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

• Published in: Applied biochemistry and biotechnology 2007-04, Vol.137-140 (1-12), p.195-204
• Main Authors: BENKÖ, Zsuzsa, DRAHOS, Eszter, SZENGYEL, Zsolt, PURANEN, Terhi, VEHMAANPERÄ, Jar, RECZEY, Kati
• Format: Article
• Language: English
• Subjects: Ascomycota - genetics; Ascomycota - metabolism; Biological and medical sciences; Bioreactors; Biotechnology; Carbon; Carbon Compounds, Inorganic - metabolism; Carbon sources; Cellulose; Cellulose 1,4-beta-Cellobiosidase - genetics; Cellulose 1,4-beta-Cellobiosidase - metabolism; Cloning; Enzymatic activity; Enzymes; Fermentation; Fundamental and applied biological sciences. Psychology; Fungi; Genetic Enhancement - methods; Hypocrea jecorina; Lactose - metabolism; Methods. Procedures. Technologies; Protein Engineering - methods; Stover; Studies; Thermoascus aurantiacus; Trichoderma - genetics; Trichoderma - metabolism; Various methods and equipments; Zea mays - microbiology
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-007-9051-5

To develop functional enzymes in cellulose hydrolysis at or above 70 degrees C the cellobiohydrolase (CBHI/Cel7A) of Thermoascus aurantiacus was cloned and expressed in Trichoderma reesei Rut-C30 under the strong cbh1 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 Floc 200 cellulose powder, and steam pretreated corn stover. An industrially feasible production medium was used containing only distiller's spent grain, KH(2)PO(4), and (NH(4))(2)SO(4). Enzyme production was followed by measurements of protein concentration, total cellulase enzyme activity (filter paper activity), beta-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 cbh1 promoter.