Lysozyme requires fluctuation of the active site for the manifestation of activity

Mutations around His15 which lie far away from the active site,stimulated glycol chitin activity of lysozyme at physiologicaltemperature. Del-Argl4Hisl5 lysozyme, a mutant lysozyme whoseArgl4 and Hisl5 were deleted together, and has the highest activityamong these mutant lysozymes, had a similar binding abilityto a trimer of N-acetyl-glucosamine, a substrate analogue, relativeto native lysozyme. This suggests that the increased activitywas due to an increased k_cat in the catalysis reaction. TheH-D exchange rate of the N-1 proton in the Trp63 which is locatedin the active site cleft, was enhanced in the Del-Argl4Hisl5lysozyme, while 2-D proton NMR analysis revealed no conformationalchange around Trp63. We conclude that some sort of fluctuationat the active site might be required for the manifestation ofactivity. This theory is supported by the finding that the Del-Argl4Hisl5lysozyme showed a shift in temperature dependency of activityto lower temperatures compared with that of native lysozyme.     

pH-controlled uphill transport of ammonium ions through polymer membranes with sulfonic acid groups

Uphill transport of ammonium ions through a membrane with sulfonic acid groups were investigated with pH-controllers which keep the solution at a constant pH. A membrane with sulfonic acid groups was prepared by casting an aqueous solution containing poly(styrenesulfonic acid) and poly(vinyl alcohol) on a glass plate. When a membrane with one side alkaline and the other acidic was fixed as a diaphragm in a cell, ammonium ions were transported from the alkaline side to the acidic side through the membrane against the concentration gradient of the adjacent solutions. Uphill transport of ammonium ions with pH-controllers was more efficient than without, thus keeping the pH difference between both sides of the membrane constant, which is a driving force for the uphill transport. Furthermore, the effect of pH of the acidic side on the uphill transport was investigated and the mechanism of the pH-controlled uphill transport is discussed.     

Studies on syntheses and permeabilities of special polymer membranes

Summary The permeabilities of various cellulose ester membranes and cellulose membranes were investigated using pure water and aqueous solution of glycerol as feed. In all membranes, the permeation rate for aqueous solution of glycerol was greater than for pure water. The permeation rates of cellulose ester membranes increased with an increase in number of carbon atoms of the substituent group in cellulose ester.     

Hydrothermal Formation of Hydroxyapatite Layers on the Surface of Type-A Zeolite

Type-A zeolite evenly covered with hydroxyapatite thin layers was prepared using hydrothermal treatment at 120°C for 8 h under autogenous pressure. The hydroxyapatite needlelike nanocrystals, 100–200 nm in diameter and 30 nm in thickness, were grown under the reaction between discharged Ca²⁺ ions from type-A zeolite and PO₄³⁻ ions in (NH₄)₃PO₄ solution. The preferential orientations of the c -axis of hydroxyapatite crystals perpendicular to a zeolite surface were observed using transmission electron microscopy. The crystal structure of type-A zeolite was not destroyed under the reaction, but the surface morphology was changed only with complete covering of scaly hydroxyapatite particles.     

Transient analysis of the release and reduction of NOx using a Pt/Ba/Al2O3 catalyst

The release and reduction of NO_x in a NO_x storage-reduction (NSR) catalyst were studied with a transient reaction analysis in the millisecond range, which was made possible by the combination of pulsed injection of gases and time resolved time-of-flight mass spectrometry. After an O₂ pulse and a subsequent NO pulse were injected into a pellet of the Pt/Ba/Al₂O₃ catalyst, the time profiles of several gas products, NO, N₂, NH₃ and H₂O, were obtained as a result of the release and reduction of NO_x caused by H₂ injection. Comparing the time profiles in another analysis, which were obtained using a model catalyst consisting of a flat 5 nmPt/Ba(NO₃)₂/cordierite plate, the release and reduction of NO_x on Pt/Ba/Al₂O₃ catalyst that stored NO_x took the following two steps; in the first step NO molecules were released from Ba and in the second step the released NO was reduced into N₂ by H₂ pulse injection. When this H₂ pulse was injected in a large amount, NO was reduced to NH₃ instead of N₂.

