A lysine to arginine substitution at position 146 of rabbit aldolase A changes the rate-determining step to Schiff base formation

Lys146 of rabbit aldolase A [D-fructose-1,6-bis(phosphate):D-glyceraldehyde-3-phosphate lyase, EC 4.1.2.13 [EC] ] was changedto arginine by site-directed mutagenesis. The k_cat of the resultingmutant protein, K146R, was 500 times slower than wild-type insteady-state kinetic assays for both cleavage and condensationof fructose-1,6-bis(phosphate), while the Km for this substratewas unchanged. Analysis of the rate of formation of catalyticintermediates showed K146R was significantly different fromthe wild-type enzyme and other enzymes mutated at this site.Single-turnover experiments using acid precipitation to trapthe Schiff base intermediate on the wild-type enzyme failedto show a build-up of this intermediate on K146R. However, K146Rretained the ability to form the Schiff base intermediate asshown by the significant amounts of Schiff base intermediatetrapped with NaBH₄. In the single-turnover experiments it appearedthat the Schiff base intermediate was converted to productsmore rapidly than it was produced. This suggested a maximalrate of Schiff base formation of 0.022 s^–1, which wasclose to the value of k_cat for this enzyme. This observationis strikingly different from the wild-type enzyme in which Schiffbase formation is >100 times faster than k_cat. For K146Rit appears that steps up to and including Schiff base formationare rate limiting for the catalytic reaction. The carbanionintermediate derived from either substrate or product, and theequilibrium concentrations of covalent enzyme-substrate intermediates,were much lower on K146R than on the wild-type enzyme. The greaterbulk of the guanidino moiety may destabilize the covalent enzyme-substrateintermediates, thereby slowing the rate of Schiff base formationsuch that it becomes rate limiting. The K146R mutant enzymeis significantly more active than other enzymes mutated at thissite, perhaps because it maintains a positively charged groupat an essential position in the active site or perhaps the Argfunctionally substitutes as a general acid/base catalyst inboth Schiff base formation and in subsequent abstraction ofthe C4-hydroxyl proton.     

Solid extraction of caffeine and theophylline from green tea by molecular imprinted polymers

This paper involves a feasibility study on using molecular imprinted polymers as the sorbent materials in solid phase extraction for caffeine and theophylline from green tea. Two kinds of MIPs, with caffeine-theophylline mixture and pentoxifylline-theophylline mixture as the templates respectively, MAA as the monomer, EDMA as the crosslinker and ATBN as the initiator, were applied to this purpose. Mixture solution of caffeine and theophylline (1 Μg/ ml in acetonitrile) was applied to the solid extraction cartridges following a load, wash and elute procedure with acetonitrile, methanol, methanol-acetic acid (90/10, v/v) as the solvents, respectively. This solid phase extraction protocol was applied for extraction of caffeine and theophylline from green tea. Comparison between the results obtained with the MIPs cartridges and a traditional C₁₈ reversed-phase cartridge was made. It showed that the MIP-based sorbent on the solid phase extraction was comparable with that of C₁₈ material. HPLC analysis using a C₁₈ column (5 Μm, 250× 4.6 mm from Rstech corporation), methanol: water (60 :40, v/v) as the mobile phase at a flow rate of 0.6 ml/min was applied for the quantitative determination.     

Numerical study of coal particle cluster combustion under quiescent conditions

Behavior of ignition and combustion of coal particle cluster under a quiescent condition was numerically simulated by solving balance equations of mass and enthalpy with combustion kinetic models of volatiles and char. Two-flame structure, one flame penetrating into the cluster and the other moving out of the cluster, was predicted during the combustion of coal particle cluster. Effects of radiative heat transfer, group number, ambient temperature, coal particle size, and oxygen concentration on ignition and combustion of coal particle clusters were also analyzed. Simulations indicated that the gas volume fraction of coal particle cluster increases with time after devolatilization. Gas velocity passing through the cluster surface varied significantly at volatile liberation. The ignition time delay was reduced with the increase of ambient temperature. The cluster devolatilization rate and char burning rate increased while the ignition time delay decreased with the increase of ambient oxygen concentration.     

Mobile phase composition for resolving whey proteins in reversed-phase high performance liquid chromatography

Reversed-phase high-performance liquid chromatography was successfully developed for the simultaneous and rapid separation for the main whey proteins, α-Lactalbumin and β-Lactoglobulin. This method consisted of a linear gradient of the two mobile phases of 0.1% trifluoroacetic acid in water and 0.1% trifluoroacetic acid in acetonitrile. The total run time for this separation was approximately 30 min, and α-Lactalbumin was eluted followed byβ- Lactoglobulin. The injection volume was fixed at 20 μl and the flow rate was 1 ml 1/min. The optimum mobile phase composition and gradient conditions to separate α-Lactalbumin and β-Lactoglobulin (A+B) were experimentally obtained at the 15 μm particle with a pore size of 300 Å on the linear-gradient mode.     

