Sulfonium Acids Loaded onto an Unusual Thiotemplate Assembly Line Construct the Cyclopropanol Warhead of a Burkholderia Virulence Factor

Pathogenic bacteria of the Burkholderia pseudomallei group cause severe infectious diseases such as glanders and melioidosis. Malleicyprols were identified as important bacterial virulence factors, yet the biosynthetic origin of their cyclopropanol warhead has remained enigmatic. By a combination of mutational analysis and metabolomics we found that sulfonium acids, dimethylsulfoniumpropionate (DMSP) and gonyol, known as osmolytes and as crucial components in the global organosulfur cycle, are key intermediates en route to the cyclopropanol unit. Functional genetics and in vitro analyses uncover a specialized pathway to DMSP involving a rare prokaryotic SET-domain methyltransferase for a cryptic methylation, and show that DMSP is loaded onto the NRPS-PKS hybrid assembly line by an adenylation domain dedicated to zwitterionic starter units. Then, the megasynthase transforms DMSP into gonyol, as demonstrated by heterologous pathway reconstitution in E. coli. © 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA

Simultaneous Production of Psilocybin and a Cocktail of β‐Carboline Monoamine Oxidase Inhibitors in “Magic” Mushrooms

The psychotropic effects of Psilocybe “magic” mushrooms are caused by the l ‐tryptophan‐derived alkaloid psilocybin. Despite their significance, the secondary metabolome of these fungi is poorly understood in general. Our analysis of four Psilocybe species identified harmane, harmine, and a range of other l ‐tryptophan‐derived β‐carbolines as their natural products, which was confirmed by 1D and 2D NMR spectroscopy. Stable‐isotope labeling with 13 C 11 ‐ l ‐tryptophan verified the β‐carbolines as biosynthetic products of these fungi. In addition, MALDI‐MS imaging showed that β‐carbolines accumulate toward the hyphal apices. As potent inhibitors of monoamine oxidases, β‐carbolines are neuroactive compounds and interfere with psilocybin degradation. Therefore, our findings represent an unprecedented scenario of natural product pathways that diverge from the same building block and produce dissimilar compounds, yet contribute directly or indirectly to the same pharmacological effects

Cyclopropanol Warhead in Malleicyprol Confers Virulence of Human- and Animal-Pathogenic Burkholderia Species

Burkholderia species such as B. mallei and B. pseudomallei are bacterial pathogens causing fatal infections in humans and animals (glanders and melioidosis), yet knowledge on their virulence factors is limited. While pathogenic effects have been linked to a highly conserved gene locus (bur/mal) in the B. mallei group, the metabolite associated to the encoded polyketide synthase, burkholderic acid (syn. malleilactone), could not explain the observed phenotypes. By metabolic profiling and molecular network analyses of the model organism B. thailandensis, the primary products of the cryptic pathway were identified as unusual cyclopropanol-substituted polyketides. First, sulfomalleicyprols were identified as inactive precursors of burkholderic acid. Furthermore, a highly reactive upstream metabolite, malleicyprol, was discovered and obtained in two stabilized forms. Cell-based assays and a nematode infection model showed that the rare natural product confers cytotoxicity and virulence

Two-body wear rate of CAD/CAM resin blocks and their enamel antagonists

STATEMENT OF PROBLEM: Computer-aided design and computer-aided manufacturing (CAD/CAM) resins exhibit good mechanical properties and can be used as long-term restorations. The wear rate of such resins and their enamel antagonists is unknown. PURPOSE: The purpose of this study was to test and compare the 2-body wear rate of CAD/CAM resin blocks. MATERIAL AND METHODS: Wear specimens (N=42, n=6) were made from 5 CAD/CAM resins: ZENO PMMA (ZP), artBloc Temp (AT), Telio CAD (TC), Blanc High-class (HC), CAD-Temp (CT); 1 manually polymerized resin: Integral esthetic press (negative control group, IEP); and 1 glass-ceramic: VITA Mark II (positive control group, VM2). The specimens for the wear resistance were aged in a thermomechanical loading machine (49 N, 1.67 Hz, 5/50°C) with human enamel antagonists. The material loss of all specimens before, during, and after aging was evaluated with a 3DS profilometer. The measured material loss data of all tested groups were statistically evaluated with linear mixed model analysis (a=.05). RESULTS: Manually polymerized resin showed significantly higher material wear (P<.001) than all other tested groups. Glass-ceramic showed significantly lower wear values (P<.001) than CAD/CAM resins ZP, AT, HC, CT, and IES. CAD/CAM resin TC was not significantly different from the positive control group. Glass-ceramic showed the highest enamel wear values (P<.001) of all tested resins. No differences were found in the enamel wear among all resins. The glass-ceramic group showed damage in the form of cracks on the worn enamel surface in 50% of specimens. CONCLUSIONS: CAD/CAM resins showed lower wear rates than those conventionally polymerized. Only one CAD/CAM resin, TC, presented material wear values comparable with glass-ceramic. The tested glass-ceramic developed cracks in the enamel antagonist and showed the highest enamel wear values of all other tested groups

Two-body wear of monolithic, veneered and glazed zirconia and their corresponding enamel antagonists

Abstract Objective. This study tested whether the two-body wear of monolithic zirconia and their corresponding enamel antagonists was higher compared to monolithic alloy and veneered zirconia. Materials and methods. Cylindrical specimens (N = 36, n = 6) were prepared out of (A) veneered zirconia (VZ), (B) glazed zirconia using a glaze ceramic (GZC), (C) glazed zirconia using a glaze spray (GZS), (D) manually polished monolithic zirconia (MAZ), (E) mechanically polished monolithic zirconia (MEZ) and (F) monolithic base alloy (control group, MA). Wear tests were performed in a chewing simulator (49 N, 1.7 Hz, 5°C/50°C) with enamel antagonists. The wear analysis was performed using a 3D profilometer before and after 120,000, 240,000, 640,000 and 1,200,000 masticatory cycles. SEM images were used for evaluating wear qualitatively. The longitudinal results were analysed using linear mixed models (α = 0.05). Results. Materials (p < 0.001) and number of masticatory cycles (p < 0.001) had a significant effect on the wear level. The least enamel antagonist wear was observed for MAZ and MEZ (27.3 ± 15.2, 28 ± 11.1 μm, respectively). GZC (118 ± 30.9 μm) showed the highest wear of enamel antagonists. The highest wear rate in the material was observed in GZS (91.3 ± 38.6 μm). While in the groups of MA, VZ, GZC and GZS 50% of the specimens developed cracks in enamel, it was 100% in MAZ and MEZ groups. Conclusion. Polished monolithic zirconia showed lower wear rate on enamel antagonists as well as within the material itself but developed higher rates of enamel cracks