Interactions between immunoglobulin-like and catalytic modules in Clostridium thermocellum cellulosomal cellobiohydrolase CbhA

Cellobiohydrolase CbhA from Clostridium thermocellum cellulosome is a multi-modular protein composed starting from the N-terminus of a carbohydrate-binding module (CBM) of family 4, an immunoglobulin(Ig)-like module, a catalytic module of family 9 glycoside hydrolases (GH9), X1(1) and X1(2) modules, a CBM of family 3 and a dockerin module. Deletion of the Ig-like module from the Ig-GH9 construct results in complete inactivation of the GH9 module. The crystal structure of the Ig-GH9 module pair reveals the existence of an extensive module interface composed of over 40 amino acid residues of both modules and maintained through a large number of hydrophilic and hydrophobic interactions. To investigate the importance of these interactions between the two modules, we compared the secondary and tertiary structures and thermostabilities of the individual Ig-like and GH9 modules and the Ig-GH9 module pair using both circular dichroism (CD) spectroscopy and differential scanning calorimetry (DSC). Thr230, Asp262 and Asp264 of the Ig-like module are located in the module interface of the Ig-GH9 module pair and are suggested to be important in 'communication' between the modules. These residues were mutated to alanyl residues. The structure, stability and catalytic properties of the native Ig-GH9 and its D264A and T230A/D262A mutants were compared. The results indicate that despite being able to fold relatively independently, the Ig-like and GH9 modules interact and these interactions affect the final fold and stability of each module. Mutations of one or two amino acid residues lead to destabilization and change of the mechanism of thermal unfolding of the polypeptides. The enzymatic properties of native Ig-GH9, D264A and T230A/D262A mutants are similar. The results indicate that inactivation of the GH9 module occurs as a result of multiple structural disturbances finally affecting the topology of the catalytic center.     

Quantitative probing the interfacial structure of TPO/CPO blends by transmission electron microscopy via EDX

High-resolution scanning transmission electron microscope (HR STEM) measurements were performed on a thermoplastic polyolefin (TPO) substrate coated with chlorinated polyolefin (CPO). This CPO was a maleated chlorinated polypropylene containing 21.8 wt% Cl. The TPO investigated was a blend of high-modulus isotactic polypropylene (iPP) with a crystalline ethylene-butene copolymer (EB9) containing 9 wt% butene. For these injection-molded samples, examined ca. 10 mm from the mold gate, a stratified morphology found at TPO surface consisting of thin fibers of EB9 trapped in a transcrystalline iPP matrix, with crystalline lamellae propagating from the matrix across the EB9 domains. This structure was unperturbed when the plaques were coated (from tetrahydrofuran solution) with a 5 μm layer of CPO, but underwent changes of increasing severity when subjected either to a dry bake at 120 °C or annealing at 120 °C in the presence of xylene vapor. The interfacial structure between the CPO and the TPO was probed by TEM with energy dispersive X-ray imaging (EDX). The elemental chlorine across the interface gave good fits to a tan h function, and the interface thickness increased from 23 ± 2 nm to 28 ± 1 nm upon annealing at 120 °C for 30 min. After annealing in the presence of xylene vapor, this value increased to 50 ± 4 nm. As reference points, we determined an interface thickness of 29 ± 3 nm for the CPO-EB interface and 15 ± 2 nm for the interface between CPO and iPP.     

Enzymatic transesterification for production of biodiesel using yeast lipases: An overview

Biodiesel has provided an eco-friendly solution to fuel crisis, as it is renewable, biodegradable and a non-toxic fuel that can be easily produced through enzymatic transesterification of vegetable oils and animal fats. Enzymatic production of biodiesel has many advantages over the conventional methods as high yields can be obtained at low reaction temperatures with easy recovery of glycerol. Microbial lipases are powerful biocatalysts for industrial applications including biodiesel production at lower costs due to its potential in hydrolyzing waste industrial materials. Among them, lipases from yeasts, Candida antarctica, Candida rugosa, Cryptococcus sp., Trichosporon asahii and Yarrowia lipolytica are known to catalyze such reactions. Moreover, stepwise addition of methanol in a three step, two step and single step reactions have been developed using yeast lipases to minimize the inhibitory effects of methanol. The latest trend in biodiesel production is the use of whole-cell as biocatalysts, since the process requires no downstream processing of the enzyme. Synthesis of value added products from the byproduct glycerol further reduces the production cost of biodiesel. This review aims at compiling the information on various yeast lipase catalyzed transesterification reactions for greener production of biodiesel.     

