Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae |
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| Institution: | 1. Novo Nordisk Foundation Center for Biosustainability, Department of Chemical and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE-41296 Göteborg, Sweden;2. Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Fremtidsvej 3, DK-2970 Hørsholm, Denmark;3. CNRS, Institut de Biochimie et Génétique Cellulaires, UMR 5095, F-33000 Bordeaux, France;4. Université de Bordeaux, Institut de Biochimie et Génétique Cellulaires, UMR 5095, F-33000 Bordeaux, France;1. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States;2. Department of Chemical and Biomolecular Engineering, Tulane University, New Orleans, LA 70118, United States;1. Department of Agricultural and Biological Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA;2. Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL, USA;3. Bioenergy Research Unit, National Center for Agricultural Utilization Research, USDA-ARS, Peoria, IL 61604, USA;1. Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA, 61801;2. Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, IL, 61801, USA;3. Beckman Institute for Advanced Science & Technology, University of Illinois at Urbana-Champaign, IL, 61801, USA;1. Micronutrients Laboratory, Institute of Nutrition and Food Technology (INTA), University of Chile, Avda. El Líbano 5524, Santiago, Chile;2. Faculty of Veterinary and Animal Sciences, University of Chile, Santa Rosa 11.735, La Pintana, Santiago, Chile;3. United States Department of Agriculture, Western Human Nutrition Research Center, University of California, 430 W. Health Sciences Drive, Davis, CA 95616, USA;4. Department of Food Science and Technology, University of California, 1 Shields Avenue, University of California, Davis, CA 95616, USA;1. Department of Bio and Fermentation Convergence Technology, and Interdisciplinary Program for Bio-health Convergence, Kookmin University, Seoul 02707, Republic of Korea;2. Center for Industrialization of Agricultural and Livestock Microorganism (CIALM), Jeongeup 56212, Republic of Korea;3. Department of Food Science and Technology, Chung-Ang University, Anseong, Gyeonggi 17546, Republic of Korea;4. Department of Integrative Food, Bioscience and Biotechnology, Chonnam National University, Gwangju 61186, Republic of Korea |
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| Abstract: | Due to limitations associated with whole blood for transfusions (antigen compatibility, transmission of infections, supply and storage), the use of cell-free hemoglobin as an oxygen carrier substitute has been in the center of research interest for decades. Human hemoglobin has previously been synthesized in yeast, however the challenge is to balance the expression of the two different globin subunits, as well as the supply of the prosthetic heme required for obtaining the active hemoglobin (α2β2). In this work we evaluated the expression of different combinations of α and β peptides and combined this with metabolic engineering of the heme biosynthetic pathway. Through evaluation of several different strategies we showed that engineering the biosynthesis pathway can substantially increase the heme level in yeast cells, and this resulted in a significant enhancement of human hemoglobin production. Besides demonstration of improved hemoglobin production our work demonstrates a novel strategy for improving the production of complex proteins, especially multimers with a prosthetic group. |
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| Keywords: | Human hemoglobin Heme biosynthesis Prosthetic group Protein production |
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