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.     

On the Enzymatic Formation of Metal Base Pairs with Thiolated and pKa-Perturbed Nucleotides

The formation of artificial metal base pairs is an alluring and versatile method for the functionalization of nucleic acids. Access to DNA functionalized with metal base pairs is granted mainly by solid-phase synthesis. An alternative, yet underexplored method, envisions the installation of metal base pairs through the polymerization of modified nucleoside triphosphates. Herein, we have explored the possibility of using thiolated and pK_a-perturbed nucleotides for the enzymatic construction of artificial metal base pairs. The thiolated nucleotides S2C, S6G, and S4T as well as the fluorinated analogue 5FU are readily incorporated opposite a templating S4T nucleotide through the guidance of metal cations. Multiple incorporation of the modified nucleotides along with polymerase bypass of the unnatural base pairs are also possible under certain conditions. The thiolated nucleotides S4T, S4T, S2C, and S6G were also shown to be compatible with the synthesis of modified, high molecular weight single-stranded (ss)DNA products through TdT-mediated tailing reactions. Thus, sulfur-substitution and pK_a perturbation represent alternative strategies for the design of modified nucleotides compatible with the enzymatic construction of metal base pairs.     

Limitations of Preferential Enrichment: A Case Study on Tryptophan Ethyl Ester Hydrochloride

Since eutomer and distomer show different biological effects, access to pure enantiomers is essential. Preferential enrichment is a relatively new process of chiral resolution developed in the 90s. Extension of its scope was attempted with tryptophan ethyl ester hydrochloride. This salt meets one of the most important requirements for preferential enrichment, namely, a larger solubility for the pure enantiomers with reference to the racemic compound. But, due to the absence of a solid solution, this salt cannot perform preferential enrichment. This impossibility was endorsed by semi‐empirical molecular modeling.     

Eumelanin for nature‐inspired UV‐absorption enhancement of plastics

In the human body, the black‐brown biopigment eumelanin blocks harmful ultraviolet (UV) radiation. In the plastics industry, additives are often added to polymers to increase their UV‐absorption properties. We herein report an assessment of the biopigment eumelanin as a nature‐inspired additive for plastics to enhance their UV absorption. Since eumelanin is produced by natural sources and is nontoxic, it is an interesting candidate in the field of sustainable plastic additives. In this work, the eumelanin‐containing films of commercial ethylene–vinyl acetate copolymer, a plastic used for packaging applications, were obtained by melt compounding and compression molding. The biopigment dispersion in the films was improved by means of the melanin free acid treatment. It was observed that eumelanin amounts as low as 0.8 wt% caused an increase of the UV absorption, up to one order of magnitude in the UVA range. We also evaluated the effect of eumelanin on the thermal stability and photostability of the films: the biopigment proved to be double‐edged, working both as UV‐absorption enhancer and photo‐prooxidant, as thermogravimetric analysis and infrared spectroscopy revealed. © 2019 Society of Chemical Industry     

Analysis of Damage on the Streptococcus mutans Immersed in Ozonated Water: Preliminary Study for Application as Mouth Rinse

Ozonated water has been demonstrated to induce significant results in terms of the elimination of microorganisms. The present study assessed the damage to Streptococcus mutans after exposure to ozonated water; the ozone generator was adjusted to provide an outlet concentration of 60 mg/L, the samples were submitted to different ozonation times 1, 2, 4, 6, and 10 mi. Scanning electron microscopy and atomic force images were obtained to identify damage to the bacteria, followed by reactive oxygen species (ROS) evaluation and microbial viability. The results showed a significant reduction in viability and the images evidenced the generation of gaps on the microbial wall and surface layer alterations. Ozone can induce significant damage to S. mutans, thus suggesting that the use of ozonated water to prevent carious lesion formation is extremely promising.     

Volatile Profiles of Rapeseed Oil Flavored with Basil,Oregano, and Thyme as a Function of Flavoring Conditions

The flavoring of oils with herbs gives a specific taste and aroma to the oils and may increase their antimicrobial and antioxidant activity. The volatile aroma compounds in flavored rapeseed oil were studied as a function of flavoring conditions, by means of headspace solid-phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS) analysis. Rapeseed oils were flavored with dried basil, oregano, and thyme at two different concentrations (3 and 6%) and were heated at different temperatures and for various time intervals, followed by filtration. In the headspace of flavored oils, the main volatiles of the dried herbs were detected. In general, the share of monoterpenes in the headspace of flavored oil was higher than in the original dried herbs, while sesquiterpenes and phenolic compounds were detected to a considerably lower extent in the oil than in the herbs. The concentrations of the volatiles detected in the oil increased with increasing heating time and temperature.     

