Upon oxidation of fibers with laccase, Mw and Mn of the dioxane-soluble lignin increased 40 and 20%, respectively, giving only a minor change in polydispersity. Hot pressing of laccase treated fibers into fiberboards causes a substantial lignin polymerization, compared to pressing of untreated fibers. From the shoulder in the chromatogram in Fig. 3B, the Mw was increased about 70%, whereas Mn increased about 20%. The polydispersity increased from 11.7 to 16.2. For boards only, a small residue of the dioxane-extractable lignin from the enzyme- and control-treatments turned out to be insoluble in THF. The decreased solubility could be caused by a heat-induced polymerization of lignin, perhaps by cross-linking to carbohydrates.
The increased molecular mass of isolated surface lignin from the laccase treated boards strongly suggests that polymerization of lignin is part of the increased bonding, i.e. covalent bonding of the lignin is taking place. The question is whether lignin tightly associated to fibers is involved in the polymerization, or if the strength is gained solely from polymerization of loosely associated lignin. Since technical lignin is known to have significant improved adhesive properties when polymerized through a laccase/peroxidase catalyzed oxidation [1], obviously the polymerization of lignin loosely associated to fibers would be expected to have a similar effect. This may also be supported from the fact that equal levels of radicals are seen for 6 and 24 U, but the strength of boards treated with 24 U is higher. This could indicate that, e.g. precipitation of lignin on the fiber surfaces and thereby loosely associated lignin is partly responsible for the bonding effect.
Given that the dimensional properties of the boards using laccase as bonding catalyst can be improved to a level comparable to synthetic adhesives, the described process may be scaled up for full industrial application. The nature of laccase catalyzed bonding requires higher pressing temperatures and longer pressing times, two reasons why the concept may not be economically feasible as it is. However, the concept shows promise and possibilities in the use of oxidative enzymes for industrial bonding and modification of lignin.
4. Conclusion
Fiberboards bonded by laccase catalyzed oxidation can be made in a pilot-scale process simulating continuous full-scale industrial production. The strength properties of the enzyme bonded boards are comparable to boards bonded by a conventional UF-resin and wax, whereas the dimensional stability properties of the enzyme bonded boards are not at the same level. The latter cannot be improved using a wax as this inhibits the bonding effect of the enzyme. This is most likely caused by reduced fiber–fiber interactions caused by a wax coating of the fibers. In enzyme bonded boards a cross-linking of the lignin can be observed. It cannot be concluded if this cross-linking is forming covalent bonds between fibers or if the polymerized loosely associated lignin works as an adhesive.
Acknowledgements
This work was financed by the Danish Ministry of Environment, the grant committee for product development in the forestry and timber industries, and Novozymes A/S.