Improved production of human hemoglobin in yeast by engineering hemoglobin degradation
Ishchuk, Olena P. LU ; Frost, August T. ; Muñiz-Paredes, Facundo ; Matsumoto, Saki ; Laforge, Nathalie ; Eriksson, Nélida Leiva LU
; Martínez, José L.
and Petranovic, Dina
(2021)
In Metabolic Engineering
66.
p.259-267
- 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... (More)
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.
(Less)
- author
- Ishchuk, Olena P.
LU
; Frost, August T.
; Muñiz-Paredes, Facundo
; Matsumoto, Saki
; Laforge, Nathalie
; Eriksson, Nélida Leiva
LU
; Martínez, José L.
and Petranovic, Dina
- organization
- publishing date
- 2021-07
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Bilirubin biosensor, Heme, Human hemoglobin, Reduced degradation, Saccharomyces cerevisiae
- in
- Metabolic Engineering
- volume
- 66
- pages
- 9 pages
- publisher
- Elsevier
- external identifiers
-
- scopus:85105798047
- pmid:33984513
- ISSN
- 1096-7176
- DOI
- 10.1016/j.ymben.2021.05.002
- language
- English
- LU publication?
- yes
- id
- c032e300-4ff7-4e7e-9dc1-e17813bf8e73
- date added to LUP
- 2021-05-27 16:21:28
- date last changed
- 2024-05-05 06:34:18
@article{c032e300-4ff7-4e7e-9dc1-e17813bf8e73,
abstract = {{<p>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.</p>}},
author = {{Ishchuk, Olena P. and Frost, August T. and Muñiz-Paredes, Facundo and Matsumoto, Saki and Laforge, Nathalie and Eriksson, Nélida Leiva and Martínez, José L. and Petranovic, Dina}},
issn = {{1096-7176}},
keywords = {{Bilirubin biosensor; Heme; Human hemoglobin; Reduced degradation; Saccharomyces cerevisiae}},
language = {{eng}},
pages = {{259--267}},
publisher = {{Elsevier}},
series = {{Metabolic Engineering}},
title = {{Improved production of human hemoglobin in yeast by engineering hemoglobin degradation}},
url = {{http://dx.doi.org/10.1016/j.ymben.2021.05.002}},
doi = {{10.1016/j.ymben.2021.05.002}},
volume = {{66}},
year = {{2021}},
}