title: Characterization of five terminator regions that increase the protein yield of a transgene in Saccharomyces cerevisiae.
abstract: Strong terminator regions could be used to improve metabolically engineered yeasts by increasing the target enzyme protein yields above those achieved with traditional terminator regions. We recently identified five strong terminator regions (RPL41Bt, RPL15At, DIT1t, RPL3t, and IDP1t) in a comprehensive analysis of Saccharomyces cerevisiae. The effect of the terminator regions was analyzed by measuring the protein production of a linked transgene, and was shown to be twice that of a traditional terminator region (PGK1t). Here, we investigated whether the activity of the terminator regions is affected by exchange of a strong promoter or reporter in the linked transgene, carbon source for cell growth, stress factors, host yeast strain, or stage of the growth phase. Our results indicate that the activities of all five terminator regions were twice that of PGK1t in all conditions tested. In addition, we demonstrated that the strong activity of these terminator regions could be used to improve secretory production of endoglucanase II derived from Tricoderma ressei, and that the DIT1t strain was the best of the five strains for this purpose. We therefore propose that DIT1t, and the four other terminator regions, could be applied to the development of improved metabolically engineered yeasts.
authors.affiliation: Matsuyama Research Group, Toyota Central Research and Development Laboratories, Incorporated, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Ito
authors.forename: Mamoru
authors.initials: M
authors.lastname: Yamanishi
authors.forename: Akinori
authors.initials: A
authors.lastname: Ikeuchi
authors.forename: Chie
authors.initials: C
authors.lastname: Imamura
authors.forename: Kenro
authors.initials: K
authors.lastname: Tokuhiro
authors.forename: Takao
authors.initials: T
authors.lastname: Kitagawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsuyama
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D019149:Bioreactors; D002244:Carbon / Q000737:chemistry / Q000378:metabolism; D002480:Cellulase / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016681:Genome, Fungal; D060847:Metabolic Engineering; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D013728:Terminator Regions, Genetic; D019076:Transgenes
chemical_list: D020413:3' Untranslated Regions; D002244:Carbon; C056373:endoglucanase 2; D002480:Cellulase
keywords: 3′-UTR activity; Cellulase; FI; Fluorescent protein; Metabolic engineering; RP; SS; Terminator region; codon-optimized green fluorescent protein; fluorescence intensity; opGFP; ribosomal protein; side-scatter intensity