Hydrolysis of beta-glucosyl ester linkage of p-hydroxybenzoyl beta-D-glucose, a chemically synthesized glucoside, by beta-glucosidases

To investigate the hydrolysis of glucosyl esters by beta-glucosidase, p-hydroxybenzoyl beta-D-glucose (pHBG) was chemically synthesized. The hydrolytic activity of some beta-glucosidases for pHBG was compared to that for p-nitrophenyl beta-glucoside (pNPG). The Clavibacter michiganense and Flavobacterium johnsonae enzymes could hydrolyze pHBG and steviol glycosides which are natural glucosyl esters. The commercial beta-glucosidase originating from Caldocellum saccharolyticum also hydrolyzes pHBG despite having no activity for steviol glycosides. The beta-glucosidase from Aspergillus niger cleaved the glucosyl ester linkage much more weakly than the glucosidic linkage. The pH-activity profile for the hydrolysis of pHBG was similar to that of pNPG by the C. saccharolyticum beta-glucosidase. The similar profiles for these substrates suggested that the active site for the glucosyl ester chemically resembles that for glucoside with respect to catalysis. Kinetic analysis of the C. saccharolyticum beta-glucosidase for mixed substrates of pHBG and pNPG showed that the hydrolysis of pHBG competed with that of pNPG. This result indicated that there is only one active site for both the glucosyl ester and glucoside. Mass spectroscopic analysis of the hydrolysates of pHBG in H218O suggested that beta-glucosidase hydrolyzes glucosyl esters between the anomeric carbon of glucose and the carbonyl oxygen, not between the carbonyl carbon and the carbonyl oxygen.     

On-line derivative estimation for the multiclass single-serverpriority queue using perturbation analysis

Gradient estimation via smoothed perturbation analysis (SPA) is considered for the multiclass single-server priority queue. Although the problem like this requires additional subpaths, generally it is shown that the estimators can be derived without any additional sample paths     

Thin oxide films of silicon by high pressure oxidation

High pressure oxidation of silicon was performed at an oxidation temperature range of 650 to 800°C for thin oxide films with about 300 å thickness. The index of refraction was dependent on an oxidation temperature but independent of the oxidation pressure as 1.475 at 750°C. The dielectric breakdown strength of the oxide film was measured by the voltage ramping,method. The fixed oxide charge about 1.0 × 10¹¹ cm^?2 was also measured from the high frequency C-V measurement. The pulse scanning C-V measurement technique was used to measure the minority carrier generation rate in the depleted surface. The surface generation velocity was slightly dependent on the oxidation temperature and indicated that the fast surface states increased with decreasing oxidation temperature.     

Improvement of bitumen-waste product in leachability—Leachability of bitumen product containing BWR's evaporator concentrate

The method has been developed to improve the bitumen product which incorporates an evaporator concentrate from a BWR, with respect to the swelling and leaching. The leachability of the product has been measured by the method recommended by the IAEA. The swelling of the product is successfully prevented by the addition of calcium chloride. The specimen containing the waste up to Wa/B (Weight ratio of Na₂SO₄ + CaCl₂ and bitumen) = 60:40 shows no pronounced swelling, when it is immersed in water. The cumulative fractions of ¹³⁷Cs and ⁶⁰Co leached from a specimen which does not contain CaCl₂ are 0.65 and 0.2 at the leaching time of 30 days. On the hand, the corresponding value at 100 days for the specimen with calcium chloride addition is 5 × 10^?4 for ¹³⁷Cs and 1 × 10^?4 for ⁶⁰Co. The coating of the specimen surface with a fresh bitumen (5 mm thickness) reduces the leachability further. These results indicate that this method is effective to improve the bitumen product incorporating BWR's evaporator concentrate.     

Composite of Au nanoparticles and molecularly imprinted polymer as a sensing material

A molecularly imprinted polymer with immobilized Au nanoparticles (Au-MIP) is reported as a novel type of sensing material. The sensing mechanism is based upon the variable proximity of the Au nanoparticles immobilized in the imprinted polymer, which exhibits selective binding of a given analyte accompanied by swelling that causes a blue-shift in the plasmon absorption band of the immobilized Au nanoparticles. Using adrenaline as the model analyte, it was shown that molecular imprinting effectively enhanced the sensitivity and selectivity, and accordingly, Au-MIP selectively detects the analyte at 5 microM. The combination of molecular imprinting and the Au nanoparticle-based sensing system was shown to be a general strategy for constructing sensing materials in a tailor-made fashion due to wide applicability of the imprinting technique and the independence of the sensing mechanism from the analyte recognition system.     

