3- Instead of 4-helix formation in a de novo designed protein in solution revealed by small-angle X-ray scattering

De novo design and chemical synthesis of proteins and their mimics are central approaches for understanding protein folding and accessing proteins with novel functions. We have previously described carbohydrates as templates for the assembly of artificial proteins, so-called carboproteins. Here, we describe the preparation and structural studies of three alpha-helical bundle carboproteins, which were assembled from three different carbohydrate templates and one amphiphilic hexadecapeptide sequence. This heptad repeat peptide sequence has been reported to lead to 4-alpha-helix formation. The low resolution solution structures of the three carboproteins were analyzed by means of small-angle X-ray scattering (SAXS) and synchrotron radiation circular dichroism (SRCD). The ab initio SAXS data analysis revealed that all three carboproteins adopted an unexpected 3+1-helix folding topology in solution, while the free peptide formed a 3-helix bundle. This finding is consistent with the calculated alpha-helicities based on the SRCD data, which are 72 and 68 % for two of the carboproteins. The choice of template did not affect the overall folding topology (that is for the 3+1 helix bundle) the template did have a noticeable impact on the solution structure. This was particularly evident when comparing 4-helix carboprotein monomers with the 2x2-helix carboprotein dimer as the latter adopted a more compact conformation. Furthermore, the clear conformational differences observed between the two 4-helix (3+1) carboproteins based on D-altropyranoside and D-galactopyranoside support the notion that folding is affected by the template, and subtle variations in template distance-geometry design may be exploited to control the solution fold. In addition, the SRCD data show that template assembly significantly increases thermostability.     

Controlled release of biocide from silica microparticles in wood paint

Porous microparticles represent an attractive encapsulation platform as they provide a feasible route to a wide range of encapsulated chemical compounds. This assures an increased lifetime of the encapsulated compound by protecting it from the surrounding environment as well as providing a way to control the release rate. Porous microparticles encapsulating the commonly used biocide for wood protection, 3-iodoprop-2-ynyl N-butylcarbamate (IPBC), have been synthesized. The microparticles are spherical with an average size distribution of roughly 1 μm as determined by scanning electron microscopy (SEM). Determination of the release rate of the biocide from microcapsules incorporated into the paint film has been performed under diluted conditions. These experiments show that a slower release of the biocide can be obtained by encapsulation. Additionally, the microparticles have been shown to prolong the biocidal effect of the fungicide under accelerated weathering tests by protecting the biocide from UV-induced chemical degradation.     

Stochastic response of hysteretic systems

This paper deals with approximate stochastic response of hyseretic structures under white noise excitation, based on It stochastic differential equations. Instead of the original system an equivalent nonlinear system is considered, in which the drift vector is given by a series expansion of the order n 1 in terms of the state variables. n = 1 represents the well-known case of equivalent linearization. Components in the drift vector representing the nonanalytical constitutive equations are replaced by a cubic polynomial expansion. The hierarchy of statistical moment equations is closed by a cumulant neglect closure scheme. The method has been applied to a bilinear single degree-of-freedom system subjected to white noise excitation, for which an equivalent system with a cubic series expansion to the constitutive equation is considered. The results obtained are compared with those of numerical simulation and alternative methods, and they provide substantial improvements compared to equivalent linearization.     

Degradation of phthalate esters in an activated sludge wastewater treatment plant

Efficient removal of phthalate esters (PE) in wastewater treatment plants (WWTP) is becoming an increasing priority in many countries. In this study, we examined the fate of dimethyl phthalate (DMP), dibutyl phthalate (DBP), butylbenzyl phthalate (BBP), and di-(2-ethylhexyl) phthalate (DEHP) in a full scale activated sludge WWTP with biological removal of nitrogen and phosphorus. The mean concentrations of DMP, DBP, BBP, and DEHP at the WWTP inlet were 1.9, 20.5, 37.9, and 71.9 μg/L, respectively. Less than 0.1%, 42%, 35%, and 96% of DMP, DBP, BBP, and DEHP was associated with suspended solids, respectively. The overall microbial degradation of DMP, DBP, BBP, and DEHP in the WWTP was estimated to be 93%, 91%, 90%, and 81%, respectively. Seven to nine percent of the incoming PE were recovered in the WWTP effluent. Factors affecting microbial degradation of DEHP in activated sludge were studied using [U-¹⁴C-ring] DEHP as tracer. First order rate coefficients for aerobic DEHP degradation were 1.0×10⁻², 1.4×10⁻², and 1.3×10⁻³ at 20, 32, and 43 °C, respectively. Aerobic degradation rates decreased dramatically under aerobic thermophilic conditions (<0.1×10⁻² h⁻¹ at 60 °C). The degradation rate under anoxic denitrifying conditions was 0.3×10⁻² h⁻¹, whereas the rate under alternating conditions (aerobic-anoxic) was 0.8×10⁻² h⁻¹. Aerobic DEHP degradation in activated sludge samples was stimulated 5-9 times by addition of a phthalate degrading bacterium. The phthalate degrading bacterium was isolated from activated sludge, and maintained a capacity for DEHP degradation while growing on vegetable oil. Collectively, the results of the study identified several controls of microbial PE degradation in activated sludge. These controls may be considered to enhance PE degradation in activated sludge WWTP with biological removal of nitrogen and phosphorus.     

