Biomass Depolymerization projects

Development of Neurospora crassa as an Expression Platform and Model Organism to Explore Functional Analysis of Celluloytic Enzymes - Completed

The project began developing the tools available for N. crassa, a well-established model for laboratory studies with a large toolbox of molecular, genomics, and cytological techniques, to understand how secretion of plant cell wall degrading enzymes is regulated. In mapping out an engineering strategy for any filamentous fungus to achieve maximal amounts of secreted enzymes for industrial use, the group set the foundation for further EBI studies into the enzymes' role in lignocellulosic deconstruction of plant material for biofuel production.

project Highlights

2011 Highlights

Glass is looking at ways that Neurospora crassa regulates lignocellulose deconstruction and secretion of lignocellulolytic enzymes with an eye towards protein and metabolic engineering. To identify transcriptional regulators of biomass-degrading enzymes, her group screened over 200 mutants and is characterizing about 20 strains with altered enzyme activity or growth on cellulose. Her group also designed tools to probe the secretory pathway that may yield tools to increase cellulolytic enzyme output.

2010 Highlights

Glass’s group successfully expressed a functional GFP-tagged endoglucanase II protein and showed that this protein binds to intact plant cell walls.  The strain is also being used to assess trafficking of cellulolytic and hemicellulolytic enzymes.

2009 Highlights

Glass and her team purified and characterized catalytically active proteins representing endoglucanase II, endoglucanase IV, cellobiose dehydrogenase I, and cellobiose hydrolase. They also are investigating two secreted proteins of unknown function that affect cellulase activity. Furthermore, Glass’s team is developing strains to facilitate over-expression of cellulases, hemicellulases and proteins of unknown function. Their goal is to construct vectors and strains that allow high production of secreted proteins that will be evaluated for biochemical, structural and enzymatic activities involved in plant cell wall deconstruction.

 


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