A highly selective C-rhamnosyltransferase from Viola tricolor and insights into its mechanisms.

C-Glycosides are important natural products with various bioactivities. In plant biosynthetic pathways, the C-glycosylation step is usually catalyzed by C-glycosyltransferases (CGTs), and most of them prefer to accept uridine 5'-diphosphate glucose (UDP-Glc) as sugar donor. No CGTs favoring UDP-rhamnose (UDP-Rha) as sugar donor has been reported, thus far. Herein, we report the first selective C-rhamnosyltransferase VtCGTc from the medicinal plant Viola tricolor. VtCGTc could efficiently catalyze C-rhamnosylation of 2-hydroxynaringenin 3-C-glucoside, and exhibited high selectivity towards UDP-Rha. Mechanisms for the sugar donor selectivity of VtCGTc were investigated by molecular dynamics (MD) simulations and molecular mechanics with generalized Born and surface area solvation (MM/GBSA) binding free energy calculations. Val144 played a vital role in recognizing UDP-Rha, and the V144T mutant could efficiently utilize UDP-Glc. This work provides a new and efficient approach to prepare flavonoid C-rhamnosides such as violanthin and iso-violanthin.

Hierarchical Nanoscale Structuring of Solution-Processed 2D van der Waals Networks for Wafer-Scale, Stretchable Electronics.

Two-dimensional (2D) semiconductors are promising for next-generation electronics that are lightweight, flexible, and stretchable. Achieving stretchability with suppressed crack formation, however, is still difficult without introducing lithographically etched micropatterns, which significantly reduces active device areas. Herein, we report a solution-based hierarchical structuring to create stretchable semiconducting films that are continuous over wafer-scale areas via self-assembly of two-dimensional nanosheets. Electrochemically exfoliated MoS2 nanosheets with large lateral sizes (∼1 µm) are first assembled into a uniform film on a prestrained thermoplastic substrate, followed by strain relief of the substrate to create nanoscale wrinkles. Subsequent strain-relief cycles with the presence of soluble polymer films produce hierarchical wrinkles with multigenerational structures. Stretchable MoS2 films are then realized by curing an elastomer directly on the wrinkled surface and dissolving the thermoplastic. Three-generation hierarchical MoS2 wrinkles are resistant to cracking up to nearly 100% substrate stretching and achieve drastically enhanced photoresponsivity compared to the flat counterpart over the visible and NIR regimes, while the flat MoS2 film is beneficial in creating strain sensors because of its strain-dependent electrical response.

GuRhaGT, a highly specific saponin 2''-O-rhamnosyltransferase from Glycyrrhiza uralensis.

A highly regio- and donor-specific 2''-O-rhamnosyltransferase GuRhaGT was characterised from the medicinal plant Glycyrrhiza uralensis. GuRhaGT could efficiently catalyse rhamnosylation at 2''-OH of the C-3 glycosyl moiety of triterpenoid saponins.

Dissection of the general two-step di-C-glycosylation pathway for the biosynthesis of (iso)schaftosides in higher plants.

Schaftoside and isoschaftoside are bioactive natural products widely distributed in higher plants including cereal crops and medicinal herbs. Their biosynthesis may be related with plant defense. However, little is known on the glycosylation biosynthetic pathway of these flavonoid di-C-glycosides with different sugar residues. Herein, we report that the biosynthesis of (iso)schaftosides is sequentially catalyzed by two C-glycosyltransferases (CGTs), i.e., CGTa for C-glucosylation of the 2-hydroxyflavanone aglycone and CGTb for C-arabinosylation of the mono-C-glucoside. The two enzymes of the same plant exhibit high homology but remarkably different sugar acceptor and donor selectivities. A total of 14 CGTa and CGTb enzymes were cloned and characterized from seven dicot and monocot plants, including Scutellaria baicalensis, Glycyrrhiza uralensis, Oryza sativa ssp. japonica, and Zea mays, and the in vivo functions for three enzymes were verified by RNA interference and overexpression. Through transcriptome analysis, we found homologous genes in 119 other plants, indicating this pathway is general for the biosynthesis of (iso)schaftosides. Furthermore, we resolved the crystal structures of five CGTs and realized the functional switch of SbCGTb to SbCGTa by structural analysis and mutagenesis of key amino acids. The CGT enzymes discovered in this paper allow efficient synthesis of (iso)schaftosides, and the general glycosylation pathway presents a platform to study the chemical defense mechanisms of higher plants.