Whole-cell Gluconobacter oxydans biosensor for 2-phenylethanol biooxidation monitoring

A microbial biosensor for 2-phenylethanol (2-PE) based on the bacteria Gluconobacter oxydans was developed and applied in monitoring of a biotechnological process. The cells of G. oxydans were immobilized within a disposable polyelectrolyte complex gel membrane consisting of sodium alginate, cellulose sulphate and poly(methylene-co-guanidine) attached onto a miniaturized Clark oxygen electrode, forming whole cell amperometric biosensor. Measured changes in oxygen concentration were proportional to changes in 2-PE concentration. The biosensor sensitivity was 864 nA mM⁻¹ (RSD = 6%), a detection limit of 1 μM, and the biosensor response towards 2-PE was linear in the range 0.02–0.70 mM. The biosensor preserved 93% of its initial sensitivity after 7 h of continuous operation and exhibited excellent storage stability with loss of only 6% of initial sensitivity within two months, when stored at 4 °C. The developed system was designed and successfully used for an off-line monitoring of whole course of 2-PE biooxidation process producing phenylacetic acid (PA) as industrially valuable aromatic compound. The biosensor measurement did not require the use of hazardous organic solvent. The biosensor response to 2-PE was not affected by interferences from PA and phenylacetaldehyde at concentrations present in real samples during the biotransformation and the results were in a very good agreement with those obtained via gas chromatography.     

Novel and Stereospecific Synthesis of (2S)‐3‐(2,4,5‐Trifluorophenyl)propane‐1,2‐diol from D‐Mannitol

A stereospecific synthesis of (2S)‐3‐(2,4,5‐trifluorophenyl)propane‐1,2‐diol from D ‐mannitol has been developed. The reaction of 2,3‐O‐isopropylidene‐D ‐glyceraldehyde with 2,4,5‐trifluorophenylmagnesium bromide gave [(4R)‐2,2‐dimethyl‐1,3‐dioxolan‐4‐yl](2,4,5‐trifluorophenyl)methanol in 65% yield as a mixture of diastereoisomers (1 : 1). The Ph₃P catalyzed reaction of the latter with C₂Cl₆ followed by reduction with Pd/C‐catalyzed hydrogenation gave (2S)‐3‐(2,4,5‐trifluorophenyl)propane‐1,2‐diol with >99% ee and 65% yield.     

Label-free immunosensor for monitoring vitellogenin as a biomarker for exogenous oestrogen compounds in amphibian species

A label-free, optical waveguide lightmode spectroscopy based immunosensor was developed for frog (Bombina orientalis) vitellogenin (Vtg) determination in biological samples as a biomarker for exogenous oestrogen compounds. Antibody against Vtg was produced in rabbits immunised with purified lipovitellin (Lpv), a precursor of Vtg, from the homogenised ovary of oriental fire-bellied toads (B. orientalis). The purified protein and Lpv/Vtg-specific serum were applied in both competitive and direct immunoassay formats using optical waveguide lightmode spectroscopy immunosensor. When measuring Vtg in direct manner, the Lpv antibody (1.76 µg mL^?1) was immobilised on the sensor surface, and the linear measuring range for Vtg was 0.1–10 µg mL^?1. During the competitive measurement, 100 ng mL^?1 Lpv was applied for the immobilisation. The linear measuring range for Vtg was 0.5–50 ng mL^?1. We studied the relative substrate specificity of the antibody, and it was concluded that the method is suitable for the sensitive and selective determination of Vtg levels in toads. Heart, liver and gonad samples from male animals were spiked with Vtg and were analysed using the newly developed method, and female toads and spawn samples were tested and compared to the calibration curve obtained by the spiked samples.     

Determination of acid volatile sulphur in river sediments by ICP-OES coupled with purge and trap method

The developed and tested method is based on the acid volatile sulphur (AVS) releasing from a sample of sediment by acidification with hydrochloric acid and sequential absorption of released hydrogen sulphide into the absorption solution of sodium hydroxide. Total sulphur absorbed in the absorption solution is subsequently determined by inductively coupled plasma optical emission spectrometry (ICP-OES) technique. The method was tested using both model and real samples of the river sediment. Obtained results were compared with the results of iodometric determination as the method routinely used for the AVS determination. In case of using ICP-OES with purge and trap, there is no spectral interference of calcium; therefore for the determination of sulphur, it is possible to use atomic emission line of sulphur S I 180.7 nm. The tested method provides the results comparable with the iodometric method within the range of tested concentrations. The detection limit (LOD) of this method is 0.09 mg· S^2? ? L^?1 in the absorption solution which corresponds to 0.23 mg S^2?·kg^?1 in a sediment. Value of LOD is comparable with the iodometric determination.     