Effect of Antimony Addition on Electrical and Optical Properties of Tin Oxide Film

Antimony-doped tin oxide films were prepared on Corning glass substrate by chemical vapor deposition from a gas mixture of SnCl₄–SbCl₅–H₂O. The electrical conductivity and optical transmission of tin oxide films were studied with antimony doping. The film conductivity increased largely without losing optical transparency with a small addition of antimony. The increase of the conductivity was attributed mainly to more electrons donated by pentavalent Sb ions in the SnO₂ lattice. Large additions of antimony, however, diminished the conductivity, optical transparency, and crystallinity of the film. The diminution was found to be caused by fine Sb₂O₅ phases codeposited with SnO₂.     

Desulfurization in a plate-type gas-lift photobioreactor using light emitting diodes

Hydrogen sulfide gas was removed in a 2-dimensional gas-lift reactor by the photosynthetic microorganismChlorobium thiosulfatophilum using light emitting diodes (LEDs) as a light source. LEDs saved light energy by 99% compared with the incandescent light source. The plate-type gas-lift reactor removed hydrogen sulfide five times better per unit mg of protein, and performed two times better in the maximum performance per unit luminous flux, compared with cylindrical fermentors.     

Influence of Potassium on Preparation and Performance of PTC Resistors

Different amounts of K₂CO₃ were added to (Ba,Sr)TiO₃-based PTCR (positive temperature coefficient of resistance) ceramics to investigate their influence on the microstructural and electrical properties. Experimental results showed that the incorporation of K acted as an A-site acceptor-type dopant. In addition to enhancing discontinuous grain growth, the increase of K₂CO₃ was found to raise the room-temperature resistivity which was dominated by grain-boundary resistance rather than grain resistance. By adjusting to a suitable amount of donor dopant, the inherent contamination of K in raw material can be compensated to achieve a high-quality PTC resistor.     

An order-picking operations system for managing the batching activities in a warehouse

Nowadays, customer orders with high product variety in small quantities are often received and requested for timely delivery. However, the order-picking process is a labour-intensive and costly activity to handle those small orders separately. In such cases, small orders are often grouped into batches so that two or more orders can be served at once to increase the picking efficiency and thus reduce the travel distance. In this paper, an order-picking operations system (OPOS) is proposed to assist the formulation of an order-picking plan and batch-handling sequence. The study integrates a mathematical model and fuzzy logic technique to divide the receiving orders into batches and prioritise the batch-handling sequence for picking, respectively. Through the proposed system, the order-picking process can be managed as batches with common picking locations to minimise the travel distance, and the batch-picking sequence can be determined as well. To demonstrate the use of the system, a case study in a third-party logistics warehouse is presented, and the result shows that both the order-picking activity and labour utilisation can be better organised.     

Effect of reactive diluents on properties of unsaturated polyester/montmorillonite nanocomposites

Unsaturated polyester (UP)/montmorillonite (MMT) nanocomposite was prepared by using hydroxypropylacrylate (HPA) as a reactive diluent instead of conventional styrene monomer and the effect of polarity of reactive diluent on properties of nanocomposite was investigated. X‐ray and mechanical test data indicated that mixing for an extended period of time is essential to enhance the physical properties of nanocomposites in the UP/Cloisite 6A system. This was attributed to the high polarity of HPA that may disturb the preintercalation of UP resin into the galleries of MMT. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 238–242, 2004     

Removal of formaldehyde at low concentration using various activated carbon fibers

Adsorption of low concentration formaldehyde on pitch‐based, rayon‐based, and PAN‐based activated carbon fibers (ACFs) and an unactivated PAN‐based carbon fiber (PAN‐CF) was investigated by a dynamic method. The pore structure and surface chemistry of these samples were characterized by liquid nitrogen adsorption, elemental analysis, and X‐ray photoelectron spectroscopy. Results revealed that the pore structure, especially surface chemical composition, greatly influence the formaldehyde adsorption. PAN‐based ACFs showed the highest formaldehyde adsorption capacity because there are more abundant nitrogen‐containing groups, especially pyrrolic, pyridonic, pyridinic, and quaternary on the surface. The breakthrough time and formaldehyde adsorption capacity of one kind of PAN‐ACF were 361 min and 0.478 mmol/g, respectively. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2007