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Imoto Taiji; Ueda Tadashi; Tamura Tomohiro; Isakari Yoshimasa; Abe Yoshito; Inoue Makoto; Miki Takeyoshi; Kawano Keiichi; Yamada Hidenori 《Protein engineering, design & selection : PEDS》1994,7(6):743-748
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 kcat 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. 相似文献
23.
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. 相似文献
24.
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. 相似文献
25.
Yujiro Watanabe Yusuke Moriyoshi Yasushi Suetsugu Toshiyuki Ikoma Takeshi Kasama Tadashi Hashimoto Hirohisa Yamada Junzo Tanaka 《Journal of the American Ceramic Society》2004,87(7):1395-1397
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 PO4 3− ions in (NH4 )3 PO4 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. 相似文献
26.
Maeda Yoshitake; Koga Hiroshi; Yamada Hidenori; Ueda Tadashi; Imoto Taiji 《Protein engineering, design & selection : PEDS》1995,8(2):201-205
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, 27902797].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. 相似文献
27.
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. 相似文献
28.
29.
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. 相似文献
30.
Yoshifumi Matsumoto Masaru Setomoto Daiji Noda Tadashi Hattori 《Microsystem Technologies》2008,14(9-11):1373-1379
The demand for microactuators is increasing recently. The key technology to realizing practical microactuators is microfabrication process. In the production of microminiature components, the technologies for processing high-aspect-ratio structures are essential. As one of these technologies, the LIGA process is widely known. Our laboratory researches the LIGA process to three-dimensional microfabrication and established the cylindrical-microcoil production process. In this paper, we have fabricated the cylindrical-microcoil for the solenoidal electromagnetic type microactuator. We designed and analyzed microactuators, and fabricated and evaluated microactuator coils produced by the combination of three-dimensional X-ray lithography and level copper plating. We succeeded in creating threaded groove-shaped structures with 10 μm line width, 20 μm pitch, and aspect ratio of 5 on the surface of an acrylic pipe by means of three-dimensional X-ray lithography. As a measure to suppress void generation, which is one of the shortcomings of electrolytic plating processes, the sputtering apparatus and plating equipment were improved, a pretreatment process was additionally provided, and the actual electrolytic plating method was improved. As a result, a void-free metallic deposit could be formed on a thin coil line. The processing technology enables the formation of thin-wire coil lines whose current paths feature a large allowable current-carrying capacity, enabling the production of miniature, high-output microactuators. 相似文献