Structural and kinetic studies on ligand binding in wild-type and active-site mutants of penicillin acylase

Penicillin acylase catalyses the condensation of Calpha-substituted phenylacetic acids with beta-lactam nucleophiles, producing semi-synthetic beta-lactam antibiotics. For efficient synthesis a low affinity for phenylacetic acid and a high affinity for Calpha-substituted phenylacetic acid derivatives is desirable. We made three active site mutants, alphaF146Y, betaF24A and alphaF146Y/betaF24A, which all had a 2- to 10-fold higher affinity for Calpha-substituted compounds than wild-type enzyme. In addition, betaF24A had a 20-fold reduced affinity for phenylacetic acid. The molecular basis of the improved properties was investigated by X-ray crystallography. These studies showed that the higher affinity of alphaF146Y for (R)-alpha-methylphenylacetic acid can be explained by van der Waals interactions between alphaY146:OH and the Calpha-substituent. The betaF24A mutation causes an opening of the phenylacetic acid binding site. Only (R)-alpha-methylphenylacetic acid, but not phenylacetic acid, induces a conformation with the ligand tightly bound, explaining the weak binding of phenylacetic acid. A comparison of the betaF24A structure with other open conformations of penicillin acylase showed that betaF24 has a fixed position, whereas alphaF146 acts as a flexible lid on the binding site and reorients its position to achieve optimal substrate binding.     

Measurement of soybean fatty acids by near-infrared spectroscopy: Linear and nonlinear calibration methods

A key element of successful development of new soybean cultivars is availability of inexpensive and rapid methods for measurement of FA in seeds. Published research demonstrated applicability of NIR spectroscopy for FA profiling in oilseeds. The objectives of this study were to investigate the applicability of NIR spectroscopy for measurement of FA in whole soybeans and compare performance of calibration methods. Equations were developed using partial least squares (PLS), artificial neural networks (ANN), and support vector machines (SVM) regression methods. Validation results demonstrated that (i) equations for total saturates had the highest predictive ability (r ²=0.91–0.94) and were usable for quality assurance applications, (ii) palmitic acid models (r ²=0.80–0.84) were usable for certain research applications, and (iii) equations for stearic (r ²=0.49–0.68), oleic (r ²=0.76–0.81), linoleic (r ²=0.73–0.76), and linolenic (r ²=0.67–0.74) acids could be used for sample screening. The SVM models produced significantly more accurate predictions than those developed with PLS. ANN calibrations were not different from the other two methods. Reduction in the number of calibration samples reduced predictive ability of all equations. The rate of performance degradation of SVM models with sample reduction was the lowest.     

Nanoscratch testing to assess the fiber adhesion of short‐carbon‐fiber composites

In a composite material, the degree of adhesion between the fiber and the matrix plays an important role in the overall performance of the material. Because the load between the fiber and the matrix is realized throughout the interphase region material, a lot of effort has gone into characterizing the strength of the interphase. In this study, nanoscratch tests on the composite samples were used to provide a relative measure of adhesion in different composite materials. Carbon‐filled nylon 6,6 and polycarbonate resins were evaluated with this method. The carbon fillers we used were polyacrylonitrile‐based carbon fibers sized and surface‐treated for the respective matrix and pitch‐based carbon fibers without any sizing or surface treatment. Tensile and X‐ray photoelectron spectroscopy data for the composites we considered are also presented to compare to the nanoscratch results. It is shown that nanoscratch testing on the composites, with the proposed data analysis, can be an effective tool for determining the relative degree of adhesion between different composites. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 328–335, 2007     

Preparation and characterization of PLLA/P(CL-b-LLA) blends by an in situ ring-opening polymerization

This paper describes a new reactive blending approach to improve the compatibility of poly(l-lactide) (PLLA) and poly(?-caprolacton) (PCL). For this purpose, the ring-opening polymerization of l-lactide (LLA) was carried out in the presence of PCL-OH (OH groups on one end). The P(CL-b-LLA) block copolymers were in situ formed during polymerization, and as a result, PLLA/P(CL-b-LLA) blends were obtained. The characterization of GPC and ¹H NMR verified the synthesis of PLLA/P(CL-b-LLA) blends. Furthermore, the ¹³C NMR spectroscopy showed that no transesterification reaction occurred to a significant extent during LLA polymerization process. The in situ formed P(CL-b-LLA) compatibilizes the phase separated structure of PCL domains in PLLA matrix. The size of PCL domains in PLLA matrix became much smaller than that in a solution blended sample. The average sizes of PCL domains are controllable in the level of sub-micron scale.     

