Lars Hildén

Recent developments on cellulases and carbohydrate-binding modules with cellulose affinity

This review concerns basic research on cellulases and cellulose-specific carbohydrate-binding modules (CBMs). As a background, glycosyl hydrolases are also briefly reviewed. The nomenclature of cellulases and CBMs is discussed. The main cellulase-producing organisms and their cellulases are described. Synergy, enantioseparation, cellulases in plants, cellulosomes, cellulases and CBMs as analytical tools and cellulase-like enzymes are also briefly reviewed.

Do the extracellular enzymes cellobiose dehydrogenase and manganese peroxidase form a pathway in lignin biodegradation

The extracellular enzyme manganese peroxidase is believed to degrade lignin by a hydrogen peroxide‐dependent oxidation of Mn(II) to the reactive species Mn(III) that attacks the lignin. However, Mn(III) is not able to directly oxidise the non‐phenolic lignin structures that predominate in native lignin. We show here that pretreatment of a non‐phenolic lignin model compound with another extracellular fungal enzyme, cellobiose dehydrogenase, allows the manganese peroxidase system to oxidise this molecule. The mechanism behind this effect is demethoxylation and/or hydroxylation, i.e. conversion of a non‐phenolic structure to a phenolic one, mediated by hydroxyl radicals generated by cellobiose dehydrogenase. This suggests that cellobiose dehydrogenase and manganese peroxidase may act in an extracellular pathway in fungal lignin biodegradation. Analytical techniques used in this paper are reverse‐phase high‐pressure liquid chromatography, gas chromatography connected to mass spectroscopy and UV‐visible spectroscopy.

Use of a fluorescence labelled, carbohydrate-binding module from Phanerochaete chrysosporium Cel7D for studying wood cell wall ultrastructure

The α-amino group of the carbohydrate-binding module (CBM) from Phanerochaete chrysosporium cellulase Cel7D was covalently labelled with fluorescein isothiocyanate. The fluorescein-labelled CBM was characterised regarding substrate binding, showing specificity only to cellulose and not to mannan and xylan. Conjugation of fluorescein isothiocyanate to CBM did not affect its binding to cellulose. The labelled CBM was successfully used as a probe for detecting cellulose in lignocellulose material such as never dried spruce and birch wood as well as pulp fibres.

Distribution and characterisation of discolouring substances in norway spruce (Picea abies L. Karst.) pulp wood stored under water sprinkling

Abstract Discolouration of wood raw material during wet storage has a notable negative effect on the quality of the final product. In this study, fluorescence microscopy was used to investigate the spatial distribution of discolouring substances in water-sprinkled Norway spruce pulpwood. Water-sprinkled wood was characterised by the presence of discolouring substances on the pit membranes of wood cells. In vitro model studies on the interaction between a model bark substance (tannic acid) and torus constituents support the hypothesis that pectic structures facilitate the allocation of bark substances on pit membranes. Applied pectinase or tannase could not remove the discolouring substances from pit membranes. Manganese peroxidase had a minor but documented effect. The effect of manganese peroxidase, as well as HCl/vanillin labelling, indicated that the discolouring substance may be condensed tannins.