Relationship between the glass transition of soy protein and molecular structure

The change in molecular structure of the soy protein samples as a result of the microbial transglutaminase treatment was studied using solid-state (13)C NMR spectroscopy and circular dichroism (CD), and the relation to the glass transition temperature (T(g)) was examined. From NMR measurements, the structure of the local region of the C(alpha) methine was observed to change, and the region had relatively high mobility. From CD measurements, the structural change seemed to be caused by the change in the secondary structure (disintegration of the beta-structure). By comparison with the T(g) of another protein, the state of the secondary structure of a protein was suggested to be a key in determining its T(g).     

Relationship between antibacterial activity of (+)-catechin derivatives and their interaction with a model membrane

(+)-Catechin derivatives with different alkyl chain lengths were synthesized from (+)-catechin and various straight chain alkylaldehydes in the presence of methyl mercaptan, and their antibacterial activities against Gram-positive bacteria were evaluated. The antibacterial activity increased markedly with elongation of the alkyl chain lengths of the derivatives and reached a maximum at a chain of four to seven carbons. Subsequently, interaction of the (+)-catechin derivatives with a model membrane using liposome was investigated. The derivatives with a chain of three carbons or more were found to have very strong affinity for the membrane. The injury action of the derivatives against the membrane was examined with liposome in which calcein was enclosed as a fluorescent indicator. The leakage was observed in the derivatives with chain lengths of four carbons or more. Particularly the derivatives with chains longer than five carbons are considered to destroy the liposome membrane judging from the degree of the fluorescent leakage. These results implied that the lipophilicity and disrupting ability of the (+)-catechin derivatives to the liposome membrane participate in their antibacterial activity.     

Determination of tri-n-butyltin and di-n-butyltin compounds in fish by gas chromatography with flame photometric detection

An analytical method is described for the simultaneous quantitative determination of tri-n-butyltin and di-n-butyltin compounds in fish. The sample was extracted with 0.5N HCl-methanol, and the methanol solution was extracted with hexane. The extract was purified by gel permeation chromatography and treated with Grignard reagent to yield the methyl derivatives, which were determined by gas chromatography with flame photometric detection operated in the tin mode (610 nm). Recoveries of tri-n-butyltin chloride (Bu3SnCl) and di-n-butyltin dichloride (Bu2SnCl2) spiked to fish at the levels of 0.2 and 1.0 ppm ranged from 80 to 105%. Detection limits were 0.02 micrograms/g for both compounds. Tri-n-butyltin compounds equivalent to Bu3SnCl levels of 0.07-2.0 ppm and di-n-butyltin compounds equivalent to Bu2SnCl2 levels of 0.02-0.11 ppm were found in reared yellowtails, and these values showed good agreement with the results from gas chromatographic-mass spectrometric analysis.     

Pressure-induced denaturation of monomer beta-lactoglobulin is partially irreversible: comparison of monomer form (highly acidic pH) with dimer form (neutral pH)

This study was conducted to assess the effect of high hydrostatic pressure on monomer beta-lactoglobulin (BLg) at acid pH by fluorescence spectroscopy under pressure and by circular dichroism (CD) and (1)H NMR spectroscopies after release of pressure. The intrinsic (tryptophan) fluorescence measurement and the study of 8-anilinonaphthalene-1-sulfonate (ANS) binding to BLg indicated that at pH 2.0 the recovery of center of spectral mass or ANS fluorescence was almost complete upon pressure release. No difference in (1)H NMR spectra was observed between pressurized and unpressurized BLg. In addition, NMR detection of the H/D exchange of aromatic protein indicated that the conformation of the vicinity of tryptophan residues could be refolded almost completely after release of pressure. These results seemingly confirm that the pressure-induced denaturation of BLg at pH 2.0 is reversible. However, cis-parinaric acid binding ability of pressurized BLg was largely lost, although its retinol binding ability was the same as its unpressurized one. Furthermore, CD spectra of the far-UV region and 2D NMR spectra evidently revealed the difference in the conformation of the molecule between unpressurized and pressurized BLg. These results are interpreted as an existence of partially fragile structure in the BLg molecule by high pressure.     

