Saccharomyces recombinant protein

Yeast expression vectors have been among those most commonly used since the beginning of gene technology. Vectors based on baker s yeast, Saccharomyces cerevisiae, have been especially popular for robust expression of many types of recombinant proteins [90]. For instance, the first commercially available recombinant vaccine, the hepatitis B surface antigen vaccine, was produced from an S. cerevisiae vector [91]. Many other recombinant proteins have also been efficiently expressed in yeast including al-Antitrypsin [92], insulin [93], Epstein-Barr virus envelope protein [94], superoxide dismutase [95] and interferon-a [90]. 

Today, recombinant protein production involves many options. In addition to E. coli, several yeast systems (see Part IV, Chapter 13), insect cells (see Part IV, Chapter 14), different mammalian expression systems (CHO, BHK, NSO, HKBll, PER.C6) (see Part II, Chapter 3 and Part IV, Chapters 1 and 3) other alternative expression systems are currently under development for the production of biopharmaceuticals. These include transgenic animals or plants, and will be discussed in Part IV, Sub-Part 2 of this book. This chapter will focus on E. coli, a still-modern secretory Saccharomyces ccrevisiac system, and the recently developed mammalian HKBll expression system. An E. coli host/vector system is described that was originally developed for the efficient production of an interleukin-4 variant Later, it transpired that this system is ideally suited to the expression of other proteins and Fab fragments. The secretory... 

Yeast display uses the a-agglutinin receptor to display recombinant proteins on the surface of Saccharomyces cerevisiae. Yeast display of scFvs or Fabs allows the detection and selection by fluorescence-assisted flow cytometry or by magnetic sorting. In addition, flow cytometry can be used for kinetic characterization of antibody affinity (K ) as well as K ff and rates. ... 

Topical anti-ulcer. Recombinant protein produced in the yeast, Saccharomyces cerevisiae. Homodimer. 

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. 

Chi WK, Ku CH, Chang CC (1994) Two-step cell disruption for the extraction of membrane-associated recombinant protein from Saccharomyces cerevisiae. Ann NY Acad Sci 721(1) 365-373... 

Yeast was the first microbial host used by mankind for biotransformation of raw materials, and it marked the early developments of industrial biotechnology. Initially, Saccharomyces cerevisiae and closely related species were used because of their high fermentative capacity and based on the vast experience from alcoholic beverage production. While a high fermentation rate is favorable for the production of bioethanol and other primary metabolites, it implicates disadvantages for growth-coupled production. Consequently, a number of other yeasts have been developed for the production of biofuels, biochemicals, lipids, or recombinant proteins. 

R)- and ( -selective HNLs. A number of recombinant HNLs have also been expressed in E. coli, Saccharomyces cerevisiae, and Pichia pastoris. Recently, protein engineering has been successfully applied to the development of a tailor-made HNL for large-scale production of specific cyanohydrins [69,70]. 

Human Set9 is a 50 kDa H3 methyltransferase that methylates Lys-4 of H3. The enzyme methylated free H3 but not H3 in chromatin substrates. There is evidence that Set9 may stimulate activated transcription [198]. Set9 has the SET domain but lacks the cysteine-rich (pre-SET and post-SET) domains. Disruption of Saccharomyces cerevisiae and Saccharomyces pombe Setl obliterates H3 methyl Lys-4 [199]. Thus this SET domain containing protein appears to be a H3 Lys-4 methyltransferase, catalyzing both di- and tri-methylation of H3 Lys-4 [155]. However, studies with recombinant Setl failed to show histone methyltransferase activity. It has been suggested that other associated proteins may be required for the Setl to be catalytically active [139,200]. Indeed, Setl is associated with several... 

There exist a variety of vectors for cloning into eukaryotic systems, ranging from yeast (Saccharomyces as well as Pichia) through insect cells (Baculovims) and plants (Ti plasmid from Agrobacterium tumefaciens) to mammalian cells (transfected by viral or mammalian vectors). As expression in eukaryotic hosts is less efficient than bacterial expression in terms of yield and time and more complicated in terms of vector structure and culture conditions, such eukaryotic expression systems are only used for genes whose proteins require posttranslational modification which is not possible in bacteria. Yeast is the preferred option as a relatively easily culturable single-cell system but posttranslational modification capabilities is limited. The additional complexity can be circumvented in part by exploiting the ability of eukaryotic vectors to act as shuttle vectors, which can be shuttled between two evolutionarily different hosts. Thus, eukaryotic vectors can be replicated and analyzed in bacteria and transfected into eukaryotic cells for expression of the recombinant product. 

Brand MD, Buckingham JA, Esteves TC, Green K, Lambert AJ, Miwa S, Murphy MP, Pakay JL, Talbot DA, Echtay KS (2004) Mitochondrial superoxide and aging uncoupling-protein activity and superoxide production. Biochem Soc Symp 71 203-213 Brennan RJ, Schiestl RH (1998) Chloroform and carbon tetrachloride induce intrachromosomal recombination and oxidative free radicals in Saccharomyces cerevisiae. Mutat Res 397 271-278... 

Blanquet, S., Antonelli, R., Laforet, L., Denis, S., Marol-Bonnin, S., and Alric, M. (2004), Living recombinant Saccharomyces cerevisiae secreting proteins or peptides as a new drug delivery system in the gut, J. Biotechnol., 110, 37-49. 

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