A only small amount of H₂O was detected because of the strong affinity for alumina support. We can analyze the NO_x regeneration process to separate two steps of the NO_x release and reduction by a detailed analysis of the time profiles using a two-step reaction model. From the result of the analysis, it is found that the rate constant for NO_x release increased as temperature increase.     

Effective renaturation of reduced lysozyme by gentle removal of urea

To increase the folding yield of concentrated reduced lysozyme,we developed a renaturation method by means of dialysis fromconcentrated urea with redox agents. After lysozyme was incubatedin the reducing buffer (8 M urea solution) with oxidized glutathione,renaturation of reduced lysozyme was started by dialysis againstthe dialyzing buffer containing 8 M urea with redox agents.The urea concentration of the dialyzing bottle was graduallydiluted with dialyzing buffer without urea at a flow rate of0.1 ml/min by high pressure pump. Using this systematic dialysis,a concentration as high as 5 mg/ml of reduced lysozyme couldbe renaturated in 80% yield, while the folding yield was <5%even at a concentration of 1 mg/ml using a conventional rapiddilution method [Goldberg et al. (1991) Biochemistry, 30, 2790–2797].Therefore, it was concluded that gentle removal of urea fromdenatured proteins, dissolved in concentrated urea solution,by means of dialysis should be useful to renature denaturedproteins effectively.     

Self-assembled monolayers of p-toluene and p-hydroxymethylbenzene moieties adsorbed on iron by the formation of covalent bonds between carbon and iron atoms for protection of iron from corrosion

An iron electrode was modified by electrolytic reduction in deaerated acetonitrile solution of p-toluenediazonium tetrafluoroborate CH₃C₆H₄N₂BF₄ (TDFB) or p-hydroxymethylbenzenediazonium tetrafluoroborate HOCH₂C₆H₄N₂BF₄ (HOTDFB) below 10 °C to form a self-assembled monolayer (SAM) of toluene CH₃C₆H₄- or hydroxymethylbenzene HOCH₂C₆H₄- (HOMB) moiety, probably adsorbed on the electrode by the formation of a covalent bond between carbon and iron atoms, as shown in references. The protective ability of the layer was examined by polarization measurement of the electrode in an aerated 0.5 M NaCl solution. The protective efficiencies of these two SAMs were not high, around 30%, a little higher than that of the toluenethiol CH₃C₆H₄SH SAM which was anchored on iron via a coordinate bond between sulfur and iron atoms. The iron surfaces modified with TDFB and HOTDFB were characterized by contact angle measurement, FTIR reflection spectroscopy and X-ray photoelectron spectroscopy. The persistence in protection of iron against corrosion by coverage with the HOMB SAM was confirmed by polarization measurements after immersion in 0.5 M NaCl for a long period of the time.     

超声波功率对铸锭内的气孔及组织细化的影响

研究了超声波功率对铝合金铸锭内气孔的影响,同时也考察了超声波功率对铝合金铸锭组织细化的影响,分析了超声波对铝合金铸锭内气孔影响的原因。研究结果表明,超声波在金属熔体内产生的空化效应可分为亚空化效应阶段和发展-发达阶段,只有在发展-发达阶段才具有良好的除气效果。     

Activity measurement of gallium in the B2 phase of the CoGa–Sb system by the EMF method with zirconia-based solid electrolyte

Gallium activity in the B2 phase regions of both binary Co–Ga and ternary Co–Ga–Sb systems was measured by EMF method with stabilized zirconia solid electrolyte The temperature range was 1073–1273 K and Sb concentrations were 1, 2 and 3 mol fractions. Ga activity at 1173 and 1273 K increases sharply in Ga rich region and the addition of Sb to the CoGa phase increases Ga activity. Activity change corresponds to the lattice parameter change with Sb addition to the CoGa phase.