A combined computational and experimental investigation of the [2Fe–2S] cluster in biotin synthase

Biotin synthase was the first example of what is now regarded as a distinctive enzyme class within the radical S-adenosylmethionine superfamily, the members of which use Fe/S clusters as the sulphur source in radical sulphur insertion reactions. The crystal structure showed that this enzyme contains a [2Fe–2S] cluster with a highly unusual arginine ligand, besides three normal cysteine ligands. However, the crystal structure is at such a low resolution that neither the exact coordination mode nor the role of this exceptional ligand has been elucidated yet, although it has been shown that it is not essential for enzyme activity. We have used quantum refinement of the crystal structure and combined quantum mechanical and molecular mechanical calculations to explore possible coordination modes and their influences on cluster properties. The investigations show that the protonation state of the arginine ligand has little influence on cluster geometry, so even a positively charged guanidinium moiety would be in close proximity to the iron atom. Nevertheless, the crystallised enzyme most probably contains a deprotonated (neutral) arginine coordinating via the NH group. Furthermore, the Fe···Fe distance seems to be independent of the coordination mode and is in perfect agreement with distances in other structurally characterised [2Fe–2S] clusters. The exceptionally large Fe···Fe distance found in the crystal structure could not be reproduced

Molecularly imprinted polymer nanomaterials and nanocomposites by controlled/living radical polymerization.

International audienceSince the pioneering work of Wulff and Mosbach more than 30 years ago, molecular imprinting of synthetic polymers has emerged as a robust and convenient way for synthesizing polymeric receptor materials bearing specific recognition sites for target molecules. The resulting materials, molecularly imprinted polymers (MIPs), are therefore commonly referred to as ‘plastic antibodies’. They are obtained by polymerizing a scaffold around a target, or a derivate thereof, which acts as a molecular template. MIPs have been successfully applied in many areas including affinity separation, immunoassays, chemical sensing, solid-phase extraction, drug delivery, cell and tissue imaging, direct synthesis and catalysis. In terms of affinity and selectivity, MIPs are on a par with biological receptors like antibodies, and this is accompanied by a superior chemical and physical stability, compatibility with organic media, reusability, easy engineering and low cost. These advantages represent the main reasons for the wide interest raised around molecularly imprinted materials. Mainly produced by free radical polymerization (FRP) of vinyl monomers, MIPs have also taken advantage of the introduction of controlled/living radical polymerization (CRP) techniques, which have literally transformed polymer chemistry over the last decade. This review describes the advantages arising from the use of CRP in synthesizing MIPs, both in terms of sheer binding properties as well as for their remarkabl

Solid-phase extraction of betanin and isobetanin from beetroot extracts using a dipicolinic acid molecularly imprinted polymer.

International audienceBetanin is a natural pigment with significant antioxidant and biological activities currently used as food colorant. The isolation of betanin is problematic due to its instability. In this work, we developed a fast and economic procedure based on molecularly imprinted solid-phase extraction (MISPE) for the selective clean-up of betanin and its stereoisomer isobetanin from beetroot extracts. Dipicolinic acid was used as template for the MIP preparation because of its structural similarity with the chromophore group of betanin. The MISPE procedures were fully optimized allowing the almost complete removal of matrix components such as sugars and proteins, resulting in high extraction recovery of betanin/isobetanin in a single step. Moreover, the whole extraction procedure was performed in environmentally friendly solvents with either ethanol or water. Our MISPE method is very promising for the future development of well-formulated beetroot extract with specified betanin/isobetanin content, ready for food or medicinal use

Multiplexed functionalization of nanoelectromechanical systems with photopatterned molecularly imprinted polymers

International audienceImplementing dedicated and reliable biochemical recognition functionalities onto nanoelectromechanical systems (NEMS) is of primary importance for their development as ultra-sensitive and highly-integrated biosensing devices. In this paper, we demonstrate the large-scale and multiplexed integration of molecularly imprinted polymers (MIPs) as highly stable biomimetic receptors onto arrays of nanocantilevers. Integration is carried out by spin-coating and photopatterning the polymer layers before releasing the nanostructures. We demonstrate that these biomimetic layers are robust enough to withstand the wet-etch of the sacrificial layer making this functionalization strategy compatible with further MEMS/NEMS processing. As a proof of concept, we fabricate NEMS resonators coated with a MIP using Boc-L-phenylalanine as the template molecule. We demonstrate the preserved molecular recognition ability of the patterned sensitive layer through the fluorescence detection of dansyl-L-phenylalanine, a fluorescent derivative of the template, and the mechanical integrity of the resonators by means of resonant frequency measurements

Light-triggered switchable graphene-polymer hybrid bioelectronics.

International audienceA light-switchable graphene interface to control and regulate electrobiocatalysis in a nanoconfined space is reported for the first time. The development of switchable and/or tunable interfaces on 2D nanosurfaces endowed with desirable functionalities, and incorporation of these interfaces into remote controlled biodevices, is a rapidly emerging area in bioelectronics

Rapid prototyping of chemical microsensors based on molecularly imprinted polymers synthesized by two-photon stereolithography.

International audienceTwo-photon stereolithography is used for rapid prototyping of submicrometre molecularly imprinted polymer-based 3D structures. The structures are evaluated as chemical sensing elements and their specific recognition properties for target molecules are confirmed. The 3D design capability is exploited and highlighted through the fabrication of an all-organic molecularly imprinted polymeric microelectromechanical sensor