Thermoascus aurantiacus CBHl/Cel7A Production in Trichoderma reesei on Alternative Carbon Sources

摘要

To develop functional enzymes in cellulose hydrolysis at or above 70°C the cellobiohydrolase (CBHI/Cel7A) of Thermoascus aurantiacus was cloned and expressed in Trichoderma reesei Rut-C30 under the strong cbhl promoter. Cellulase production of the parental strain and the novel strain (RF6026) was examined in submerged fermentation experiments using various carbon sources, which were lactose, Solka Floe 200 cellulose powder, and steam pre-treated corn stover. An industrially feasible production medium was used containing only distiller's spent grain, KH_2PO_4, and (NH_4)_2SO_4. Enzyme production was followed by measurements of protein concentration, total cellulase enzyme activity (filter paper activity), p-glucosidase activity, CBHI activity, and endogenase I (EGI) activity. The Thermoascus CBHI/Cel7A activity was taken as an indication of the heterologous gene expression under the cbhl promoter.