The influence of pretreatment and enzyme loading on the effectiveness of batch and fed-batch hydrolysis of corn stover.

To try to improve hydrolysis yields at elevated solids loadings, a comparison was made between batch and fed-batch addition of fresh substrate at the initial and later phases of hydrolysis. Both ethanol (EPCS) and steam-pretreated corn stover (SPCS) substrates were tested at low (5 FPU) and high (60 FPU) loadings of cellulase per gram of cellulose. The fed-batch addition of fresh substrate resulted in a slight decrease in hydrolysis yields when compared with the corresponding batch reactions. A 72-h hydrolysis of the SPCS substrate resulted in a hydrolysis yield of 66% compared with 51% for the EPCS substrate. When the enzyme adsorption and substrate characteristics were assessed during batch and fed-batch hydrolysis, it appeared that the irreversible binding of cellulases to the more recalcitrant original substrate limited their access to the freshly added substrate. After 72-h hydrolysis of the SPCS substrate at low enzyme loadings, ∼40-50% of the added cellulases were desorbed into solution, whereas only 20% of the added enzyme was released from the EPCS substrate. Both simultaneous and sequential treatments with xylanases and cellulases resulted in an up to a 20% increase in hydrolysis yields for both substrates at low enzyme loading. Simons' stain measurements indicated that xylanase treatment increased cellulose access, thus facilitating cellulose hydrolysis.

Xyloglucan endo-transglycosylase-mediated xyloglucan rearrangements in developing wood of hybrid aspen.

Xyloglucan endo-transglycosylases (XETs) encoded by xyloglucan endo-transglycosylases/hydrolase (XTH) genes modify the xyloglucan-cellulose framework of plant cell walls, thereby regulating their expansion and strength. To evaluate the importance of XET in wood development, we studied xyloglucan dynamics and XTH gene expression in developing wood and modified XET activity in hybrid aspen (Populus tremula × tremuloides) by overexpressing PtxtXET16-34. We show that developmental modifications during xylem differentiation include changes from loosely to tightly bound forms of xyloglucan and increases in the abundance of fucosylated xyloglucan epitope recognized by the CCRC-M1 antibody. We found that at least 16 Populus XTH genes, all likely encoding XETs, are expressed in developing wood. Five genes were highly and ubiquitously expressed, whereas PtxtXET16-34 was expressed more weakly but specifically in developing wood. Transgenic up-regulation of XET activity induced changes in cell wall xyloglucan, but its effects were dependent on developmental stage. For instance, XET overexpression increased abundance of the CCRC-M1 epitope in cambial cells and xylem cells in early stages of differentiation but not in mature xylem. Correspondingly, an increase in tightly bound xyloglucan content was observed in primary-walled xylem but a decrease was seen in secondary-walled xylem. Thus, in young xylem cells, XET activity limits xyloglucan incorporation into the tightly bound wall network but removes it from cell walls in older cells. XET overexpression promoted vessel element growth but not fiber expansion. We suggest that the amount of nascent xyloglucan relative to XET is an important determinant of whether XET strengthens or loosens the cell wall.

Extraction of water-soluble hemicelluloses from barley husks.

Heat treatment of barley husks was performed to extract arabinoxylan with high yield and high weight-averaged molecular mass (M(W)). Microwave irradiation was employed for initial screening of suitable residence times (2-15 min), temperatures (120-210 degrees C) and initial pH (3-13) of the reaction slurry. Microwave irradiation was shown to be a good method for predicting the effects of heat treatment on a larger scale using steam pretreatment. A M(W) of about 40,000 Da was achieved without the addition of chemicals, by both microwave irradiation and steam pretreatment, with a yield of about 9%. The yield was significantly increased by slightly increasing the severity factor. However, the M(W) decreased below 20,000 Da at severity factors above 3.7. Arabinosyl side groups were enzymatically hydrolysed from the arabinoxylan by alpha-L-arabinofuranosidase to a degree of 47%, demonstrating the ability to specifically alter the side group substitution of arabinoxylans with the use of enzymes.

Protein release from galactoglucomannan hydrogels: influence of substitutions and enzymatic hydrolysis by beta-mannanase.

O-Acetyl-galactoglucomannan (AcGGM) is the major soft-wood hemicellulose. Structurally modified AcGGM and hydrogels of AcGGM were prepared. The degree of substitution (DS) of AcGGM was modified enzymatically with alpha-galactosidase, and chemically with an acrylate derivative, 2-hydroxyethylmethacrylate (HEMA). The hydrolysis of AcGGM with beta-mannanase was shown to increase with decreasing DS. AcGGM hydrogels were prepared from chemically modified AcGGM with varying DS of HEMA. Bovine serum albumin (BSA) was encapsulated in hydrogels. A spontaneous burst release of BSA was decreased with increased DS of HEMA. The addition of beta-mannanase significantly enhanced the BSA release from hydrogels with a DS of 0.36, reaching a maximum of 95% released BSA after eight hours compared to 60% without enzyme. Thus, both the pendant group composition and the enzyme action are valuable tools in the tailoring of hydrogel release profiles of potential interest for intestine drug delivery.

Assessing the effect of latex particle size and distribution on the rheological and adhesive properties of model waterborne acrylic pressure-sensitive adhesives films.

The adhesive and rheological properties of model acrylic pressure-sensitive adhesive (PSA) films prepared from high solid emulsions with different particle sizes and distributions have been investigated with a customized probe tack apparatus. For each emulsion, the monomer composition and gel content were kept constant but different average particle sizes and distributions were used. Adhesive films 100 microm thick were then prepared from these emulsions and their rheological properties in the linear regime and adhesive properties were systematically characterized. Surprisingly, both the rheological and adhesive properties were found to be very dependent on the initial latex particle size distribution. A series of experiments were carried out to assess the adhesive properties of films made from blends of small- and large-particle-size latexes. Using the probe tack test, a maximum in adhesion energy of the dry films was found for 60% of small particles in the blend, a composition clearly different from that giving a minimum viscosity of the latex implying that optimizing for properties may not be equivalent to optimizing for processing in these adhesive applications. Finally, the adhesive properties of two multimodal latexes with different particle size distributions were investigated. Both gave significantly higher adhesion energies and clear evidence of a fibrillar detachment process. This important result suggests that the spatial distribution of gel domains in the dry film and the molecular connectivity between those gel domains also play an important role in controlling its adhesive properties.

Effects of geometric confinement on the adhesive debonding of soft elastic solids.

The effect of increasing confinement on soft elastic gel layers has been investigated and a means of analyzing the behavior of such systems has been developed. A probe tack test was used to study the behavior of thin elastic layers during interfacial debonding from a cylindrical glass indenter. For this gel-indenter system, confinement is defined as the ratio of a(0), the radius of the indenter, to h, the thickness of the elastic layer. In order to investigate geometric effects, the adhesion energy of the gel was kept constant while the thickness and modulus of the gels were varied. A fracture mechanics approach, based on the compliance of the layer, has been employed in analyzing the experimental data. It is shown that a fracture mechanics analysis is appropriate for these systems, allowing quantitative results to be obtained, despite very irregular contacts. It has also been shown that the interfacial instabilities observed during debonding maximize the compliance of the elastic layer. Additionally, four dimensionless parameters that dictate the behavior of confined systems have been defined, offering a general guide to understanding and characterizing the mechanical behavior of thin elastic layers.