Lewis Acid‐Catalyzed or Base‐Promoted Regioselective Cycloisomerization of N‐Imidoyl‐o‐alkynylanilines for Synthesis of N‐Imidoyl‐(1 H)‐indoles and 4‐Alkylidene‐3,4‐dihydroquinazolines

Product selectivity control for the synthesis of imidoylindoles and 4‐alkylidenedihydroquinazolines from N‐imidoyl‐o‐alkynylanilines via silver triflate‐catalyzed cycloisomerization or tetrabutylammonium fluoride‐promoted cyclization is described. The product selectivity depends mainly on the catalyst/promoter used, and on the substituents on the alkyne and amidine functions of the substrates.

     

Polymerization of β-lactoglobulin and bovine serum albumin at oil-water interfaces in emulsions by transglutaminase

Polymerization of β-lactoglobulin and bovine serum albumin at the oil—water interfaces in n-tetradecane-in-water emulsions induced by the transglutaminase reaction was studied. The emulsions were incubated with transglutaminase for various times, and adsorbed and unadsorbed protein fractions at the oil—water interfaces were analyzed by sodium dodecyl sulfate—polyacrylamide gel electrophoresis. While only monomers were detected in the unadsorbed fractions, polymers were observed in the adsorbed fractions of the both proteins. The sizes and amounts of the polymers increased with incubation time. The incubation with transglutaminase caused much flocculation of the emulsion stabilized by β-lactoglobulin. An increase in viscosity was also observed with the flocculation. The flocculation was probably initiated by the formation of ε-(γ-glutamyl)-lysyl isopeptide bonds between β-lactoglobulin molecules adsorbed on different oil droplets. In the case of the emulsion stabilized by bovine serum albumin, however, the flocculation and the increase in viscosity occurred to only limited extents by the transglutaminase reaction. This suggests that ε-(γ-glutamyl)-lysyl isopeptide bonds induced by the transglutaminase reaction were formed only between neighboring molecules of bovine serum albumin on the same droplet.     

Amino Acid Absorption in Portal Blood After Duodenal Infusions of a Soy Protein Hydrolysate Prepared by a Novel Soybean Protease D3

ABSTRACT:  The intestinal absorption of amino acids from decapeptide was investigated in rats under unrestrained conditions. The soy protein hydrolysate utilized in the experiment was produced by a novel soybean protease D3. The enzymatic features of protease D3 showed high homology with cathepsin L and cathepsin K and the average molecular weight of D3 hydrolysate is approximately 1200. We compared the intestinal absorption of D3 hydrolysate in portal blood with that of an amino acids mixture and soy protein with the same amino acid composition by determining the concentration of individual amino acids after a single administration of a nitrogen source. The absorptive velocity and intensity of each amino acid were calculated from its rate of elevation in the portal blood. And in most cases, these were higher in the D3 hydrolysate than in amino acids mixture and protein. The proportion of the amount of each amino acid absorbed in portal blood from D3 hydrolysate was much more like the composition of the administrated amino acids than like that from the amino acids mixture. The result of in vitro digestion assay indicated that D3 hydrolysate was hydrolyzed easier than the hydrolysates produced by microbial proteases. This is the first report to demonstrate that the D3 hydrolysate, which contains decapeptide as a dominant fraction, was more rapidly utilized than the amino acids mixture and protein as is the case with di-, tripeptides. This suggested that this hydrolysate could be available for nutraceutical use as well as use in nutritious foods for athletes and patients.     

Characteristic Property of Low Bitterness in Protein Hydrolysates by a Novel Soybean Protease D3

ABSTRACT:  Enzymatic hydrolysis is 1 means of improving the functional properties of food protein; however, in most cases, bitter peptides are generated by such treatment, and the resulting product is therefore not acceptable as a food ingredient. We have already reported a novel cysteine protease, D3, purified from germinating soybean cotyledons. Because of its substrate specificities, most hydrophobic amino acid residues in the hydrolysate are presumed not to be located at the peptide termini. It was therefore expected that protein hydrolysate by protease D3 would taste less bitter than other enzymatic hydrolysates. The objective of this study was to demonstrate the low bitterness of protein hydrolysates by protease D3. For that purpose, soy protein and casein hydrolysates were prepared with treatment of protease D3, subtilisin, pepsin, trypsin, and thermolysin, respectively. The bitterness of these hydrolysates was evaluated by measuring points of subjective equality (PSE). The PSE value demonstrated that the protein hydrolysates by protease D3 were significantly less bitter than the other enzymatic hydrolysates, indicating that the products had a taste mild enough to be acceptable as a less-bitter peptide food ingredient. These results suggested that a prominent feature of protease D3 was its capacity to produce less-bitter peptides. Therefore, it is thought that protease D3 could be applied to produce protein hydrolysates for use as ingredients in a variety of food products.     

