Higher order structure of proteins solubilized in AOT reverse micelles

The higher order structure of proteins solubilized in an bis(2-ethylhexyl) sulfosuccinate sodium (AOT) reverse micellar system was investigated. From circular dichroic (CD) measurement, CD spectra of cytochrome c, which is solubilized at the interface of reverse micelles, markedly changed on going from buffer solution to the reverse micellar solution, and the ellipticity values in the far- and near-UV regions decreased with decreasing the water content (W₀: molar ratio of water to AOT), indicating that the secondary and tertiary structures of cytochrome c changed with the water content. The ellipticity of ribonuclease A, which is solubilized in the center of micellar water pool, in the near-UV region was dependent on W₀ and became minimum when W₀ of ca. 8 while the ellipticity in the far-UV region was almost constant, indicating that the tertiary structure of ribonuclease A was affected by the water content, but the secondary structure was conserved. The degree of curvature of the micellar interface appears to influence the protein structure because the reverse micelle size is linearly proportional to the W₀ value. As evidence of this, when the micelle size was comparable to the protein's dimensions, the structures were more affected by the water content. Judging from the dependence of the factor influencing the protein structure on the protein species, the location of solubilized protein in reverse micelles is significantly related to whether the protein structure in the system is affected by the micellar interface. In the cases of cytochrome c and lysozyme, the ellipticity against W₀ was dependent on the AOT concentration. In contrast, ribonuclease A gave very similar ellipticity values whatever the AOT concentration. In the n-hexane micellar system, cytochrome c exhibited lower ellipticity values and ribonuclease A in the lower W₀ range (W₀ < ca. 8) higher ellipticity values. These results indicated that the interaction between the protein and the micellar interface is a dominant factor influencing the protein structure in reverse micelles, and that it is governed by the location of solubilized proteins and the state of the micellar interface.     

A type of Strassen's theorem for positive vector measures with values in dual spaces

In this paper, we extend a type of Strassen's theorem for the existence of probability measures with given marginals to positive vector measures with values in the dual of a barreled locally convex space which has certain order conditions. In this process of the extension we also give some useful properties for vector measures with values in dual spaces.

     

A new family of glucose-1-phosphate/glucosamine-1-phosphate nucleotidylyltransferase in the biosynthetic pathways for antibiotics

Aminoglycoside antibiotics are composed of aminosugars and a unique aminocyclitol aglycon including 2-deoxystreptamine (DOS), streptidine, actinamine, etc., and nucleotidylyltransferases, sugar modifying enzymes, and glycosyltransferases appear to be essential for their biosynthesis. However, the genes encoding those enzymes were unable to be identified by a standard homology search in the butirosin biosynthetic btr gene cluster, except that the btrM gene appeared to be a glycosyltransfease. Disruption studies of the btrD gene indicated that BtrD was involved in the supply of a glycosyl donor immediately prior to the glycosylation of DOS giving paromamine. As anticipated, BtrD expressed in Escherichia coli was able to catalyze UDP-D-glucosamine formation from D-glucosamine-1-phosphate and UTP. Both dTTP and UTP were good NTP substrates, and D-glucose-1-phosphate and D-glucosamine-1-phosphate were good sugar phosphates for the enzyme reaction. This finding is the first to identify an enzyme which activates a sugar donor in the DOS-containing antibiotics. Interestingly, BtrD homologues have been reported as functionally unknown open reading frames (ORFs) in the biosynthetic gene clusters for several antibiotics including teicoplanin, balhimycin, chloroeremomycin, and mitomycin C. It appears therefore that gene clusters for antibiotic biosynthesis provide their own nucleotidylyltransferases, and the BtrD homologues are among the secondary metabolism specific enzymes.     

Reactivity of palladium nanoparticles supported on a microemulsion-based organogel network in supercritical carbon dioxide

Palladium (Pd) nanoparticles were prepared using the phase transfer method and coated with alkylamines as stabilizing agents stably dispersed in nonpolar solvents. Spherical Pd nanoparticles with an average diameter of 4 nm and a relatively narrow size distribution were obtained using hexylamine or dodecylamine, and they were successfully incorporated in microemulsion-based gelatin organogel (OG); also, an OG network containing Pd nanoparticles was prepared via drying. For the Mizoroki-Heck cross-coupling reaction of iodobenzene with methyl acrylate in supercritical carbon dioxide, the Pd nanoparticles in the OG network exhibited much higher reactivity than those in powder state. Preparation conditions of OG (e.g., gelatin concentration) affected the apparent reactivity of the supported Pd nanoparticles. The Pd nanoparticles in the OG network with high gelatin concentration were recycled with no appreciable change of reactivity. In contrast, the reactivity of the Pd nanoparticles with low gelatin concentration decreased during recycling.     