Structure/property relationships in flexible alkoxysilane automotive coatings

Flexible automotive coatings are susceptible to scratch and mar damage, especially during finishing and assembly operations. One-component (1K) flexible clearcoats exhibit very good scratch and mar resistance, but unfortunately suffer from poor durability and environmental etch resistance. Two-component clearcoats offer improvements in both etch and durability, but at the expense of scratch and mar. In this paper, the concept and properties of 1K flexibilized silane clearcoats for use on automotive plastics will be introduced and their structure/property relationships examined as they apply to scratch and mar. The role of coating crosslink density, toughness, glass transition temperature (T_g), and surface profile on the scratch damage of coated plastic substrates will be described. In addition, a new scratch methodology, termed Scratcho, is utilized to determine relative scratch performance and is compared to conventional scratch resistance testing. Results to date indicate that hardness, as affected by the glass transition temperature, and crosslink density, as it contributes to higher essential work values, both affect resultant scratch propensity of the flexible coatings. The relative ranking of different coating systems employing alternate crosslinkers (e.g., isocyanate and melamine) is also presented and compared to the newly developed silane crosslinked coatings. Presented at the 28th International Waterborne, High-Soids, and Powder Coatings Symposium, Feb. 21–23, 2001, in New Orleans. LA. 377 Fairall St., Ajax, Ont., L1S 1R7, Canada. 401 Southfield Rd., P.O. Box 6231, Dearborn, MI 48121-6231.     

Adhesion of CPO onto high modulus TPO: Lap-shear tests in conjunction with microscopy studies of the fracture surface structure

A lap-shear test was employed to investigate the failure mechanism of a chlorinated polyolefin (CPO) coating on a high-modulus thermoplastic olefin (TPO) substrate fabricated as a blend of a highly crystalline Ziegler-Natta isotactic polypropylene (iPP) and a crystalline metallocene poly(ethylene-butene) (9 wt% butene, EB9) impact modifier. The CPO was a chlorinated polypropylene containing 20 wt% Cl. The results showed that the fracture strength increased with increasing EB9 content in TPO blends. They also showed that the presence of xylene vapor during the bake step improved the adhesion between CPO and iPP itself (by 40%), but had a much smaller effect for the TPOs. Optical and transmission electronic microscopy images revealed a well-defined skin layer approximately 230 μm thick at the mold surface of the injection molded substrates. For the 25 wt% EB9 blend (TPO₂₅), this skin layer consists of thin fibers of EB trapped in a transcrystalline iPP matrix, with crystalline lamellae propagating from the matrix across the EB9 domains. Laser scanning confocal fluorescence microscopy (LCFM) and scanning electron microscopy images of iPP/CPO/iPP samples indicate that failure occurred close to the interface between the CPO and the iPP substrate, and, during fracture, the CPO layer maintained its original thickness. For the TPO/CPO/TPO sandwich samples, the fracture surfaces themselves were much rougher than that between CPO and iPP. Substantial deformation of the CPO layer was seen in the fractured samples, and failure was due primarily to cohesive fracture of the CPO in the region adjacent to the TPO substrate. From the perspective of newly introduced environmental regulations restricting aromatic hydrocarbons in automotive coatings, the most important result was the strong adhesion between CPO and TPO₂₅, with little difference between the samples exposed to xylene vapor and those not exposed to xylene.     

Testing a mathematical model for initial chemical reaction fouling using a dilute protein solution

A 1 wt% lysozyme solution was used as a model fluid to test a previously formulated mathematical model for the initial chemical reaction fouling rate of a heat transfer surface. The experimental results showed that, at a given wall temperature, a maximum initial fouling rate existed over a range of fluid velocities. The maximum rate and the fluid velocity at which it occurred both increased as the wall temperature increased. These observations were consistent with the model. Quantitatively, the average absolute percent deviation between the experimental results and the optimum model predictions was 23,3%. The decrease in initial fouling rate with increasing velocity at high fluid velocities was even greater than predicted by the model.     

