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Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae

Pyruvate and acetyl-coenzyme A, located at the interface between glycolysis and TCA cycle, are important intermediates in yeast metabolism and key precursors for industrially relevant products. Rational engineering of their supply requires knowledge of compensatory reactions that replace predominant...

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Autores principales: van Rossum, Harmen M., Kozak, Barbara U., Niemeijer, Matthijs S., Duine, Hendrik J., Luttik, Marijke A. H., Boer, Viktor M., Kötter, Peter, Daran, Jean-Marc G., van Maris, Antonius J. A., Pronk, Jack T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5815053/
https://www.ncbi.nlm.nih.gov/pubmed/26895788
http://dx.doi.org/10.1093/femsyr/fow017
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author van Rossum, Harmen M.
Kozak, Barbara U.
Niemeijer, Matthijs S.
Duine, Hendrik J.
Luttik, Marijke A. H.
Boer, Viktor M.
Kötter, Peter
Daran, Jean-Marc G.
van Maris, Antonius J. A.
Pronk, Jack T.
author_facet van Rossum, Harmen M.
Kozak, Barbara U.
Niemeijer, Matthijs S.
Duine, Hendrik J.
Luttik, Marijke A. H.
Boer, Viktor M.
Kötter, Peter
Daran, Jean-Marc G.
van Maris, Antonius J. A.
Pronk, Jack T.
author_sort van Rossum, Harmen M.
collection PubMed
description Pyruvate and acetyl-coenzyme A, located at the interface between glycolysis and TCA cycle, are important intermediates in yeast metabolism and key precursors for industrially relevant products. Rational engineering of their supply requires knowledge of compensatory reactions that replace predominant pathways when these are inactivated. This study investigates effects of individual and combined mutations that inactivate the mitochondrial pyruvate-dehydrogenase (PDH) complex, extramitochondrial citrate synthase (Cit2) and mitochondrial CoA-transferase (Ach1) in Saccharomyces cerevisiae. Additionally, strains with a constitutively expressed carnitine shuttle were constructed and analyzed. A predominant role of the PDH complex in linking glycolysis and TCA cycle in glucose-grown batch cultures could be functionally replaced by the combined activity of the cytosolic PDH bypass and Cit2. Strongly impaired growth and a high incidence of respiratory deficiency in pda1Δ ach1Δ strains showed that synthesis of intramitochondrial acetyl-CoA as a metabolic precursor requires activity of either the PDH complex or Ach1. Constitutive overexpression of AGP2, HNM1, YAT2, YAT1, CRC1 and CAT2 enabled the carnitine shuttle to efficiently link glycolysis and TCA cycle in l-carnitine-supplemented, glucose-grown batch cultures. Strains in which all known reactions at the glycolysis-TCA cycle interface were inactivated still grew slowly on glucose, indicating additional flexibility at this key metabolic junction.
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spelling pubmed-58150532018-02-23 Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae van Rossum, Harmen M. Kozak, Barbara U. Niemeijer, Matthijs S. Duine, Hendrik J. Luttik, Marijke A. H. Boer, Viktor M. Kötter, Peter Daran, Jean-Marc G. van Maris, Antonius J. A. Pronk, Jack T. FEMS Yeast Res Research Article Pyruvate and acetyl-coenzyme A, located at the interface between glycolysis and TCA cycle, are important intermediates in yeast metabolism and key precursors for industrially relevant products. Rational engineering of their supply requires knowledge of compensatory reactions that replace predominant pathways when these are inactivated. This study investigates effects of individual and combined mutations that inactivate the mitochondrial pyruvate-dehydrogenase (PDH) complex, extramitochondrial citrate synthase (Cit2) and mitochondrial CoA-transferase (Ach1) in Saccharomyces cerevisiae. Additionally, strains with a constitutively expressed carnitine shuttle were constructed and analyzed. A predominant role of the PDH complex in linking glycolysis and TCA cycle in glucose-grown batch cultures could be functionally replaced by the combined activity of the cytosolic PDH bypass and Cit2. Strongly impaired growth and a high incidence of respiratory deficiency in pda1Δ ach1Δ strains showed that synthesis of intramitochondrial acetyl-CoA as a metabolic precursor requires activity of either the PDH complex or Ach1. Constitutive overexpression of AGP2, HNM1, YAT2, YAT1, CRC1 and CAT2 enabled the carnitine shuttle to efficiently link glycolysis and TCA cycle in l-carnitine-supplemented, glucose-grown batch cultures. Strains in which all known reactions at the glycolysis-TCA cycle interface were inactivated still grew slowly on glucose, indicating additional flexibility at this key metabolic junction. Oxford University Press 2016-02-18 2016-05-01 /pmc/articles/PMC5815053/ /pubmed/26895788 http://dx.doi.org/10.1093/femsyr/fow017 Text en © FEMS 2016. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Research Article
van Rossum, Harmen M.
Kozak, Barbara U.
Niemeijer, Matthijs S.
Duine, Hendrik J.
Luttik, Marijke A. H.
Boer, Viktor M.
Kötter, Peter
Daran, Jean-Marc G.
van Maris, Antonius J. A.
Pronk, Jack T.
Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae
title Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae
title_full Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae
title_fullStr Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae
title_full_unstemmed Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae
title_short Alternative reactions at the interface of glycolysis and citric acid cycle in Saccharomyces cerevisiae
title_sort alternative reactions at the interface of glycolysis and citric acid cycle in saccharomyces cerevisiae
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5815053/
https://www.ncbi.nlm.nih.gov/pubmed/26895788
http://dx.doi.org/10.1093/femsyr/fow017
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