Improved production of human hemoglobin in yeast by engineering hemoglobin degradation

Olena P. Ishchuk*, August Thinggaard Frost, Facundo Muñiz-Paredes, Saki Matsumoto, Nathalie Laforge, Nélida Leiva Eriksson, José L. Martínez, Dina Petranovic*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

With the increasing demand for blood transfusions, the production of human hemoglobin (Hb) from sustainable sources is increasingly studied. Microbial production is an attractive option, as it may provide a cheap, safe, and reliable source of this protein. To increase the production of human hemoglobin by the yeast Saccharomyces cerevisiae, the degradation of Hb was reduced through several approaches. The deletion of the genes HMX1 (encoding heme oxygenase), VPS10 (encoding receptor for vacuolar proteases), PEP4 (encoding vacuolar proteinase A), ROX1 (encoding heme-dependent repressor of hypoxic genes) and the overexpression of the HEM3 (encoding porphobilinogen deaminase) and the AHSP (encoding human alpha-hemoglobin-stabilizing protein) genes — these changes reduced heme and Hb degradation and improved heme and Hb production. The reduced hemoglobin degradation was validated by a bilirubin biosensor. During glucose fermentation, the engineered strains produced 18% of intracellular Hb relative to the total yeast protein, which is the highest production of human hemoglobin reported in yeast. This increased hemoglobin production was accompanied with an increased oxygen consumption rate and an increased glycerol yield, which (we speculate) is the yeast's response to rebalance its NADH levels under conditions of oxygen limitation and increased protein-production.
Original languageEnglish
JournalMetabolic Engineering
Volume66
Pages (from-to)259-267
Number of pages9
ISSN1096-7176
DOIs
Publication statusPublished - 2021

Keywords

  • Human hemoglobin
  • Heme
  • Saccharomyces cerevisiae
  • Reduced degradation
  • Bilirubin biosensor

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