title: Expression and high-level secretion of Trichoderma reesei endoglucanase I in Yarrowia lipolytica.
abstract: The endoglucanase I (EGI) from fungus Trichoderma reesei was cloned, expressed, and secreted from Yarrowia lipolytica using the XPR2 promoter. The signal sequence of EGI transferred from T. reesei was efficiently processed in the Y. lipolytica secretory pathway and directed the secretion of active EGI into the culture medium. However, the recombinant EGI produced from YLCSIn strain was hyperglycosylated and significantly larger than the native enzyme produced by the parent strain. The expression of EGI using XPR2 preproregion has caused secretion of modified proteins that still retained cellulase activity. This resulted from imprecise processing of the N-terminus of recombinant protein. While the batch culture produced 5 mg EGI/L from YLCSIn strain, the EGI yield was increased approx 20-fold when the fed-batch fermentation process strategy in combination with the high-cell density cultivation technique was employed. These results showed that the Y. lipolytica is a useful host organism for production of a large amount of large size heterologous proteins, especially when used in combination with high-cell density and fed-batch culture techniques.
authors.affiliation: Department of Food Science and Technology, University of California, Davis 95616, USA.
authors.forename: C S
authors.initials: CS
authors.lastname: Park
authors.forename: C C
authors.initials: CC
authors.lastname: Chang
authors.forename: D D
authors.initials: DD
authors.lastname: Ryu
mesh_terms: D002480:Cellulase / Q000235:genetics* / Q000378:metabolism*; D043366:Cellulose 1,4-beta-Cellobiosidase; D005285:Fermentation; D015870:Gene Expression; D005822:Genetic Vectors; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D014162:Transfection; D014242:Trichoderma / Q000201:enzymology*; D015003:Yeasts / Q000201:enzymology*
chemical_list: D011994:Recombinant Proteins; D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase