Photoradical-Mediated Catalyst-Independent Protein Cross-Link with Unusual Fluorescence Properties

Photo-actively modified natural amino acids have served as lucrative probes for precise mapping of the dynamics, interaction networks, and turnover of cytosolic proteins both in vivo and ex vivo. In our attempts to extend the utility of photoreactive reporters to map the molecular characteristics of vital membrane proteins, we carried out site-selective incorporation of 7-fluoro-indole in the human mitochondrial outer membrane protein VDAC2 (voltage-dependent anion channel isoform 2), with the aim of generating Trp−Phe/Tyr cross-links. Prolonged irradiation at 282 nm provided us with a surprisingly unusual fluorophore that displayed sizably red-shifted excitation (λ_ex-max=280 nm→360 nm) and emission (λ_em-max=330 nm→430 nm) spectra that was reversible with organic solvents. By measuring the kinetics of the photo-activated cross-linking with a library of hVDAC2 variants, we demonstrate that formation of this unusual fluorophore is kinetically retarded, independent of tryptophan, and is site-specific. Using other membrane (Tom40 and Sam50) and cytosolic (MscR and DNA Pol I) proteins, we additionally show that formation of this fluorophore is protein-independent. Our findings reveal the photoradical-mediated accumulation of reversible tyrosine cross-links, with unusual fluorescent properties. Our findings have immediate applications in protein biochemistry and UV-mediated protein aggregation and cellular damage, opening avenues for formulating therapeutics that prolong cell viability in humans.     

Selective oxidation of HMF to DFF using Ru/γ-alumina catalyst in moderate boiling solvents toward industrial production

Selective oxidation of 5-hydroxymethyl-2-furfural (HMF) to 2,5-diformylfuran (DFF) toward industrial production was studied over Ru supported γ-alumina catalyst using molecular oxygen as an oxidant. From the solvents screening, considering recyclability after reaction, toluene was found to be the best solvent and gave maximum conversion of 99% with 97% DFF selectivity at 130 °C and 40 psi O₂ pressure. Catalyst was washed with NaOH solution of pH = 12 to remove the adsorbed polymer impurities and then reused up to 5 cycles. The product could be purified by simple evaporation of the solvent, which could add advantage for industrial process.     

The influence of annealing on the crystallization and tribological behavior of MWNT/PEEK nanocomposites

In this study, annealing influence on crystallization and scratch behavior of neat and multi‐wall carbon nanotube (MWNT) reinforced poly(ether ether ketone) (PEEK) nanocomposites have been investigated. Crystallization behavior of normal and annealed samples was investigated by using differential scanning calorimeter (DSC). Scratch behavior of normal and annealed samples was investigated by using micro scratch tester. In DSC analysis, it was detected that, melting enthalpy of annealed neat PEEK was increased sharply when compared to neat PEEK. Melting enthalpies of annealed PEEK nanocomposites prepared with addition of up to 1 wt% MWNT were increased with a decreased trend. However, nanocomposites with higher contents of MWNTs (>1 wt%) were dramatically affected by annealing process and melting enthalpy decreased sharply. Friction coefficient values of “annealed MWNT reinforced PEEK composites” were found to be lower than “normal PEEK composites.” Annealing process affects scratch hardness of both annealed and MWNT reinforced PEEK. Annealed nanocomposites with various MWNT concentrations showed higher scratch hardness values than normal PEEK nanocomposites. POLYM. COMPOS., 2011. © 2011 Society of Plastics Engineers     

Fabrication of polypyrrole/graphene oxide nanocomposites by liquid/liquid interfacial polymerization and evaluation of their optical,electrical and electrochemical properties

A novel route has been developed to synthesize polypyrrole (PPy)/graphene oxide (GO) nanocomposites via liquid/liquid interfacial polymerization where GO and initiator was dispersed in the aquous phase and the monomer was dissolved in the organic phase. The synthesized samples were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), ultraviolet–visible absorption (UV–vis), X-ray diffraction (XRD), electrochemical and electrical conductivity measurements. A good dispersion of the GO sheets within the PPy matrix was observed from the morphological analysis. The composites exhibited noticeable improvement in thermal stability and electrical conductivity in comparison to pure polypyrrole. The composites showed excellent electrochemical reversibility at the scan rate of 0.1 V/s and good cyclic stability even up to 100th cycle. Newly developed graphene oxide based polypyrrole composite could be applied in electrochemical energy storage device.     

Hydrogen‐Bonding‐Driven Enantioselective Resolution against the Kazlauskas Rule To Afford γ‐Amino Alcohols by Candida rugosa Lipase

Most lipases resolve secondary alcohols in accordance with the “Kazlauskas rule” to give the R enantiomers. In a similar manner to other lipases, Candida rugosa lipase (CRL) exhibits R enantioselectivity towards heptan‐2‐ol, although the enantiomeric ratio (E) is low (E=1.6). However, unexpected enantioselectivity (i.e., S enantioselectivity, E=58) of CRL towards 4‐(tert‐butoxycarbonylamino)butan‐2‐ol, which has a similar chain length to heptan‐2‐ol, has been observed. To develop a deeper understanding of the molecular basis for this unusual enantioselectivity, we have conducted a series of molecular modeling and substrate engineering experiments. The results of these computational and experimental analyses indicated that a hydrogen bond between the Ser450 residue and the nitrogen atom of the carbamate group is critical to stabilize the transition state of the S enantiomer.     

Phase Transformation of Metastable ZnSnO3 Upon Thermal Decomposition by In‐Situ Temperature‐Dependent Raman Spectroscopy

Temperature‐dependent in‐situ Raman spectroscopy is used to investigate the phase transformation of zinc metastannate (ZnSnO₃) to zinc orthostannate (Zn₂SnO₄) induced upon annealing in the ambient. ZnSnO₃ microcubes (MCs) were synthesized at room temperature using a simple aqueous synthesis process, followed by characterization using electron microscopy, X‐ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and thermogravimetric analysis (TGA). Annealing of the ZnSnO₃ MCs was carried out up to 1000°C, while recording the Raman spectra in‐situ at regular intervals. Phase transformation from metastannate to orthostannate was found to begin around 500°C with an activation energy of ~0.965 eV followed by the recrystallization into the inverse spinel orthostannate phase at ~750°C. Results from this study provide detailed understanding of the phase transformation behavior of perovskite ZnSnO₃ to inverse spinel Zn₂SnO₄ upon thermal annealing.     

Synthesis of opaque and colored hollow polymer pigments

A parametric study was carried out to investigate the effects of process parameters on the properties of opaque and/or colored polymer pigment. The core was synthesized using methyl methacrylate (MMA), methacrylic acid (MAA), and ethylene glycol dimethacrylate, and the shell from styrene. The surfactant/water (S/W) ratio was changed between 0.56 and 1.46, and the best ratio was determined to be 0.97. The increase in the amount of MAA and the pH of the medium increases the diameter of the swollen particles. The hollows could form only when the fraction of MAA in the monomer mixture was above 10%, and pH ≥ 10. The opaque pigment with the largest diameter could be obtained at the S/W ratio of 0.97 and the MAA content of 20%, as 350 nm. It gave an opacity value of 93.8% when mixed with an acrylic resin at 50% by volume. The colored opaque polymer pigments were synthesized by adding copper phthalocyanine into the monomer mixture. The addition of 2% copper phthalocyanine yielded a green‐blue color but 3% yielded blue color. The latter yielded a change in the lightness by a value of ?31.49, and a total color difference of 30.05 compared with that of white opaque polymer pigment. POLYM. ENG. SCI., 57:913–920, 2017. © 2016 Society of Plastics Engineers     

Nonporous monosize polymeric sorbents: Dye and metal chelate affinity separation of lysozyme

Lysozyme adsorption onto dye‐attached nonporous monosize poly(2‐hydroxyethyl‐methacrylate‐methylmethacrylate) [poly(HEMA‐MMA)] microspheres was investigated. Poly(HEMA‐MMA) microspheres were prepared by dispersion polymerization. The monochloro‐triazine dye, Cibacron Blue F3GA, was immobilized covalently as dye–ligand. These dye‐affinity microspheres were used in the lysozyme adsorption–desorption studies. The effect of initial concentration of lysozyme and medium pH on the adsorption efficiency of dye‐attached and metal‐chelated microspheres were studied in a batch reactor. Effect of Cu(II) chelation on lysozyme adsorption was also studied. The nonspecific adsorption of lysozyme on the poly(HEMA‐MMA) microspheres was 3.6 mg/g. Cibacron Blue F3GA attachment significantly increased the lysozyme adsorption up to 247.8 mg/g. Lysozyme adsorption capacity of the Cu(II) incorporated microspheres (318.9 mg/g) was greater than that of the Cibacron Blue F3GA‐attached microspheres. Significant amount of the adsorbed lysozyme (up to 97%) was desorbed in 1 h in the desorption medium containing 1.0M NaSCN at pH 8.0 and 25 mM EDTA at pH 4.9. In order to examine the effects of separation conditions on possible conformational changes of lysozyme structure, fluorescence spectrophotometry was employed. We conclude that dye‐ and metal‐chelate affinity chromatography with poly(HEMA‐MMA) microspheres can be applied for lysozyme separation without causing any significant changes and denaturation. Repeated adsorption/desorption processes showed that these novel dye‐attached monosize microspheres are suitable for lysozyme adsorption. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 76: 115–124, 2000     

Effect of fiber orientation on viscoelastic properties of polymer matrix composites subjected to thermal cycles

Fibers in polymer composites can be designed in various orientations for their usage in service life. Various fiber orientated polymer composites, which are used in aeroplane and aerospace applications, are frequently subjected to thermal cycles because of the changes in body temperatures at a range of −60 to 150°C during flights. It is an important subject to investigate the visco‐elastic properties of the thermal cycled polymer composite materials which have various fiber orientations during service life. Continuous fiber reinforced composites with a various fiber orientations are subjected to 1,000 thermal cycles between the temperatures of 0 and 100°C. Dynamic mechanic thermal analysis (DMTA) experiments are carried out by TA Q800 type equipment. The changes in glass transition temperature (T_g), storage modulus (E′), loss modulus (E′′) and loss factor (tan δ) are inspected as a function of thermal cycles for different fiber orientations. It was observed that thermal and dynamic mechanical properties of the polymer composites were remarkably changed by thermal cycles. It was also determined that the composites with [45°/−45°]_s fiber orientation presented the lowest dynamic mechanical properties. POLYM. COMPOS., 2010. © 2009 Society of Plastics Engineers