Abstract
Lignocellulose is composed of three principal plant cell wall polymers (cellulose, hemicellulose and lignin) in a closely knit compound matrix, which promotes stability and renders the component polysaccharides insoluble. Lignocellulose is therefore refractory to plant cell wall degrading enzymes that can be used in biorefinary applications, such as biobleaching and the production of fermentable sugars from plant cell-wall biomass. As a consequence of lignocellulose complexity, individual enzymes modify the constituent polymers inefficiently, and biorefinary applications will require mixtures of enzymes with different specificities. The current proposal aims to identify, characterize and engineer the catalytic properties of enzymes with the goal of developing a portfolio of biocatalysts that may be combined to form enzymatic cocktails for the biorefinary of lignocellulose. The overall strategy will focus on the characterization of enzymes identified in genomic sequencing projects of the fungi Aspergillus niveus, Aspergillus japonicus and Chaetomium thermophilum and the bacteria Bacillus subtilis and Clostridium thermocellum, using a battery of microbiological and biochemical techniques coupled with molecular structural biology approaches. This initial targeting and biochemical characterization effort will be undertaken in parallel with more sophisticated analyses which will identify in finer detail the exact enzymatic modifications of the plant cell wall polymers. These activities will provide information for a vigorous protein engineering program using rational protein design and directed evolution strategies which will aim to improve the catalytic properties of selected enzymes for biorefinary applications in the cellulose pulp and paper industries and in the development of biofuel technologies. (AU)
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