Characterization of activated sludge flocs by confocal laser scanning microscopy and image analysis

In this study we present a new approach to determine volumes, heterogeneity factors, and compositions of the bacterial population of activated sludge flocs by 3D confocal imaging. After staining the fresh flocs with fluorescein-isothiocyanate, 75 stacks of images (containing approx. 3000 flocs) were acquired with a confocal laser scanning microscope. The self-developed macro 3D volume and surface determination for the KS 400 software package combined the images of one stack to a 3D image and calculated the real floc volume and surface. We determined heterogeneity factors like the ratio of real floc surface to the surface of a sphere with the respective volume and the fractal dimension (D(f)). According to their significant influence on floc integrity and quality, we also investigated the chemical composition of flocs and quantified their bacterial population structure by using group-specific rRNA-targeted probes for fluorescence in situ hybridization. By a settling experiment we enriched flocs with poor settling properties and determined the above-mentioned parameters. This approach revealed shifts in floc volume, heterogeneity, and bacterial and chemical composition according to the settling quality of the flocs.     

A digital system for surface roughness analysis of plane and cylindrical parts

This paper describes an on-line system for digital analysis of surfaces where the electrical signal from a surface roughness instrument is converted and input into a desk-top computer which also controls the traversing. Special fixtures integrated with the system and dedicated software permit two- and three-dimensional tracings to be carried out on plane as well as on cylindrical parts. The surface profiles can be plotted in different ways. Roughness parameters, frequency spectra etc, can be computed digitally and all results can be stored. Scratches and irregularities on the surface can be identified and processed separately from the rest of the surface. The desk-top computer is connected to the university mainframe computer through a modem, this being of advantage in connection with graphic data processing. The capability of the equipment has been investigated and results from calibration are presented.     

A New Immobilized Glucose Isomerase with High Productivity Produced by a Strain of Streptomyces murinus

The properties of a new immobilized glucose isomerase, produced by a selected strain of Streptomyces murinus, are described. The major advantages of the new immobilized enzyme are a productivity of more than 10,000 kg syrup dry substance per kg enzyme under optimal industrial conditions, increased activity and a very low syrup by-product formation. The influence of process parameters (temperature, pH and feed syrup additives) on activity and stability is discussed based on laboratory and industrial plant data.     

Liquefaction,Saccharification and Isomerization of Starches from Sources Other than Maize

Examination of the suitability of a number of starches for the production of fructose syrup is described. In addition to maize starch, starch from wheat, potatoes and tapioca has been examined. Liquefaction, saccharification and purification were performed in pilot scale, whereas the produced syrups were isomerized in laboratory scale. The liquefactions were performed with Termamyl® in a jet cooker process. Saccharification with Amyloglucosidase Novo 150 L under standard conditions gave good DX values. After purification the syrups were isomerized in laboratory columns with Sweetzyme®. The results - activity and productivity of Sweetzyme - were for all syrups as good as or better than those obtained with dextrose solutions.     

Identification of microstructural mechanisms during densification of a TiAl alloy by spark plasma sintering

This work aims at identifying, by coupled scanning and transmission electron microscopy (SEM and TEM) observations, the densification mechanisms occurring when an atomized Ti-47Al-1W-1Re-0.2Si powder is densified by spark plasma sintering (SPS). For this purpose, interruptions of the SPS cycle have been performed to follow the evolution of the microstructure step by step. The powder particles exhibit a classical dendritic microstructure containing a large amount of out-of-equilibrium α phase. During heating-up, the microstructure undergoes successive transformations. At T = 525-875 °C the α phase transforms into γ. The γ phase formed is supersaturated in W and Re. It de-saturates for T above 875 °C by discontinuous precipitation of W and Re-rich B2 phase. Densification takes place for T between 900 °C and 1150 °C by plastic deformation of the powder particles. TEM observations show that the repartition of the plastic deformation is correlated to the dendritic microstructure, and that dynamic recrystallization mechanisms occur. Microstructural phenomena directly resulting from the high currents involved in the SPS process have not been observed.