Formation of bromo-substituted triclosan during chlorination by chlorine in the presence of trace levels of bromide

The side reactions of triclosan (2,4,4'-trichloro-2'-hydroxydiphenyl ether, TC) and chlorine in the presence of sodium chloride were investigated. In the absence of sodium chloride, three chloro-derivatives of TC, 2',3,4,4'-tetrachloro-2-hydroxydiphenyl ether (3-Cl-TC), 2',4,4',5-tetrachloro-2-hydroxydiphenyl ether (5-Cl-TC), and 2',3,4,4',5-pentachloro-2-hydroxydiphenyl ether (3,5-Cl(2)-TC) were formed, whereas in the presence of sodium chloride, 3-bromo-2',4,4'-trichloro-2-hydroxydiphenyl ether (3-Br-TC), 5-bromo-2',4,4'-trichloro-2-hydroxydiphenyl ether (5-Br-TC), (3 or 5)-bromo-2',4,4',(5 or 3)-chloro-2-hydroxydiphenyl ether ((3,5)-(BrCl)-TC), and 3,5-dibromo-2',4,4'-trichloro-2-hydroxydiphenyl ether (3,5-Br(2)-TC) were additionally formed. Radiochemical neutron activation analysis indicated that 1g of commercially available sodium chloride contained 73 microg of bromide and the bromide ion was determined to be the source of the side reactions. The rate of decrease of TC due to reaction with chlorine was greatly accelerated by the presence of bromide ion in the system: the rate with only 1 x 10(-5) M bromide ion was three times the rate in the absence of bromide.     

Microstructures and Thermoelectric Properties of Sintered Misfit-Layered Cobalt Oxide

Misfit-layered cobalt oxide Ca₃Co₄O₉ is considered to be a prospective material for thermoelectric conversion. The thermoelectric properties are anisotropic owing to its anisotropic crystal structure. The crystal has preferred thermoelectric properties along the a–b plane. Therefore, the thermoelectric properties are improved and controlled by the degree of orientation of the sintered sample. In the present work, Sr-doped misfit cobalt oxide Ca_2.7Sr_0.3Co₄O₉ was prepared by solid-phase reaction, followed by uniaxial compression molding and sintering at 1173 K. The Seebeck coefficient α, electrical resistivity ρ, and dimensionless figure of merit ZT were measured as a function of the compression pressure applied in the uniaxial molding. α, ρ, and ZT as functions of the degree of orientation and the relative density are experimentally clarified and explained by calculations using the compound model.     

The Effective Methods in Refolding and Activation of Cathepsin L-like Soybean Protease D3

A novel cysteine protease D3, which was purified from germinating soybean cotyledons, showed high homology with cathepsin L and cathepsin K. In our previous study, because of the specificity of the enzyme, hydroly‐sates treated with D3 treatment showed a prominent property of less bitterness than other hydrolysates treated with commercially available proteases. However, active recombinant D3 prepared from Escherichia coli inclusion bodies was so intricate and less productive that it made further studies on this protease and hydrolysates difficult. In the concrete, the refolding process of the immature proD3 from inclusion bodies takes more than a day, and autocatalytic activation of refolded immature proD3 at low pH was difficult to control. In this study, we aimed to establish an efficient refolding and activating method of protease D3. In the refolding step, the procedures could be simplified by using a size‐exclusive column‐based method. In the activation step from immature proD3, we utilized another protease, subtilisin, rather than autocatalytic activation by D3 itself. After subtilisin treatment, the peptide having 12 amino acids‐length of N‐terminal pro sequence was initially cleaved, and residual proD3 showed only a half proteolytic activity of active D3. However, when the pH was shifted lower (pH4.5), D3 automatically changed to have the same proteolytic activity as active one, and this activated recombinant had the same N‐terminal sequence as purified D3 from germinating soybean cotyledons. By using this method, all preparation processes of D3 from inclusion bodies to active D3 could be completed within a few hours, and it became possible to carry out the investigation on hydrolysates on a large scale.     

Simultaneously Measuring Lateral and Vertical Forces with Accurate Gap Control for Clarifying Lubrication Phenomena at Nanometer Gap

A novel method for simultaneous measurement of vertical and lateral forces with high sensitivity and accurate gap control is presented. It combines a double-ended tuning fork resonator for vertical force measurement with an optical fiber probe for lateral force measurement. High force sensitivity of several tens of nano Newons with accurate gap control of less than 1 nm was successfully attained. Simultaneous measurement using this novel method revealed that viscoelasticity increased with decrease of the sliding gap. Moreover, the method first revealed that the lubricant pressure generated by the wedge effect can increase due to the confinement at small gaps, even if the lubricant pressure is so low that the viscosity rise due to the pressure is much small. This method is expected to be useful for clarifying the lubrication phenomena at nanometer sliding gaps, which are not yet fully understood due to the difficulty of measurement.     

Dumb-bell shaped tuberculous abscess across the greater sciatic notch compressing both sciatic nerves

We report an instructive case of a 65-year-old man who presented with a dumb-bell shaped tuberculous abscess across the greater sciatic notch bilaterally compressing both sciatic nerves. Clinical symptoms progressed slowly and mimicked lumbar radiculopathy, thus delaying an accurate diagnosis. Anterolateral retroperitoneal and posterolateral gluteal approaches of the greater sciatic notch as well as the acetabulum on both sides were followed in order to provide safe viewing and resection of the abscess. The abscess wall was adherent to the sciatic nerve and surrounding blood vessels. The symptoms completely disappeared after resection of the abscess.