Computationally efficient solution of population balance models incorporating nucleation, growth and coagulation: application to emulsion polymerization

A computationally efficient solution technique is presented for population balance models accounting for nucleation, growth and coagulation (aggregation) (with extensions for breakage). In contrast to earlier techniques, this technique is not based on approximating the population balance equation, but is based on employing individual rates of nucleation, growth and coagulation to update the PSD in a hierarchical framework. The method is comprised of two steps. The first step is the calculation of the rates of nucleation, growth and coagulation by solving an appropriate system of equations. This information is then used in the second step to update the PSD. The method effectively decomposes the fast and the slow kinetics, thereby eliminating the stiffness in the solution. In solving the coagulation kernel, a semi-analytical solution strategy is adapted, which substantially reduces the computational requirement, but also ensures the consistency of properties such as the number and mass of particles.     

Supramolecular complexes: lamellar structure and crystalline transformation

Interfacial reaction of highly-branched polyethyleneimine (PEI) with octadecanoic acid (OA) was performed to prepare a series of supramolecular complexes (PEI(OA)_x). The complexes at solid state have typical lamellar structure, and the interlamellar distance can be modulated by the OA content. The long period values of the supramolecular complexes measured by SAXS were found to be dependent on the compositions, which are in good consistency with those measured by TEM. For example, the long period values from SAXS for PEI(OA)_0.76, PEI(OA)_1.03, and PEI(OA)_1.67 were 46.5, 62.7, and 56.2 Å, respectively. The corresponding data from TEM were 45.7, 60.7, and 56.6 Å, respectively. A model was proposed for the construction mode of the side alkyl chains (crystallization region) associated with PEI backbone (amorphous region), in which the side alkyl chains were arranged to be ‘end-to-end’ packing for the x=1 complex, while an ‘interdigitated structure’ of the side alkyl chains was deduced for the x>1 and x<1 complexes. Temperature variable FT-IR combination investigation of the scissoring band, rocking band, and stretching band of methylene (CH₂) and vibrational band of carbonyl group (CO) indicated that the crystalline form of the crystallization region in the lamellae can be transformed from orthorhombic to hexagonal with the temperature increasing, and vice versa.     

Synthesis of nanodispersed oxides of vanadium, titanium, molybdenum, and tungsten on mesoporous silica using atomic layer deposition

The advantages of the atomic layer deposition (ALD) method for preparation of tungsten, vanadium, titanium, and molybdenum oxide catalyst supported on mesoporous silica are discussed, with emphasis on the importance of synthesis conditions on dispersion, structure and activity of the resulting materials. A suite of complementary techniques such as DRS-UV/Vis, BET, ¹H-NMR, XRD, and TEM were used to study the structural properties of the supported metal oxides, and probe reactions such as 2-butanol dehydration and ethanol partial oxidation were used to demonstrate the potential advantages of the ALD-prepared catalysts. Specifically, highly dispersed oxides of titanium, molybdenum, and tungsten oxide on mesoporous silica were synthesized using the ALD method. It is also demonstrated that attainment of high dispersions of vanadium oxide on mesoporous silica requires the presence of at least a single layer of titanium oxide due to the well-known poor interaction between vanadia and silica. The highly dispersed catalysts prepared here by ALD methods exhibited superior catalytic performance relative to those prepared using conventional incipient wetness impregnation.     

Polybutadiene‐modified clay and its polystyrene nanocomposites

Vinylbenzyl chloride was allowed to react with a low‐molecular‐weight oligomer of butadiene in order to give a material that could then be used to alkylate an amine to form an ammonium salt. This ammonium salt was ion‐exchanged onto a clay to give a new organically modified clay, which was used to prepare polystyrene‐clay nanocomposites by solution and bulk polymerization and by melt blending. The nanocomposites were characterized by using X‐ray diffraction, transmission electron microscopy, thermogravimetric analysis, cone calorimetry, and the evaluation of mechanical properties. These systems show good nanodispersion and improvement in thermal and mechanical properties. J. Vinyl Addit. Technol. 10:44–51, 2004. © 2004 Society of Plastics Engineers.     

Functionally substituted tetramethylcyclopentadienes: Synthesis of 1-(p-hydroxyphenyl)-2,3,4,5-tetramethylcyclopentadiene and its use in the synthesis of an organometallic polymer

Reaction of 2,3,4,5-tetramethylcyclopent-2-enone (1) withp-LiC₆H₄OC-Me₂OMe, followed by treatment with aqueous acid afforded 1-(p-hydroxyphenyl)-2,3,4,5-tetramethylcyclopentadiene (2). This new ligand was then used in the synthesis of the functionally substituted organomolybdenum reagent (⁵-C₅Me₄-p-C₆H₅OH)Mo(CO)₂(NO) (3). Treatment of a preformed 1/1 styrene/maleic anhydride copolymer ( ) with 5 mol% 3 led to chemical incorporation of the organometallic species into the polymer. The final product contained 3 mol% of the organometallic moiety.