Crossing the Border: From Keto- to Imine Reduction in Short-Chain Dehydrogenases/Reductases

The family of NAD(P)H-dependent short-chain dehydrogenases/reductases (SDRs) comprises numerous biocatalysts capable of C=O or C=C reduction. The highly homologous noroxomaritidine reductase (NR) from Narcissus sp. aff. pseudonarcissus and Zt_SDR from Zephyranthes treatiae, however, are SDRs with an extended imine substrate scope. Comparison with a similar SDR from Asparagus officinalis (Ao_SDR) exhibiting keto-reducing activity, yet negligible imine-reducing capability, and mining the Short-Chain Dehydrogenase/Reductase Engineering Database indicated that NR and Zt_SDR possess a unique active-site composition among SDRs. Adapting the active site of Ao_SDR accordingly improved its imine-reducing capability. By applying the same strategy, an unrelated SDR from Methylobacterium sp. 77 (M77_SDR) with distinct keto-reducing activity was engineered into a promiscuous enzyme with imine-reducing activity, thereby confirming that the ability to reduce imines can be rationally introduced into members of the “classical” SDR enzyme family. Thus, members of the SDR family could be a promising starting point for protein approaches to generate new imine-reducing enzymes.     

A practical Equivalent Electrical Circuit model for Proton Exchange Membrane Fuel Cell (PEMFC) systems

An alternative Equivalent Electrical Circuit for Proton Exchange Membrane Fuel Cells is modelled in this study. Both I–V characteristics and H₂ consumptions corresponding to generated power under load and no-load conditions are investigated. For this purpose, H₂ consumptions and I–V characteristics of three different sized PEMFCs are tested. There is a very good harmony between the model results and measured values (relative error %0.7, %6.4 and %2.5 for FC-A, FC-B and FC-C respectively). In the proposed model current passes only on parallel resistance and not on serial resistance at no-load condition. Thus, a FC with higher parallel resistance should be preferred. Another key output of this study is that based on the proposed model, performance comparison of FCs can be performed with the parameters defined in this work. Proposals made in this study can easily be used for performance analysis of FCs under for both steady state and transient analysis.     

Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites

Chorismate and isochorismate constitute branch-point intermediates in the biosynthesis of many aromatic metabolites in microorganisms and plants. To obtain unnatural compounds, we modified the route to menaquinone in Escherichia coli. We propose a model for the binding of isochorismate to the active site of MenD ((1R,2S, 5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxycyclohex-3-ene-1-carboxylate (SEPHCHC) synthase) that explains the outcome of the native reaction with α-ketoglutarate. We have rationally designed variants of MenD for the conversion of several isochorismate analogues. The double-variant Asn117Arg–Leu478Thr preferentially converts (5S,6S)-5,6-dihydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHD), the hydrolysis product of isochorismate, with a >70-fold higher ratio than that for the wild type. The single-variant Arg107Ile uses (5S,6S)-6-amino-5-hydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHA) as substrate with >6-fold conversion compared to wild-type MenD. The novel compounds have been made accessible in vivo (up to 5.3 g L⁻¹). Unexpectedly, as the identified residues such as Arg107 are highly conserved (>94 %), some of the designed variations can be found in wild-type SEPHCHC synthases from other bacteria (Arg107Lys, 0.3 %). This raises the question for the possible natural occurrence of as yet unexplored branches of the shikimate pathway.     

Effect of Boronizing on Operating Stability of Steel AISI 304L under Erosion Impact of Hard Particles

Metal Science and Heat Treatment - The effect of solid boronizing at 950°C for 2 and 4 h on the phase composition, microstructure, hardness and abrasive wear of steel AISI 304L is studied...     

Flexible poly(styrene-ethylene-butadiene-styrene) hybrid nanofibers for bioengineering and water filtration applications

Among the thermoplastic elastomers that play important roles in the polymer industry due to their superior properties, styrene-based species and polyurethane block copolymers are of great interest. Poly(styrene-ethylene-butadiene-styrene) (SEBS) as a triblock copolymer seems to have the potential to meet many demands in different applications due to various industrial requirements where durability, biocompatibility, breaking elongation, and interfacial adhesion are important. In this study, the SEBS triblock copolymer was functionalized with natural (Satureja hortensis, SH) and synthetic (nanopowder, TiO₂) agents to obtain composite nanofibers by electrospinning and electrospraying methods for use in biomedical and water filtration applications. The results were compared with thermoplastic polyurethane (TPU) composite nanofibers, which are commonly used in these fields. Here, functionalized SEBS nanofibers exhibited antibacterial effect while at the same time improving cell viability. In addition, because of successful water filtration by using the SEBS composite nanofibers, the material may have a good potential to be used comparably to TPU for the application.     

A Potent Mimetic of the Siglec-8 Ligand 6’-Sulfo-Sialyl Lewisx

Siglecs are members of the immunoglobulin gene family containing sialic acid binding N-terminal domains. Among them, Siglec-8 is expressed on various cell types of the immune system such as eosinophils, mast cells and weakly on basophils. Cross-linking of Siglec-8 with monoclonal antibodies triggers apoptosis in eosinophils and inhibits degranulation of mast cells, making Siglec-8 a promising target for the treatment of eosinophil- and mast cell-associated diseases such as asthma. The tetrasaccharide 6’-sulfo-sialyl Lewis^x has been identified as a specific Siglec-8 ligand in glycan array screening. Here, we describe an extended study enlightening the pharmacophores of 6’-sulfo-sialyl Lewis^x and the successful development of a high-affinity mimetic. Retaining the neuraminic acid core, the introduction of a carbocyclic mimetic of the Gal moiety and a sulfonamide substituent in the 9-position gave a 20-fold improved binding affinity. Finally, the residence time, which usually is the Achilles tendon of carbohydrate/lectin interactions, could be improved.