Experimental and modelling of supercritical oil extraction from rapeseeds and sunflower seeds

The supercritical oil extraction from oleaginous seeds (sunflower and rapeseeds) is presented here through experimental and modelling results. The experimental setup allows an accurate following of the mass of the oil extracted and to derive the experimental influences of pressure, temperature and supercritical CO₂ flowrate on the extraction curves. These parameters are very sensitive and highlight the necessity of precise optimisation of experimental conditions. In order to complete the behaviour of supercritical fluids extraction, an improved modelling is proposed. The modelling basic equations are based on others modelling published previously. In this work, the determination of several parameters comes from correlations and the other constants are fitted with all the experimental results. Thus the modelling is more representative and predictive as other ones. The modelling results present a good agreement with the experimental results, and hence it can be used for the dimensioning of some extraction autoclaves.     

Inkjet printing of photocatalytically active TiO2 thin films from water based precursor solutions

In this work, aqueous chemical solution deposition route suited for inkjet printing is used for the synthesis of photocatalytically active TiO₂ coatings. Environmentally friendly precursor solutions with electromagnetic ink-jet printing, allows cheap and simple processing of TiO₂ films on glass. The hydrolysis reaction of water sensitive titanium alkoxide (Ti-alkoxide) precursor is controlled by adding complexing agents as citric acid and triethanolamine prior to water addition, and aqueous stable solutions are achieved. The pH of the solutions is brought to neutral to guarantee flexible processing, avoid damage to substrates and equipment. Solution parameters are adapted to obtain optimal gelation conditions and good jettability. The influence of processing parameters on the phase formation and surface morphology is studied by thermogravimetric analysis and differential thermal analysis (TGA/DTA), X-ray diffraction (XRD), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The photocatalytic activity of the films is evaluated by the degradation of methyl orange.     

Sintering of α-alumina for highly transparent ceramic applications

In this paper we present our results on the sintering of α alumina ceramics by hot isostatic pressing. It describes a simple method for obtaining precise relative density values on our almost 100% dense samples. Then, transparency results are discussed with respect to grain size and residual porosity measurements, comparing them to scattering calculations. Our results are not far from the best reported transmission values: almost 60% for a 1 mm thick sample. The other 40% diffuse light comes from the birefringence of alumina for the most part. However, they are transparent enough to see detailed structures at several kilometers through them.     

The inverse type II β-turn on D-Trp-Phe, a pharmacophoric motif for MOR agonists

Herein we propose the D ‐Trp‐Phe sequence within an inverse type II β‐turn as a new kind of pharmacophoric motif for μ‐opioid receptor (MOR) cyclopeptide agonists. Initially, we observed that c[Tyr‐D ‐Pro‐D ‐Trp‐Phe‐Gly] ( 4 ), an analogue of endomorphin‐1 (H‐Tyr‐Pro‐Trp‐Phe‐NH₂) lacking the crucial protonatable amino group of Tyr 1, is a MOR agonist with 10^?8 M affinity. Molecular docking analysis suggested that the relevant interactions with the receptor involve D ‐Trp‐Phe. The bioactive conformation of this region was investigated by selected derivatives of 4 designed to adopt an inverse type II β‐turn. These efforts led to c[Tyr‐Gly‐D ‐Trp‐Phe‐Gly] ( 14 ) and to the cyclotetrapeptide c[D ‐Asp‐1‐amide‐β‐Ala‐D ‐Trp‐Phe] ( 15 ), showing improved nanomolar affinity. Both 14 and 15 selectively bind MOR, as they have negligible affinity for the κ‐ and δ‐opioid receptors. Both 14 and 15 behave as partial MOR agonists in functional assays. Conformational and docking analyses confirm the role of the inverse β‐turn in binding. These results indicate that the D ‐Trp‐Phe inverse β‐turn structure can be used for designing non‐endomorphin‐like peptidomimetic opioid agonists in general, characterized by an atypical mechanism of interaction between ligand and receptor.