Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae

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 synth...

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Main Authors: Liu, Lifang, Martínez, J., Liu, Z., Petranovic, D., Nielsen, J.
Format: Journal Article
Published: Academic Press Inc. 2014
Online Access:http://hdl.handle.net/20.500.11937/22603
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author Liu, Lifang
Martínez, J.
Liu, Z.
Petranovic, D.
Nielsen, J.
author_facet Liu, Lifang
Martínez, J.
Liu, Z.
Petranovic, D.
Nielsen, J.
author_sort Liu, Lifang
building Curtin Institutional Repository
collection Online Access
description 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 (a2ß2). In this work we evaluated the expression of different combinations of a and ß 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. © 2013 .
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publishDate 2014
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spelling curtin-20.500.11937-226032018-03-29T09:06:34Z Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae Liu, Lifang Martínez, J. Liu, Z. Petranovic, D. Nielsen, J. 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 (a2ß2). In this work we evaluated the expression of different combinations of a and ß 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. © 2013 . 2014 Journal Article http://hdl.handle.net/20.500.11937/22603 10.1016/j.ymben.2013.10.010 Academic Press Inc. restricted
spellingShingle Liu, Lifang
Martínez, J.
Liu, Z.
Petranovic, D.
Nielsen, J.
Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae
title Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae
title_full Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae
title_fullStr Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae
title_full_unstemmed Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae
title_short Balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in Saccharomyces cerevisiae
title_sort balanced globin protein expression and heme biosynthesis improve production of human hemoglobin in saccharomyces cerevisiae
url http://hdl.handle.net/20.500.11937/22603