Alteration of substrate specificity of cholesterol oxidase from Streptomyces sp. by site-directed mutagenesis

Despite the structural similarities between cholesterol oxidasefrom Streptomyces and that from Brevibacterium, both enzymesexhibit different characteristics, such as catalytic activity,optimum pH and temperature. In attempts to define the molecularbasis of differences in catalytic activity or stability, substitutionsat six amino acid residues were introduced into cholesteroloxidase using site-directed mutagenesis of its gene. The aminoacid substitutions chosen were based on structural comparisonsof cholesterol oxidases from Streptomyces and Brevibacterium.Seven mutant enzymes were constructed with the following aminoacid substitutions: L117P, L119A, L119F, V145Q, Q286R, P357Nand S379T. All the mutant enzymes exhibited activity with theexception of that with the L117P mutation. The resulting V145Qmutant enzyme has low activities for all substrates examinedand the S379T mutant enzyme showed markedly altered substratespecificity compared with the wild-type enzyme. To evaluatethe role of V145 and S379 residues in the reaction, mutantswith two additional substitutions in V145 and four in S379 wereconstructed. The mutant enzymes created by the replacement ofV145 by Asp and Glu had much lower catalytic efficiency forcholesterol and pregnenolone as substrates than the wild-typeenzyme. From previous studies and this study, the V145 residueseems to be important for the stability and substrate bindingof the cholesterol oxidase. In contrast, the catalytic efficiencies(k_cat/K_m) of the S379T mutant enzyme for cholesterol and pregnenolonewere 1.8- and 6.0-fold higher, respectively, than those of thewild-type enzyme. The enhanced catalytic efficiency of the S379Tmutant enzyme for pregnenolone was due to a slightly high k_catvalue and a low K_m value. These findings will provide severalideas for the design of more powerful enzymes that can be appliedto clinical determination of serum cholesterol levels and assterol probes.     

Biodegradation of poly(lactic acid)—cassava bagasse composites produced by injection molding

Neat poly (lactic acid) (PLA) and PLA/cassava bagasse (CB) composites were used to produce seedling tubes by extrusion and injection molding. The tubes were buried in simulated soil, and their biodegradation was investigated by weight loss, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR). After 180 days, the composites' biodegradation was higher than neat PLA material, and the higher the CB content, the higher the biodegradation, which caused fissures and voids in the material. The biodegradation of PLA/CB composites increased the phosphorus content in the soil after 180 days. Composites of PLA with CB, an abundant agro-industrial residue in Brazil, are promising because they can reduce the environmental impact due to CB's proper destination, and the composites' costs and biodegradation are faster than pure PLA material. Both the faster biodegradation of the tube and the higher P content are advantageous for seedling tubes.     

Internal oxidation of dilute silver alloys

Internal oxidation of dilute silver alloys containing Al, Mg, Zn, Cu, and Sn was studied in air at temperatures between 573 K and 1173 K. Electrical resistivity, gravimetric, and gas-extraction measurements were made. The general trend of the resistivity is that it increases upon oxidation at lower temperatures and the resistivity decreases at higher temperatures in all of these alloys except Ag-Mg, in which it increases even at 1173 K. The increase in resistivity is considered to be related to the formation of clusters having excess oxygen. A detailed investigation was performed on Ag-Al alloys. The O/Al ratio in the clusters in Ag-2.2 at.% Al is much higher on oxidation at 773 K than for stoichiometric Al₂O₃ at 1173 K. The clusters release the excess oxygen on subsequent annealing at high temperatures, and decompose to stable Al₂O₃ at 1173 K.     

Kinetic roughening transition and missing regime transition of melt crystallized polybutene-1 tetragonal phase: growth kinetics analysis

The morphology and lateral growth rate of isotactic polybutene-1 (it-PB1) have been investigated for crystallization from the melt over a wide range of crystallization temperatures from 50 to 110°C. The morphology of it-PB1 crystals is a rounded shape at crystallization temperatures lower than 85°C, while lamellar single crystals possess faceted morphology at higher crystallization temperatures. The kinetic roughening transition occurs around 85°C. The nucleation and growth mechanism for crystallization does not work below 85°C, since the growth face is rough. However, the growth rate shows the supercooling dependence derived from the nucleation and growth mechanism. The nucleation theory seems still to work even for rough surface growth. Possible mechanisms for the crystal growth of this polymer are discussed.     