Structural and Functional Implications of the Interaction between Macrolide Antibiotics and Bile Acids

Macrolide antibiotics, such as azithromycin and erythromycin, are in widespread use for the treatment of bacterial infections. Macrolides are taken up and excreted mainly by bile. Additionally, they have been implicated in biliary system diseases and to modify the excretion of other drugs through bile. Despite mounting evidence for the interplay between macrolide antibiotics and bile acids, the molecular details of this interaction remain unknown. Herein, we show by NMR measurements that macrolides directly bind to bile acid micelles. The topology of this interaction has been determined by solvent paramagnetic relaxation enhancements (solvent PREs). The macrolides were found to be bound close to the surface of the micelle. Increasing hydrophobicity of both the macrolide and the bile acid strengthen this interaction. Both bile acid and macrolide molecules show similar solvent PREs across their whole structures, indicating that there are no preferred orientations of them in the bile micelle aggregates. The binding to bile aggregates does not impede macrolide antibiotics from targeting bacteria. In fact, the toxicity of azithromycin towards enterotoxic E. coli (ETEC) is even slightly increased in the presence of bile, as was shown by effective concentration (EC₅₀) values.     

Synthesis of Nature‐Inspired Medium‐Sized Fused Heterocycles from Amino Acids

Herein, we describe the synthesis of molecular scaffolds consisting of medium‐sized fused heterocycles using amino acids, which are some of the most useful building blocks used by nature as well as chemists to create structural diversity. The acyclic precursors were assembled by using traditional Merrifield solid‐phase peptide synthesis, and cyclization was carried out through acid‐mediated tandem endocyclic N‐acyliminium ion formation, followed by nucleophilic addition with internal nucleophiles. The synthesis of molecular scaffolds consisting of seven‐, eight‐, and nine‐membered rings proceeded with full stereocontrol of the newly generated stereogenic center in most cases.     

Transition‐Metal‐Free N‐Arylation of Pyrazoles with Diaryliodonium Salts

A new synthetic method was developed for the N‐arylation of pyrazoles using diaryliodonium salts. The transformation does not require any transition‐metal catalyst and provides the desired N‐arylpyrazoles rapidly under mild reaction condition in the presence of aqueous ammonia solution as a mild base without the use of inert atmosphere. The chemoselectivity of unsymmetric diaryliodonium salts was also explored with large number of examples.     

Swelling behaviors of poly(dodecyl methacrylate-co-methyl eugenol) and poly(dodecyl methacrylate-co-methyl chavicol) gels in essential oil components

Poly(dodecylmethacrylate-co-methyleugenol) P(DDMA-co-Meu) and poly(dodecylmethacrylate-co-methylchavicol) P(DDMA-co-Mch) gels were synthesized in ethanol using free radical cross-linking polymerization method at 60 °C for 24 h in the presence of azobisisobutyronitrile (AIBN) and triethylene glycol dimethacrylate (TEGDMA) as initiator and cross-linker, respectively. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) were used to characterize the gels. These gels were used to investigate swelling behaviors in linalool and methyl eugenol, in essential oil mixture of phenyl propanoid and terpenoid, and also in various solvents. While the equilibrium swelling values (ESV) of both gels were higher in linalool than in methyl eugenol, the result was vice versa in the case of essential oil mixture. ESVs of both gels were also obtained in various solvents with different functional groups and the highest ESV of both gels were obtained in toluene, and the lowest ESV of P(DDMA-co-Meu) and P(DDMA-co-Mch) was in methanol and in ethylene glycol, respectively. While P(DDMA-co-Meu) is sensitive to carbon number change in alcohol, P(DDMA-co-Mch) is less sensitive. In the case of esters, increase of carbon number in functional group causes a more significant change in ESV than an increase of carbon number in aliphatic chain. Experimental results were correlated by the first-order and second-order models. The second-order model was more suitable than the other. While P(DDMA-co-Meu) gel swelled in linalool exhibits a Fickian diffusion character, the diffusion mechanism of the gel in methyl eugenol is a non-Fickian one. In the case of P(DDMA-co-Mch), the result is vice versa.     

Synthesis,characterization, crystal structure,and antimicrobial studies of 2-morpholinoethyl-substituted benzimidazolium salts and their silver(I)-N-heterocyclic carbene complexes

Research on Chemical Intermediates - This article describes synthesis of N-morpholinoethylbenzimidazole (1), 2-morpholinoethyl-substituted benzimidazolium salts (NHC precursors, 2a–c), and...