Exploring and applying the substrate promiscuity of a C-glycosyltransferase in the chemo-enzymatic synthesis of bioactive C-glycosides

Bioactive natural C-glycosides are rare and chemical C-glycosylation faces challenges while enzymatic C-glycosylation catalyzed by C-glycosyltransferases provides an alternative way. However, only a small number of C-glycosyltransferases have been found, and most of the discovered C-glycosyltransferases prefer to glycosylate phenols with an acyl side chain. Here, a promiscuous C-glycosyltransferase, AbCGT, which is capable of C-glycosylating scaffolds lacking acyl groups, is identified from Aloe barbadensis. Based on the substrate promiscuity of AbCGT, 16 C-glycosides with inhibitory activity against sodium-dependent glucose transporters 2 are chemo-enzymatically synthesized. The C-glycoside 46a shows hypoglycemic activity in diabetic mice and is biosynthesized with a cumulative yield on the 3.95 g L‒1 scale. In addition, the key residues involved in the catalytic selectivity of AbCGT are explored. These findings suggest that AbCGT is a powerful tool in the synthesis of lead compounds for drug discovery and an example for engineering the catalytic selectivity of C-glycosyltransferases.

Isolation of Biofilm-Forming Bacteria from the Secondary Effluent of the Wastewater Treatment Plant and its Ability to Produce N-Acylhomoserine Lactone as Quorum Sensing Signal

Bacterial quorum sensing (QS) is one of the cell-to-cell communication systems. N-Acyl-homoserine lactones (AHLs) are the most common QS signals and responsible for biofilm formation in gram-negative bacteria. Inactivation of QS, also referred to as quorum quenching, has been regarded as a popular strategy to control membrane bioreactor (MBR) operations because biofouling in MBR wastewater treatment systems is often caused by biofilm formation. In the present study, 24 bacterial strains were isolated from the secondary effluent of a wastewater treatment plant and AHL productivity and biofilm formation were evaluated on 8 out of 24 isolates. The isolated 8 strains can produce AHLs that only possess a long (C8-C14) acyl side chain. The representative 3 strains were selected from these isolates based on the higher activity of biofilm formation. The AHL separation analysis with a thin layer chromatography clearly showed that N-octanoyl-L-homoserine lactone (C8-HSL) and N-hexanoyl-L-homoserine lactone (C6-HSL) existed in the secondary effluent sample, while no C6-HSL producing strain was isolated. C8-HSL was identified as the product of isolate No. 6. These results suggest that the secondary effluent probably contains various cell-to-cell signaling molecules derived not only from the inhabitants but also from the other microorganisms involved in the activated sludge for the biological pre-treatment. In this secondary effluent, AHL trapping techniques can be proposed as one of the acceptable strategies for the control of the QS systems because the remaining AHLs have relatively long acyl side chain and low concentrations.

Synthesis of 3-acyltetramates by side chain manipulation and their antibacterial activity

An efficient approach for the introduction of 3-acyl side chain groups onto a core tetramate system, which are suitable for further manipulation by nucleophilic displacement or Horner–Wadsworth–Emmons coupling, provides access to a diverse library of substituted tetramates related to two distinct classes of natural products, equisetin and pramanicin.