Dose evaluation in criticality accident conditions using transient critical facilities fueled with a fissile solution

Neutron dose measurement and evaluation techniques in criticality accident conditions using a thermo luminescence dosemeter (TLD) was studied at the Transient Experiment Critical Facility (TRACY) of Japan Atomic Energy Research Institute (JAERI). In the present approach, the absorbed dose is derived from the ambient dose equivalent measured with a TLD, using the appropriate conversion factor given by computation. Using this technique, the neutron dose around the SILENE reactor of the Institute for Radioprotection and Nuclear Safety (IRSN) of France was measured in the Accident Dosimetry Intercomparison Exercise (June 10-21, 2002) organized by OECD/NEA and IRSN. In this exercise, the gamma dose was also measured with a TLD. In this report, measurements and evaluation results at TRACY and SILENE are presented.     

Purification, bacteriolytic activity, and specificity of beta-lytic protease from Lysobacter sp. IB-9374

Lysobacter sp. IB-9374, which was isolated from soil as a high lysyl endopeptidase-producing strain (Chohnanet al., FEMS Microbiol. Lett., 213, 13-20, 2002), was found to produce a beta-lytic protease capable of lysing gram-positive bacteria such as Staphylococcus aureus, Microccocuseus, and Bacillus subtilis. The Lysobacter strain secreted the beta-lytic protease into the culture medium at a 2.4-fold higher level than Achromobacter lyticus. The enzyme was highly purified through a series of six steps with a high yield. The enzyme was strongly inhibited by tetraethylene-pentamine and 1,10-phenanthroline. The purified enzyme lysed more efficiently almost all the gram-positive bacteria tested than lysozyme, lysostaphin, and mutanolysin. The enzyme was very similar to Achromobacter beta-lytic protease containing one zinc atom in terms of amino acid composition and N-terminal sequence. The nucleotide sequence revealed that the mature enzyme was composed of 179 amino acid residues with additional 198 amino acids at the amino-terminal end of the enzyme. The deduced amino acid sequence of the mature enzyme coincided with that of the Achromobacter enzyme, although the prepro-region showed a 41% sequence identity with the counterpart. These results indicate that Lysobacter sp. is a useful strain for an efficient large-scale preparation of beta-lytic protease capable of lysing bacteria.     

Chemiluminescence detection for a microchip capillary electrophoresis system fabricated in poly(dimethylsiloxane)

Chemiluminescence (CL) detection integrated with a microchip capillary electrophoresis (MCE) system that was fabricated in poly(dimethylsiloxane) was demonstrated for chemical and biochemical analyses. Two model CL systems were involved here: metal ion-catalyzed luminol-peroxide reaction and dansyl species conjugated peroxalate-peroxide reaction. Different strategies based on three chip patterns (cross, cross combining with Y, and cross combining with V) to perform on-line CL detection for MCE were evaluated and compared in terms of sensitivity, reproducibility, and peak symmetry. The chip pattern of cross combining with Y proved to be promising for the luminol-peroxide CL system, while the chip pattern of cross combining with V was preferred for the peroxalate-peroxide system where CL reagent could not be effectively transported by electroosmotic flow. A detection limit down to submicromolar concentrations (midattomole) was achieved with good reproducibility and symmetric peak shape. Successful separation of three metal cations such as Cr(III), Co(II), and Cu(II) and chiral recognition of dansyl phenylalanine enantiomers within 1 min revealed distinct advantages of combining MCE with CL detection for rapid and sensitive analyses.     

Mechanistic study on analyte focusing by dynamic pH junction in capillary electrophoresis using computer simulation

Dynamic pH junction is an on-line preconcentration method in capillary electrophoresis (CE) based on electrokinetic focusing of weakly ionic analytes with in large sample volumes in a multisection electrolyte system. In this report, experiments and computer simulations were performed to gain a better insight of the analyte focusing mechanism when a dynamic pH junction was used. A computer program, SIMUL, was used to simulate the band-narrowing process of a group for phenol derivatives under optimized buffer conditions, which were compared with experimental results. Computer simulations revealed the formation of a sharp moving pH boundary within the sample zone causing efficient focusing of long plugs of weakly acidic analytes based on their pKa. These studies offered useful information for understanding the band-narrowing process by control of the depth and lifetime of the moving pH boundary as a function of analyte pKa, sample pH, and injection length. The change in pH of the sample within the capillary was also estimated by measuring the absorbances of an analyte at two different wave-lengths. Optimization of analyte focusing resulted in enhanced detection responses of about 60-450-fold in terms of peak heights for some phenol derivatives' relation to conventional injections. Dynamic pH junction represents a novel approach to control band dispersion (peak width) and selectivity (mobility) of specific analytes for high-resolution CE separations.