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Synthesis of Wild Type and Mutant Human Hemoglobins in Saccharomyces cerevisiae

Bio/Technology volume 9, pages 57–61 (1991)Cite this article

Abstract

We have expressed human αand β-globin cDNA clones from separate, synthetic galactose-regulated hybrid promoters contained on a single plasmid in Saccharomy-ces cerevisiae. Co-expression of the α and β-globin chains in S. cerevisiae results in the assembly of these proteins into soluble tetrameric hemoglobin that accumulates to 3–5 percent of the total cell protein. Endogenously produced heme is incorporated into the tetramer and the protein produced is functionally and structurally indistinguishable from human Ao hemoglobin. This expression system has been used to produce both wild type hemoglobin and a low O2-affinity hemoglobin mutant that has oxygen binding and dissociation characteristics similar to human whole blood. The yeast expression system we describe may be suitable for the production of a recombinant hemoglobin based blood substitute as well as for detailed structure-activity studies of human hemoglobin.

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Author notes

  1. Gary Stetler: Corresponding author.

Authors and Affiliations

  1. Somatogen, Inc., 350 Interlocken Parkway, Broomfield, CO, 80020

    Michael Wagenbach, Katherine O'Rourke, Laura Vitez, Anna Wieczorek, Stephen Hoffman, Steven Durfee, John Tedesco & Gary Stetler

Authors
  1. Michael Wagenbach
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  2. Katherine O'Rourke
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  5. Stephen Hoffman
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  7. John Tedesco
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  8. Gary Stetler
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Wagenbach, M., O'Rourke, K., Vitez, L. et al. Synthesis of Wild Type and Mutant Human Hemoglobins in Saccharomyces cerevisiae. Nat Biotechnol 9, 57–61 (1991). https://doi.org/10.1038/nbt0191-57

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