Improving the Production of Cofactor-Containing Proteins: Production of Human Hemoglobin in Yeast

Ishchuk, Olena P.; Martínez, José Luis; Petranović, Dina

doi:10.1007/978-1-4939-9024-5_11

Cited by 3 publications

(2 citation statements)

References 76 publications

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“…Besides the experimentally validated the predicted engineering targets for the production of αamylase (Fig. 6), we also noticed the consistence of the predicted targets for other recombinant proteins with literature reports, such as Hem2, Hem3 and Hem12 for hemoglobin production 28,51 . We also confirmed that even though Hem4 is also in the heme synthesis pathway, this is not a rate-limiting step in the heme synthesis 51 .…”

Section: Discussionsupporting

confidence: 71%

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Genome scale modeling of the protein secretory pathway reveals novel targets for improved recombinant protein production in yeast

et al. 2021

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Eukaryal cells are used for the production of many recombinant pharmaceutical proteins, including several of the current top-selling products. The protein secretory pathway in eukaryal cells is complex and involves many different processes such as post-translational modifications, translocation, and folding. Furthermore, recombinant protein production competes with native secretory proteins for the limited energy and proteome resources allocated to the protein secretory pathway. Due to the complexity of this pathway, improvement through metabolic engineering has traditionally been relatively ad-hoc; and considering the industrial importance of this pathway, there is a need for more systematic approaches for novel design principles. Here, we present the first proteome-constrained genome-scale protein secretory model of a eukaryal cell, namely for the yeast Saccharomyces cerevisiae (pcSecYeast). The model contains all key processes of this pathway, i.e., protein translation, modification, and degradation coupled with metabolism. The model can capture delicate phenotypic changes such as the switch in the use of specific glucose transporters in response to changing extracellular glucose concentration. Furthermore, the model can also simulate the effects of protein misfolding on cellular growth, suggesting that retro-translocation of misfolded proteins contributes to protein retention in the Endoplasmic reticulum (ER). We used pcSecYeast to simulate various recombinant proteins production and identified overexpression targets for different recombinant proteins overproduction. We experimentally validated many of the predicted targets for α-amylase production in this study, and the results show that the secretory pathways have more limited capacity than metabolism in terms of protein secretion.

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Section: Discussionsupporting

confidence: 71%

“…Note that hemoglobin folds with heme as a prosthetic group, which requires balancing of heme biosynthesis and its recombinant protein production (Fig. 4a) 28 . We generated eight specific models to simulate the maximum recombinant protein secretion under various growth rates.…”

Section: Protein Features Impact Recombinant Protein Productionmentioning

confidence: 99%

Genome scale modeling of the protein secretory pathway reveals novel targets for improved recombinant protein production in yeast

et al. 2021

Preprint

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Classifications of Blood Substitutes

Liu

Kaye

Verbeek

et al. 2022

Blood Substitutes and Oxygen Biotherapeutics

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Translational Research of Hemoglobin Vesicles as a Transfusion Alternative

Sakai

Kobayashi

Kure

et al. 2022

CMC

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: Clinical situations arise in which blood for transfusion becomes scarce or unavailable. Considerable demand for a transfusion alternative persists because of various difficulties posed by blood donation and transfusion systems. Hemoglobin-vesicles (HbV) are artificial oxygen carriers being developed for use as a transfusion alternative. Just as biomembranes of red blood cells (RBCs) do, phospholipid vesicles (liposomes) for Hb encapsulation can protect the human body from toxic effects of molecular Hb. The main HbV component, Hb, is obtained from discarded human donated blood. Therefore, HbV can be categorized as a biologic agent targeting oxygen for peripheral tissues. The purification procedure strictly eliminates the possibility of viral contamination. It also removes all concomitant unstable enzymes present in RBC for utmost safety from infection. The deoxygenated HbVs, which are storable for over years at ambient temperature, can function as an alternative to blood transfusion for resuscitation from hemorrhagic shock and O2 therapeutics. Moreover, a recent study clarified beneficial effects for anti-oxidation and anti-inflammation by carbon monoxide (CO)-bound HbVs. Autoxidation of HbV (HbO2 → metHb + O2-.) is unavoidable after intravenous administration. Co-injection of methylene blue can extract the intraerythrocytic glycolytic electron energy effectively and reduce metHb. Other phenothiazine dyes can also function as electron mediators to improve the functional life span of HbV. This review paper summarizes recent progress of the research and development of HbV, aimed at clinical applications.

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Improving the Production of Cofactor-Containing Proteins: Production of Human Hemoglobin in Yeast

Cited by 3 publications

References 76 publications

Genome scale modeling of the protein secretory pathway reveals novel targets for improved recombinant protein production in yeast

Genome scale modeling of the protein secretory pathway reveals novel targets for improved recombinant protein production in yeast

Classifications of Blood Substitutes

Translational Research of Hemoglobin Vesicles as a Transfusion Alternative

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