Investigation into the Production of a Cellulytic Saccharomyces – Identification of a High Cellulytic Trichoderma spp. for Gene Selection

The isolation and identification of a hyper-cellulytic fungus for cellulase gene selection was carried out as the initial stages in the production of a recombinant micro-organism. This would be capable of Consolidated BioProcessing (CBP) of cellulosic material to ethanol in single stage fermentations. With the selection of an organism with high cellulytic rate, the construction of a cellulytic ethanogen through the expression of fungal cellulosic genes in an Saccharomyces spp. would be seen as the second stage. This study focus looked at the isolation and identification of a hyper cellulytic Trichoderma spp.. For the selection of fungal genes, a Trichoderma species with a high cellulytic rate was isolated from pine forest litter from North Wales, UK. Using metabolite selection techniques, the cellulytic rate was identified with 18s rDNA analysis used for strain identification. HPLC analysis confirmed celluyltic rate in comparison to the hyper-cellulytic strain T. reesei RutC30. 18s rDNA analysis and sequencing, returning a reading frame indicating 98% homology with Trichoderma with no complete matches, indicating the possibility of unique strain isolation. Further analysis would be required for confirmation. Celluyltic rate was seen to be greater than RutC30 with a 30% increase.