Rapid measurement of oestradiol and oestriol by high-performance liquid chromatography after automatic pretreatment

A fluorometric liquid chromatographic method was developed for measurements of unconjugated oestradiol and oestriol in the serum of pregnant women. The serum samples are injected directly into the apparatus and pass to a pretreatment column, where oestrogens are adsorbed while hydrophilic components such as proteins and carbohydrates are not. The oestrogens then pass into a separation column containing a new type of polymer gel. The mobile phase consists of an acetonitrile-water mixture, and separation is achieved by a reversed-phase mechanism. The eluent is monitored for fluorescence. Data on the reproducibility and recovery by this method and the correlation of values with those obtained by radioimmunoassay are reported. Results on the increases of oestradiol and oestriol in the serum during pregnancy are also reported.     

Extraction-spectrophotometric determination of traces of antimony in copper and lead metals and in lead-base alloy with pyrocatechol violet and tri-n-octylamine

A sensitive spectrophotometric method is described for the determination of antimony in copper and lead metals and in lead-base alloy. Optimal conditions have been established for the extraction and determination of antimony. Antimony (III) is extracted from a potassium iodide—sulfuric acid or a hydrobromic—sulfuric acid medium with toluene and converted to an antimony-pyrocatechol violet (PV) complex. The complex is then extracted with tri-n-octylamine (TOA) and the absorbance of the resulting ternary Sb(III)—PV-TOA complex is measured at 555 nm. As little as 0.5 p.p.m. of antimony in copper metal and 0.2 p.p.m. of antimony in lead metal and lead-base alloy can be determined.     

Characterization and Identification of Bioactive Polyphenols in the Trapa bispinosa Roxb. Pericarp Extract

In this study, we present the isolation and characterization of the structure of six gallotannins (1–6), three ellagitannins (7–9), a neolignan glucoside (10), and three related polyphenolic compounds (gallic acid, 11 and 12) from Trapa bispinosa Roxb. pericarp extract (TBE). Among the isolates, the structure of compound 10 possessing a previously unclear absolute configuration was unambiguously determined through nuclear magnetic resonance and circular dichroism analyses. The α-glucosidase activity and glycation inhibitory effects of the isolates were evaluated. Decarboxylated rugosin A (8) showed an α-glucosidase inhibitory activity, while hydrolyzable tannins revealed stronger antiglycation activity than that of the positive control. Furthermore, the identification and quantification of the TBE polyphenols were investigated by high-performance liquid chromatography coupled to ultraviolet detection and electrospray ionization mass spectrometry analysis, indicating the predominance of gallic acid, ellagic acid, and galloyl glucoses showing marked antiglycation properties. These findings suggest that there is a potential food industry application of polyphenols in TBE as a functional food with antidiabetic and antiglycation activities.     

Synthesis and Reactivity of 5,10,15‐Triaryl Doubly N‐Confused Bilanes

A series of 5,10,15‐tris(pentafluorophenyl) doubly N‐confused bilanes were synthesized in a stepwise manner with the aid of sterically demanding N‐protecting groups, in which the difference in reactivity between regular pyrrole and N‐confused pyrrole plays a crucial role in the synthetic strategy. Some doubly N‐confused bilanes were converted into porphyrinoids or a unique 2:2 copper(II) complex with a helical structure. In addition, the conformations and electronic states of the doubly N‐confused bilanes were investigated theoretically, giving fruitful information about the effect of confusion on the bilane skeleton.     

Predictive elucidation of conformational characteristics and configurational properties of poly(1-methylphosphirane) and poly(1-phenylphosphirane) as a molecular design

Conformational characteristics and configurational properties of poly(1-methylphosphirane) (PMePP) and poly(1-phenylphosphirane) (PPhPP) have been predictively elucidated by the refined rotational isomeric state scheme coupled with ab initio molecular orbital and density functional calculations. The lone pair of the phosphorus atom adopts an sp hybrid orbital. Owing to the high s character (50%), the polyphosphiranes exhibit low proton (hydrogen) affinities, and hence the lone pair does not form any intramolecular attractive interactions with hydrogen. As the meso-diad probability varies from 0 (syndiotactic) to 1 (isotactic), the characteristic ratio of PMePP slightly increases from 6.7 to 7.4, whereas that of PPhPP considerably decreases from 38 to 7.3. The large dimension of syndiotactic PPhPP is chiefly due to π-π attractions formed between adjacent phenyl groups. The trivalent phosphorus atom may be bonded to heavy, noble, and transition metals but readily or gradually oxidized. The usefulness and necessity of the polyphosphiranes have been assessed.