Saccharomyces cerevisiae glycosylation

The glycosylated derivatives 51A-D were tested against the yeast Saccharomyces cerevisiae and displayed different photoactivities.24 In this case, the most photoactive compounds were the mono-glycosylated 51A and... 

Caro, L., Tettelin, H., Vossen, J., Ram, A., van den Ende, H., and Klis, F. (1997). In silico identification of glycosyl-phosphatidylinositol-anchored plasma-membrane and cell wall proteins of Saccharomyces cerevisiae. Yeast 13, 1477—1489. 

Saccharomyces cerevisiae processing High yields (g/l in Pichia pastoris) Inexpensive Formation of disulphides Glycosylation mammalian systems... 

Mannose is primarily found in mammalian N-linked glycoproteins as part of core structures containing the tri-Man core [a-D-Man-(1 3)-a-D-Man-(1 6)-Man]. However, in the glycoproteins of yeast and molds, mannose, primarily a-D-Man-(1 2)-a-D-Man and a-D-Man-(1 6)-a-D-Man, has been found a-linked to Ser or Thr. 244 Expression of glucoamylase G1 in Aspergillus niger 245 and human IGF-I (somatomedin C) in Saccharomyces cerevisiae 246 resulted in proteins glycosylated with mannose at Ser and Thr. 

According to a review by Walsh [10], of 165 biopharmaceuhcal products approved in the United States and Europe by 2006, only two are nucleic acid-based drugs, whereas nine of the 31 therapeutic proteins approved since 2003 are produced in E, coli, and 17 are produced by mammalian cell lines. In 2004 market distribution and manufacture of therapeutic proteins, non-glycosylated (non-antibody) proteins constitutes 40% of the total market, with 12% armual growth rate, and are produced in E. coli or the yeast Saccharomyces cerevisiae glycoproteins (primarily mAbs) constitute 60% of the total market, with 26% armual growth rate, and are produced by mammalian cell culture (mostly with cells from Chinese Hamster Ovary, or CHO). 

Ballou, C. E. (1990). Isolation, characterization, and properties of Saccharomyces cerevisiae mann mutants with nonconditional protein glycosylation defects. In Goeddel, DV, editor. Methods in Enzymology, Vol. 185, Academic Press, San Diego, Calif., pp.440-470. 

Zoecklein, B.W., Marcy, J.E., Williams, J.M. Jasinsky, Y. (1997b). Effect of native strain of Saccharomyces cerevisiae on glycosyl-glucose, potential volatile terpenes, and selected aglycones of White Riesling (Vitis vinifera L.) wines. J. Eood Comp. Anal., 10, 55-65. 

Gellerfors, P, Axelsson, K, Helander, A, Johansson, S, Kenne, L, Lindqvist, S, Pavlu, B, Skottner, A, Eryklund, L, Isolation and characterization of a glycosylated form of human insuUn-like growth factor I produced in Saccharomyces cerevisiae, J. Biol. Chem., 264, 11444-11449, 1989. 

Studies on the specificities of purified a-D-glycosyl hydrolases of about eight kinds of yeast are summarized in Table XI. The enzymes of Saccharomyces cerevisiae, namely, a-D-glucosidase and oligo-(1 — 6)-D-glucosidase (EC 3.2.1.10), have been examined the most thoroughly. 

Vickers, M.E., Mani, R.S., Sundaram, M., Hogue, D.L., Young, J.D., Baldwin. S.A., and Cass, C.E. (1999) Eunctional production and reconstitution of the human equilibrative nucleoside transporter (hENTl) in Saccharomyces cerevisiae. Interaction of inhibitors of nucleoside transport with recombinant hENTl and a glycosylation-defective derivative (hENTl/N48Q). The Biochemical Journal, ii9 (Pt 1). 21—32,... 

Barnes, C., Hansen, W. J., Holcomb, C. L., and Rine, J. (1984). Asparagine-linked glycosylation in Saccharomyces cerevisiae Genetic analysis of an early step. Mol. Cell Biol. 4, 2381-2388. 

Bernstein, M., Kepes, F., and Schekman, R. (1989). Sec59 encodes a membrane protein required for core glycosylation in Saccharomyces cerevisiae. Mol. Cell Biol. 9,1191—1199. 

Gentzsch, M., and Tanner, W. (1996). The PMT gene family Protein O-glycosylation in Saccharomyces cerevisiae is vital. EMBO J. 15, 5752—5759. 

Expression of recombinant aprotinins has been achieved in E. coli K12 as a fusion with parts of the MS-2 polymerase gene [16]. In this case the fusion protein is deposited as inclusion bodies. Functionally active aprotinin can be obtained after solubilization and purification of the fusion protein, CNBr (cyanobromide) cleavage and renaturation of the aprotinin moiety. Low-level periplasmic expression of native, properly folded aprotinin has been shown in E. coli employing the E. coli alkaline phosphatase signal sequence [17]. With respect to expression level and ease of purification, it transpired that secretory expression in the yeast Saccharomyces cerevisiae is by far the most attractive system for the production of this aprotinin variant [18]. In addition, due to the absence of an N-glycosylation site there are no problems with non-human glycosylation of the protein in yeast. 

Yeast is the third expression system used to produce biopharmaceuticals. As mammalian systems, they possess the ability to cany out post-translational modifications of proteins, although the glycosylation pattern usually varies somewhat from the patterns observed on the native protein or on the protein expressed in mammalian cells. Two recombinant proteins expressed in Saccharomyces cerevisiae are now approved for general medical use hepatitis B surface antigen vaccine and the anticoagulant Hirudin . Alternative promising production systems, in particular transgenic animal and plant systems, are still in development but these systems have to prove that they are technically and economically attractive. 

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