Single amino acid substitutions can further increase the stability of a thermophilic L-lactate dehydrogenase

Lactate dehydrogenases are of considerable interest as stereospecificcatalysts in the chemical preparation of enantiomerically pure-hydroxyacid synthons. For such applications in synthetic organicchemistry it would be desirable to have enzymes which tolerateelevated temperatures for prolonged reaction times, to increaseproductivity and to extend then applicability to poor substrates.Here, two examples are reported of significant thermostabilizations,induced by sitedirected mutagenesis, of an already thermostableprotein, the L-lactate dehydrogenase (EC 1.1.1.27 [EC] , 35 kDa permonomer subunit) from Bacillus stearothermophilus. Thermal inactivationof this enzyme is accompanied by irreversible unfolding of thenative protein structure. The replacement of Argl71 by Tyr stabilizesthe enzyme against thermal inactivation and unfolding. Thisstabilizing effect appears to be based on improved interactionsbetween the subunits in the core of the active dimeric or tetramericforms of the enzyme. The thermal stability of L-lactate dehydrogenasevariants with an active site Arg residue, either in the 171(wild-type) or in the 102 position, is further increased bysulfate ions. The two stabilizing effects are additive, as foundfor the Argl71Tyr/ Gln1O2Arg double mutant, for which the stabilityof the protein in 100 mM sulfate solution reaches that of L-lactatedehydrogenases from extreme thermophiles. All mutant proteinsretain significant catalytic activity, both in the presenceand absence of stnhilfoing salts, and are viable catalysts inpreparative scale reactions.     

Membranen in der Verfahrenstechnik

Membranes in chemical engineering . In the last 10 years membranes and membrane processes have evolved from a useful laboratory tool to an industrial product of significant technical and commercial impact. In some applications membrane processes have today not only replaced some conventional separation procedures in the chemical process industry, because they are often more economical and yield better quality products, but they have also successfully been utilized to solve mass separation problems where conventional procedures failed or are too expensive. This contribution discusses the state of the art of membranes and membrane processes as well as their major applications, with the main emphasis being placed on more recent developments.     

Clustering semantically heterogeneous distributed aggregate databases

Databases developed independently in a common open distributed environment may be heterogeneous with respect to both data schema and the embedded semantics. Managing schema and semantic heterogeneities brings considerable challenges to learning from distributed data and to support applications involving cooperation between different organisations. In this paper, we are concerned mainly with heterogeneous databases that hold aggregates on a set of attributes, which are often the result of materialised views of native large-scale distributed databases. A model-based clustering algorithm is proposed to construct a mixture model where each component corresponds to a cluster which is used to capture the contextual heterogeneity among databases from different populations. Schema heterogeneity, which can be recast as incomplete information, is handled within the clustering process using Expectation-Maximisation estimation and integration is carried out within a clustering iteration. Our proposed algorithm resolves the schema heterogeneity as part of the clustering process, thus avoiding transformation of the data into a unified schema. Results of algorithm evaluation on classification, scalability and reliability, using both real and synthetic data, demonstrate that our algorithm can achieve good performance by incorporating all of the information from available heterogeneous data. Our clustering approach has great potential for scalable knowledge discovery from semantically heterogeneous databases and for applications in an open distributed environment, such as the Semantic Web.     

A New Approach to Two-View Motion Segmentation Using Global Dimension Minimization

We present a new approach to rigid-body motion segmentation from two views. We use a previously developed nonlinear embedding of two-view point correspondences into a 9-dimensional space and identify the different motions by segmenting lower-dimensional subspaces. In order to overcome nonuniform distributions along the subspaces, whose dimensions are unknown, we suggest the novel concept of global dimension and its minimization for clustering subspaces with some theoretical motivation. We propose a fast projected gradient algorithm for minimizing global dimension and thus segmenting motions from 2-views. We develop an outlier detection framework around the proposed method, and we present state-of-the-art results on outlier-free and outlier-corrupted two-view data for segmenting motion.     

Moisture absorption into ultrathin hydrophilic polymer films on different substrate surfaces

Moisture absorption into ultrathin poly(vinyl pyrrolidone) (PVP) films with varying thickness was examined using X-ray reflectivity (XR) and quartz crystal microbalance (QCM) measurements. Two different surfaces were used for the substrate: a hydrophilic silicon oxide (SiO_x) and a hydrophobic hexamethyldisilazane (HMDS) treated silicon oxide surface. The total equilibrium moisture absorption (solubility) was insensitive to the surface treatment in the thickest films (≈150 nm). However, strong reductions in the equilibrium uptake with decreasing PVP film thickness were observed on the HMDS surfaces, while the SiO_x surface exhibited thickness independent equilibrium absorption. The decreased absorption with decreasing film thickness is attributed a depletion layer of water near the polymer/HMDS interface, arising from hydrophobic interactions between the surface and water. The diffusivity of water decreased when the film thickness was less than 60 nm, independent of the surface treatment. Changes in the properties of ultrathin polymer films occur even in plasticized films containing nearly 50% water.     

πSynthesis and evaluation of a series of α-hydroxy ethers derived from isopropyl oleate

Several fatty derivatives with bulky moieties were prepared by treatment of epoxidized isopropyl oleate with a number of alcohols in the presence of sulfuric acid catalyst to provide a series of α-hydroxy ethers in good yield. The materials were analyzed for cold flow performance through cloud point and pour point determinations. The most promising α-hydroxy ether produced in this study, with respect to both low temperature behavior and economic criteria, was isopropyl 9(10)-(2-ethylhexoxy) 10(9)-hydroxystearate, which has a cloud point of −23°C and pour point of −24°C.     

Determination of residual organic matter in extracted oil sands using a low temperature ashing method

The application of a low temperature ashing method for estimating total residual organic matter (toluene insolubles) in oil sands is described. A linear correlation exists between organic carbon content and loss on ignition at 400 ± 10 °C of solvent extracted oil sands. The ratio between total organic carbon and the weight loss on ignition (CT/LOl) owing to the removal of residual organic matter is much lower than that obtained for toluene soluble bitumen fractions, indicating very different chemical composition for the residual organic matter. The measured carbon content of the residual organic matter in oil sands suggests that this material could be a mixture of various fractions contained in resins, asphaltenes, asphaltic acids and humic acids.     

Radical styrene polymerization in the presence of trace levels of sulfonic acids

The bulk polymerization of styrene in the presence of the vinyl functional sulfonic acid 2‐sulfoethylmethacrylate (SEM) was found to have utility for making polystyrenes with narrow polydispersity, bimodal polydispersity, and ultrahigh molecular weight at fast polymerization rates. Narrow polydispersity polymers were made by the addition of SEM to nitroxide‐mediated polymerizations. Bimodal polydispersity polymers were made by the ultrahigh molecular weight component being made in the presence of SEM in the absence of an initiator and the low molecular weight component being made in the presence of an initiator and/or chain‐transfer agent. Ultrahigh molecular weight monomodal polystyrenes were prepared at much faster polymerization rates than possible via spontaneous polymerization in the absence of SEM. SEM was found to be more effective, by an order of magnitude, than camphor sulfonic acid on a weight basis and, because it is copolymerized into the polymer chain, should not lead to corrosion problems during fabrication of the polymer. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 87: 869–875, 2003