Novel artificial metalloenzymes by in vivo incorporation of metal-binding unnatural amino acids

Artificial metalloenzymes have emerged as an attractive new approach to enantioselective catalysis. Herein, we introduce a novel strategy for preparation of artificial metalloenzymes utilizing amber stop codon suppression methodology for the in vivo incorporation of metal-binding unnatural amino acids. The resulting artificial metalloenzymes were applied in catalytic asymmetric Friedel-Crafts alkylation reactions and up to 83% ee for the product was achieved

Factors Controlling Contemporary Suspended Sediment Yield in the Caucasus Region

This paper discusses the joint impact of catchment complexity in topography, tectonics, climate, landuse patterns, and lithology on the suspended sediment yield (SSY, t km−2 year−1) in the Caucasus region using measurements from 244 gauging stations (GS). A Partial Least Square Regression (PLSR) was used to reveal the relationships between SSY and explanatory variables. Despite possible significant uncertainties on the SSY values, analysis of this database indicates clear spatial patterns of SSY in the Caucasus. Most catchments in the Lesser Caucasia and Ciscaucasia are characterized by relatively low SSY values (−2 year−1), the Greater Caucasus region generally have higher SSY values (more than 150–300 t km−2 year−1). Partial correlation analyses demonstrated that such proxies of topography as height above nearest drainage (HAND) and normalized steepness index (Ksn) tend to be among the most important ones. However, a PLSR analysis suggested that these variables’ influence is likely associated with peak ground acceleration (PGA). We also found a strong relationship between land cover types (e.g., barren areas and cropland) and SSY in different elevation zones. Nonetheless, adding more gauging stations into analyses and more refined characterizations of the catchments may reveal additional trends

Recent advances in enzymatic and chemical deracemisation of racemic compounds

Contains fulltext : 123227pos.pdf (postprint version ) (Open Access

Mutations of an NAD (P) H-dependent flavoprotein monooxygenase that influence cofactor promiscuity and enantioselectivity

AbstractThe flavoprotein monooxygenase (FPMO) from Stenotrophomonas maltophilia (SMFMO, Uniprot: B2FLR2) catalyses the asymmetric oxidation of thioethers and is unusual amongst FPMOs in its ability to use the non-phosphorylated cofactor NADH, as well as NADPH, for the reduction of the FAD coenzyme. In order to explore the basis for cofactor promiscuity, structure-guided mutation of two residues in the cofactor binding site, Gln193 and His194, in SMFMO were performed in an attempt to imitate the cofactor binding site of the NADPH-dependent FMO from Methylophaga aminisulfidivorans sp. SK1 (mFMO), in which structurally homologous residues Arg234 and Thr235 bind the NADPH 2′-ribose phosphate. Mutation of His194 to threonine proved most significant, with a switch in specificity from NADH to NADPH [(kcat/Km NADH)/kcat/Km NADPH) from 1.5:1 to 1:3.5, mostly as a result of a reduced Km for NADPH of approximately sevenfold in the His194Thr mutant. The structure of the Gln193Arg/His194Thr mutant revealed no substantial changes in the backbone of the enzyme or orientation of side chains resulting from mutation. Mutation of Phe52, in the vicinity of FAD, and which in mFMO is an asparagine thought to be responsible for flavin hydroperoxide stabilisation, is, in SMFMO, a determinant of enantioselectivity in sulfoxidation. Mutation of Phe52 to valine resulted in a mutant that transformed para-tolyl methyl sulfide into the (S)-sulfoxide with 32% e.e., compared to 25% (R)- for the wild type. These results shed further light both on the cofactor specificity of FPMOs, and their determinants of enantioselectivity, with a view to informing engineering studies of FPMOs in the future

A rational quest for selectivity through precise ligand-positioning in the tandem DNA-catalysed Friedel-Crafts alkylation/asymmetric protonation

International audienceCovalent anchorage of a metallic co-factor to a DNA-based architecture is merely the only way to ensure an accurate positioning of a catalytic site within the chiral micro-environment offered by the DNA double helix. Ultimately, it also allows a fine-tuning of the catalytic pocket through simple synthetic modifications of the DNA sequence. Here, we report highly selective copper(II)-catalysed asymmetric Friedel-Crafts conjugate addition/ enantioselective protonation, which is due to a careful positioning of a bipyridine ligand within a DNA framework. Most importantly, this study unveils specific structural features that account for an optimal chirality transfer from the duplex to the Friedel-Crafts adducts