. 2010. Effect of moisture on electrospun nanofiber composites of poly(vinyl alcohol) and cellulose nanocrystals. Biomacromolecules, volume 11, number 9, pages 2471-2477. © 2010 American Chemical Society (ACS)Reprinted with permission from American Chemical Society. Effect of Moisture on Electrospun Nanofiber Composites ofPoly(vinyl alcohol) and Cellulose Nanocrystals The effect of humidity on the morphological and thermomechanical properties of electrospun poly(vinyl alcohol) (PVA) fiber mats reinforced with cellulose nanocrystals (CNs) was investigated. Scanning electron microscopy (SEM) images revealed that the incorporation of CNs improved the morphological stability of the composite fibers even in high humidity environments. Thermal and mechanical properties of the electrospun fiber mats were studied by using differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), and large deformation tensile tests under controlled humidity and temperatures. The balance between the moisture-induced plasticization and the reinforcing effect of rigid CN particles was critical in determining the thermomechanical behaviors of the electrospun fiber mats. Results indicated that the stabilizing effect of the CNs in the PVA matrix might be compromised by water absorption, disrupting the hydrogen bonding within the structure. The amount of this disruption depended on the surrounding humidity and the CN loading. The reduction in tensile strength of neat PVA fiber mats as they were conditioned from low relative humidity (10% RH) to high relative humidity (70% RH) was found to be about 80%, from 1.5 to 0.4 MPa. When the structure was reinforced with CNs, the reduction in strength was limited to 40%, from 2 to 0.8 MPa over the same range in relative humidity. More importantly, the CN-loaded PVA fiber mats showed a reversible recovery in mechanical strength after cycling the relative humidity. Finally, humidity treatments of the composite PVA fiber mats induced significant enhancement of their strength as a result of the adhesion between the continuous matrix and the CNs.

BackgroundThe oxidation of carbohydrates from lignocellulose can facilitate the synthesis of new biopolymers and biochemicals, and also reduce sugar metabolism by lignocellulolytic microorganisms, reserving aldonates for fermentation to biofuels. Although oxidoreductases that oxidize cellulosic hydrolysates have been well characterized, none have been reported to oxidize substituted or branched xylo-oligosaccharides. Moreover, this is the first report that identifies amino acid substitutions leading to GOOX variants with reduced substrate inhibition.ResultsThe recombinant wild type gluco-oligosaccharide oxidase (GOOX) from the fungus Sarocladium strictum, along with variants that were generated by site-directed mutagenesis, retained the FAD cofactor, and showed high activity on cello-oligosaccharide and xylo-oligosaccharides, including substituted and branched xylo-oligosaccharides. Mass spectrometric analyses confirmed that GOOX introduces one oxygen atom to oxidized products, and 1H NMR and tandem mass spectrometry analysis confirmed that oxidation was restricted to the anomeric carbon. The A38V mutation, which is close to a predicted divalent ion-binding site in the FAD-binding domain of GOOX but 30 Å away from the active site, significantly increased the kcat and catalytic efficiency of the enzyme on all oligosaccharides. Eight amino acid substitutions were separately introduced to the substrate-binding domain of GOOX-VN (at positions Y72, E247, W351, Q353 and Q384). In all cases, the Km of the enzyme variant was higher than that of GOOX, supporting the role of corresponding residues in substrate binding. Most notably, W351A increased Km values by up to two orders of magnitude while also increasing kcat up to 3-fold on cello- and xylo-oligosaccharides and showing no substrate inhibition.ConclusionsThis study provides further evidence that S. strictum GOOX has broader substrate specificity than the enzyme name implies, and that substrate inhibition can be reduced by removing aromatic side chains in the -2 binding subsite. Of the enzyme variants, W351A might be particularly advantageous when oxidizing oligosaccharides present at high substrate concentrations often experienced in industrial processes.

Acid-catalyzed vapor phase esterification with maleic anhydride was used to improve the integrity and thermo-mechanical properties of fiber webs based on poly(vinyl alcohol), PVA. The fibers were produced by electrospinning PVA from aqueous dispersions containing cellulose nanocrystals (CNCs). The effect of esterification and CNC loading on the structure and solvent resistance of the electrospun fibers was investigated. Chemical characterization of the fibers (FTIR, NMR) indicated the formation of ester bonds between hydroxyl groups belonging to neighboring molecules. Thermomechanical properties after chemical modification were analyzed using thermal gravimetric analysis, differential scanning calorimetry, and dynamic mechanical analysis. An 80% improvement in the ultimate strength was achieved for CNC-loaded, crosslinked PVA fiber webs measured at 90% air relative humidity. Besides the ultra-high surface area, the composite PVA fiber webs were water resistant and presented excellent mechanical properties.