Nitrogen Retention in Japanese Cedar Stands in Northern Honshu, with High Nitrogen Deposition

High nitrogen, especially ammonium, input has been observed in Schichinohe, Aomori Prefecture, northeastern Japan. A monitoring study on precipitation, throughfall, and stream water has been carried out to estimate the stage of nitrogen saturation since 1996. Fifty-two to 70% of nitrogen input in throughfall was retained in forest ecosystems. Nitrate concentration in stream water tended to decrease throughout the study. There was no symptom of nitrogen saturation at Japanese cedar stands in Shichinohe, although high nitrogen input in open bulk has been observed. Ammonium (NH₄ ⁺) was retained in the canopy. The ratio of NH₄ ⁺ input in throughfall to that by open bulk was 0.40 – 0.47. Total inorganic nitrogen input under the canopy amounted 0.68 – 0.72 kmolc ha^?1 yr^?1 (9.6 – 10.0 kg N ha^?1 yr^?1). Our results suggests that atmospheric nitrogen input has benefitted the three growth.     

Synthesis and herbicidal activity of optically active cinmethylin,its enantiomer,and C3-substituted cinmethylin analogs

We investigated the synthesis and herbicidal activity of optically active cinmethylin, its enantiomer, and C3-substituted cinmethylin analogs. Optically active cinmethylin could be obtained in seven steps with the Sharpless asymmetric dihydroxylation of α-terpinene. The synthesized cinmethylin and its enantiomer showed similar herbicidal activity, which was independent of the stereochemistry. Next, we synthesized cinmethylin analogs with various substituents at the C3 position. We found that analogs with methylene, oxime, ketone, or methyl groups at the C3 position show excellent herbicidal activity.     

Microscopic detection of reactive oxygen species generation in the compatible and incompatible interactions of Alternaria alternata Japanese pear pathotype and host plants

 Reactive oxygen species (ROS) generation was examined in the interaction of Alternaria alternata Japanese pear pathotype and host plants using three methods: nitro blue tetrazolium (NBT) method for microscopic detection of O₂ ⁻, diaminobenzidine (DAB) methods for microscopic detection of H₂O₂, and cerium chloride methods for ultrastructural detection of H₂O₂. ROS generation was detected by NBT and DAB methods at appressoria on leaves of susceptible cultivars and heat-shocked leaves of resistant cultivars but not in leaves of resistant cultivars. Ultrastructural detection by the cerium chloride method identified ROS generation at cell walls of appressoria and penetration pegs in susceptible, resistant leaves and heat-shocked leaves. These differences in the ultrastructural and microscopic data in resistant areas were due to the restriction of ROS generation in limited areas, the side facing the plant surface, of appressoria and penetration pegs. Therefore, ROS generation was apparently induced regardless of the resistance or susceptibility of the cultivar with the difference being in the volumes generated. After evaluating the pathological role of ROS generation in fungal structures, such generation was found to be associated with early penetration of cell walls in pear plants. Additionally, ROS generation in plants was also found in degrading pectin layers near infected hyphae and in plasma membrane modification sites in susceptible leaves but not in resistant leaves. ROS generation in susceptible leaves might be accompanied with plasma membrane damage, although the role of ROS generation in the pectin layers is not clear. ROS generation in both fungal and plant cells during their interaction was likely associated with the expression of susceptibility. Received: June 3, 2002 / Accepted: July 31, 2002     

Development of a Simple Method for Evaluation of Water Absorption Rate and Capacity of Rice Flour Samples

A simple method employing a commercially available canister was developed for the determination of mode of water absorption of rice flour samples. The samples prepared by four different grinding methods were used to analyze water absorption. The total amount of water in a flour sample was described by using an exponential model. Capacity and rate of water absorption of the samples were determined, and the relationship to baking quality of partially substituted rice bread was investigated. The water absorption was highly dependent on the method of grinding. Flours produced by wet jet‐milling of the grains, which absorbed a small amount of water at high speed, were most suitable for rice bread. The method was applicable to other food powders, provided that flour particles do not stick together or swell immediately upon contact with water.