Enzymatic (2R,4R)-Pentanediol Synthesis – “Putting a Bottle on the Table”

(2R,4R)-Pentanediol is an interesting precursor for the synthesis of chiral ligands. A ketoreductase (KRED) was employed for the asymmetric reduction of acetylacetone to this diol. Biocatalysis often suffers from low concentrations of hydrophobic substrates and low stability of the enzyme in unconventional media. Here, we present an engineered KRED variant applicable in a neat substrate system, including upscaling to the multi-liter scale and downstream processing (DSP). Our engineered KRED applied in a neat substrate system is a powerful technique for the synthesis of chiral diols yielding product concentrations of 208 g L⁻¹.     

A new concept for glycosyltransferase inhibitors: nonionic mimics of the nucleotide donor of the human blood group B galactosyltransferase

Glycosyltransferases play an important role in the formation of oligosaccharides and glycoconjugates. To find suitable and selective inhibitors for this class of enzymes is still challenging. Here, we describe a novel concept that allows the design of inhibitors based on the structure of the donor substrate binding pocket. As a first step we describe the design, synthesis and analysis of inhibitors of the human blood group B galactosyltransferase (GTB). This enzyme served as a model system to study the concept, which can be used for easy access of glycosyltransferase inhibitors in general. In silico docking of bicyclic heteroaromatic ligands to GTB and experimental verification of binding affinities by saturation transfer difference NMR (STD NMR) spectroscopy gave 9-N-pentityl uric acid derivatives as non-ionic mimics of UDP. Two derivatives were synthesized and showed inhibitory activity for GTB as determined by competitive STD NMR experiments and by a radiolabeled enzyme assay.     

Compliant membranes improve resolution in full-wafer micro/nanostencil lithography

This work reports on a considerable resolution improvement of micro/nanostencil lithography when applied on full-wafer scale by using compliant membranes to reduce gap-induced pattern blurring. Silicon nitride (SiN) membranes are mechanically decoupled from a rigid silicon (Si) frame by means of four compliant, protruding cantilevers. When pressing the stencil into contact with a surface to be patterned, the membranes thus adapt to the surface independently and reduce the gap between the membrane and the substrate even over large, uneven surfaces. Finite element modeling (FEM) simulations show that compliant membranes can deflect vertically 40 μm which is a typical maximal non-planarity observed in standard Si wafers, due to polishing. Microapertures in the stencil membrane are defined by UV lithography and nanoapertures, down to 200 nm in diameter, using focused ion beam (FIB). A thin aluminium (Al) layer is deposited through both compliant and non-compliant membranes on a Si wafer, for comparison. The blurring in the case of compliant membranes is up to 95% reduced on full-wafer scale compared to standard (non-compliant) membranes.     

Katalytische Dechlorierung von Chlorkohlenwasserstoffen aus kontaminierten Grundw?ssern

Im Rahmen des Forschungsverbundes SAFIRA wurden zwei neue Katalysatorsysteme zur reduktiven Dehalogenierung von CKW mit H2 entwickelt und am Standort Bitterfeld in Grundwasser-Durchflussreaktoren untersucht. Die katalytisch aktive Komponente Pd wurde zum Schutz vor Vergiftung durch ionische Verbindungen zum einen in hydrophobe Polymermembranen eingebettet, zum anderen wurden Pd-Cluster in den Poren hydrophober Zeolithe erzeugt. Die erreichten Standzeiten beider Katalysatoren waren trotzdem bedingt durch die Bildung von H ₂ S und organischen Schwefelverbindungen durch mikrobiologische Sulfatreduktion gering. Die Katalysatoren waren zwar durch eine Spülung mit Hypochlorit regenerierbar, unterlagen aber nach wenigen Tagen erneuter Vergiftung. Der Einsatz von Pd-Katalysatoren unter Bitterfelder Grundwasserbedingungen wird deshalb als problematisch eingesch?tzt.     

Models and computational strategies linking physiological response to molecular networks from large-scale data

An important area of research in systems biology involves the analysis and integration of genome-wide functional datasets. In this context, a major goal is the identification of a putative molecular network controlling physiological response from experimental data. With very fragmentary mechanistic information, this is a challenging task. A number of methods have been developed, each one with the potential to address an aspect of the problem. Here, we review some of the most widely used methodologies and report new results in support of the usefulness of modularization and other modelling techniques in identifying components of the molecular networks that are predictive of physiological response. We also discuss how system identification in biology could be approached, using a combination of methodologies that aim to reconstruct the relationship between molecular pathways and physiology at different levels of the organizational complexity of the molecular network.     

Studies addressing the importance of charge in the binding of fosmidomycin-like molecules to deoxyxylulosephosphate reductoisomerase

Fosmidomycin and its homologue FR900098 are inhibitors of 1-deoxy-D-xylulose-5-phosphate reductoisomerase, which is part of the mevalonate-independent isoprenoid biosynthetic pathway. Replacement of the phosphonate moiety by uncharged sulfone or sulfonamide partial structures resulted in complete loss of activity. Dropping one of the two negative charges resulted in a marked decrease in activity. Through occupation of a hydrophobic binding site, some activity could be regained, leading to compounds with micromolar activity against cultured malaria parasites.