Dung pads increase pasture production,soil nutrients and microbial biomass carbon in grazed dairy systems

In grazing systems dung is an important source of nutrients which can increase soil fertility and contribute to nutrient cycling through increased pasture production. Changes in soil chemical and biological properties and pasture production were measured below and around dung pads created in the field. Almost 65% of the total dung P remained after 45 days and about two-thirds of the pad fresh weight had disappeared in that time, indicating that physical degradation is the mechanism of incorporation of dung P. All the pads bar one were completely degraded by 112 days. At this time, soil pH and EC increased under dung pads as did Olsen extractable inorganic phosphorus (Pi) and total phosphorus (Pt), with these changes observed at 0–5 and 5–10 cm depths. Bicarbonate extractable soil organic phosphorus (Po) was not affected by dung and the observed differences in soil Po:Pi ratios were largely influenced by the substantial addition of inorganic P from dung. Dung increased the P buffering capacity of the 0–5 cm soil samples collected at the end of the experiment, potentially contributing to the increased extractable soil P measured under the pads. Dung also changed soil properties in 0–10 cm samples with increases in soil pH, EC, Colwell P and Colwell K recorded even long after the dung had completely disappeared. Microbial biomass carbon increased under dung pads in the 0–10 cm soil samples in the first 45 days after pads were applied. Total herbage production and ryegrass biomass increased significantly under and around the pads by 112 days after dung was applied. The botanical composition changed significantly with increased ryegrass contents observed, but only under the dung pads. This experiment demonstrated that increases in pasture around dung pads in the field are not solely due to animal rejection.     

2003 Roy W. Tess Award

Conclusions Mechanical interlocking of topcoat with the nonpolar TPO surface can be achieved through the use of an adhesion promoter, namely a chlorinated poly(olefin). The type of CPO used, in addition to the types of solvents and heat effects used, can substantially influence the degree of adhesion/cohesion obtained within the CPO.TPO system. Heat histories, TPO molding variations, CPO types, including solvent and resin variations, and topcoat (basecoat/clearcoat) chemistries were all found to influence the adhesion/cohesion of the painted TPO assembly. Surface damage resistance was found to mirror the effects of adhesion as described earlier. Control of the interphase formed between the TPO substrate and the subsequent topcoat layers becomes increasingly important if one wishes to maintain damage resistance within the painted composite. Testing methodology development, namely “gouge” chip, abrasion, and scratch resistance, is paramount in predicting performance under specified loads. Through interpretation of data received in the various testing methodologies, the mechanical properties of the topcoat/substrate combination may be varied to obtain the performance required in a variety of applications. The Roy W. Tess Award in Coatings is presented annually by the Division of Polymeric Materials: Science and Engineering (PMSE) in recognition of outstanding contributions to coatings science and technology. Funded by a grant to the Division by Dr. and Mrs. Roy W. Tess, the purpose of the award is to encourage interest and progress in coatings and recognize significant contributions to the field. Dr. Rose Ryntz, Manager and Staff Technical Fellow with Visteon Corporatiom, Dearborn, MI, received the award from Dr. Paul Valint, Jr., Chair of the PMSE Division in September 2003 during the 226th meeting of the American Chemical Society in New York, NY. Dr. Ryntz's award address followed the Award Symposium. The following papers were presented at that symposium.     

Pt-V2O5-WO3/TiO2 catalysts supported on SiC filter for NO reduction at low temperature

The catalytic filter, V₂O₅-WO₃-TiO₂ supported on a ceramic filter, is known as a promising material for treating particulates and NO _x simultaneously at optimum temperatures around 320°C. In order to improve its catalytic activity at low temperatures, the effect of Pt addition on the catalytic filter has been investigated. Catalytic filters, Pt-V₂O₅-WO₃-TiO₂/SiC, were prepared by co-impregnation of Pt, V, and W precursors on TiO₂ coated-SiC filter by vacuum aided-dip coating. The Pt-added catalytic filter shifted the optimum working temperature from 280–330°C (for the non Pt-impregnated filter) to 180–230°C, providing N _x slip concentration less than 20 ppm for the treatment of 700 ppm NO at a face velocity of 2 cm/s with the same value over the non Pt-added catalytic filters. The promotional effect following the addition of Pt is believed to result from electrical modification of the catalyst maintaining a high electron transfer state. Ammonia oxidation was also observed to be dominant above the optimal temperature for SCR.     

High surface area carbide-derived carbon fibers produced by electrospinning of polycarbosilane precursors

Highly porous carbide-derived carbon fibers have been synthesized by electrospinning of polycarbosilane with subsequent pyrolysis and chlorination. The resulting ultrathin fibers show specific surface areas up to 3116 m² g⁻¹ and very high storage capacities for hydrogen up to 3.86 wt.% at 17 bar and 77 K. Due to the outstanding adsorption performance and other properties such as high temperature stability and the unique CDC fiber shape, this new kind of fiber material offers promising possibilities for several applications like air or liquid filters or textiles for protective clothing. Application as a flexible electrode material for supercapacitors is conceivable.