A dimer was refactored into a tetramer by disulfide bonding, and partial GL activity was restored. Further saturated mutation showed a strong correlation between the GL activity of the mutants and their tetramer ratios. Molecular simulations were employed to illustrate the critical role of the tetramer interface in maintaining a functional active-site structure. The three highly conserved tetramer-forming residues were finally applied to two other GH2 GUSs for tetramer dissociation and demonstrated the significance of the homotetramerization for GL-hydrolyzing activity of GH2 GUSs. This study lays foundation for engineering GL-hydrolyzing GUSs at the quaternary structure level for function regulations.

Isolated single quantum dot emitters in all-epitaxial microcavities.

Data are presented on a fabrication approach that places an isolated single quantum dot at the center of a semiconductor microcavity. The microcavity is based on an all-epitaxial mesa-confined design that is mechanically robust and provides the thermal dissipation needed for a single photon source device technology. Microphotoluminescence is used to reveal single quantum dot emission with the essential optical properties of single quantum emitters.