Thermal behaviors and flame‐retardancy of styrene–ethylene–butadiene–styrene–block copolymer containing various additives

The thermal behaviors and the flame‐retardancy of styrene–ethylene–butadiene–styrene–block copolymer containing various additives were studied. The combustion was measured by the Underwriter laboratory (UL) test and cone calorimeter test and thermogravimetric analysis and program‐mass spectroscopy were applied to analyze the thermal behaviors. The blend with halogen additives showed the best result in the UL test. However, the blend with red‐phosphorous was the best in the cone calorimeter test. As the styrene sequence in the copolymer tended to degradate at a lower temperature, the major scission products spouted out from the polymer surface originated from polystyrene. The shorter the ignition times of the blends with red‐phosphorous were, the lower the peak heat release rates were. It was an interesting phenomenon because it suggested that the chemical structure of the residue changed to more stable polymers. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 104: 156–161, 2007     

A novel spherical carbon

We developed a novel spherical carbon material. The spherical carbon is composed of a high density of carbon nanotubes or nanofilaments, and includes an oxidized diamond particle as a core. Syntheses of this carbon in high volume with high selectivity may be possible. It is expected that this carbon will be useful as a catalyst material for fuel cells, electric double-layer capacitors, etc.     

Comparison of CTOD standards: BS 7448-Part 1 and revised ASTM E1290

Crack tip opening displacement (CTOD) has been calculated using the plastic hinge model with an assumed rotational center since the British Standards Institution (BS) standardized BS5762 in 1979. The American Society for Testing and Materials (ASTM) accepted the plastic hinge model and standardized E1290 in 1989. However, ASTM revised E1290 in 2002, and has proposed a conversion from J to CTOD. CTOD-based fracture toughness evaluation has been widely used for the defect assessment of many welded structural components, and two different CTOD calculations could lead to confusion for Fitness-for-Service. In this study, the effects of CTOD testing methodologies on CTOD values were investigated according to round robin tests conducted by the Japan Welding Engineering Society (WES), and the concept of CTOD as a fracture parameter is discussed.     

Layered mixed-anion compounds: Epitaxial growth,active function exploration,and device application

Optoelectronic properties and device applications of layered mixed-anion compounds such as oxychalcogenide LaCuOCh (Ch = chalcogen) and oxypnictide LaT_MOPn (T_M = 3d transition metal, Pn = pnicogen) are reviewed. Several distinctive functions have been found in these materials based on our original material exploration concept. Fabrication of high-quality epitaxial films of LaCuOCh leads to clarifying the excellent electrical and optical properties such as high hole mobility of 8 cm²/(V s) and heavy hole doping at >10²¹ cm^?3 in LaCuOSe, and sharp and tunable-wavelength photoluminescence in the solid–solution systems in LaCuOCh. In addition, a room temperature operation of a light-emitting diode is demonstrated using LaCuOSe as a light-emitting layer. These results suggest that the layered oxychalcogenides have potential for light-emitting layers as well as transparent hole-injection layers in organic/inorganic light-emitting diodes. Furthermore, by extending the material system from the copper-based oxychalcogenides to isostructural compounds, transition metal-based oxypnictides LaT_MOP (T_M = Fe, Ni), we have found novel superconductors, LaFeOP and LaNiOP.     

Fundamental Behavior of Direct-Coupled Submicrosecond Pulse Energization in Electrostatic Precipitators

The fundamental characteristics of the direct-coupled pulse energization in electrostatic precipitators (EP's) are investigated in a test rig comprised of a coal pulverizing facility, a furnace, a gas cooling system, and an EP. The EP consisted of a corona electrode with barbed wires and a collection electrode with 300-mm duct spacing. This simulates the type of EP normally used in a coal-fired power station. The pulse-energized collection field shows a very high collection performance of more than 99 percent collection efficiency and the performance enhancement compared to the conventional dc energization, causing very severe back corona, becomes as large as H = 2.50.H is the enhancement factor in terms of the Deutsch migration velocity in the case of the very high resistivity dust of rd = 2 x 1013 ?cm at the gas temperature of Tg = 150°C. Even at the lower level of the dust resistivity at Tg = 110°C, where no back corona occurs in the dc-energized collection field, the performance enhancement is as large as H = 1.21. The mechanism of such great performance enhancement is likely to be the result of the very uniform and very intense formation of corona plasmas on the wires as a result of the very fast rise in the pulse voltage applied and a special sawtooth waveform of the operating voltage between the corona and collection electrodes.