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An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme

Most available knowledge on fungal arginine metabolism is derived from studies on Saccharomyces cerevisiae, in which arginine catabolism is initiated by releasing urea via the arginase reaction. Orthologues of the S. cerevisiae genes encoding the first three enzymes in the arginase pathway were clon...

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Autores principales: Romagnoli, G, Verhoeven, M D, Mans, R, Fleury Rey, Y, Bel-Rhlid, R, van den Broek, M, Maleki Seifar, R, Ten Pierick, A, Thompson, M, Müller, V, Wahl, S A, Pronk, J T, Daran, J M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Blackwell Publishing Ltd 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4149782/
https://www.ncbi.nlm.nih.gov/pubmed/24912400
http://dx.doi.org/10.1111/mmi.12666
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author Romagnoli, G
Verhoeven, M D
Mans, R
Fleury Rey, Y
Bel-Rhlid, R
van den Broek, M
Maleki Seifar, R
Ten Pierick, A
Thompson, M
Müller, V
Wahl, S A
Pronk, J T
Daran, J M
author_facet Romagnoli, G
Verhoeven, M D
Mans, R
Fleury Rey, Y
Bel-Rhlid, R
van den Broek, M
Maleki Seifar, R
Ten Pierick, A
Thompson, M
Müller, V
Wahl, S A
Pronk, J T
Daran, J M
author_sort Romagnoli, G
collection PubMed
description Most available knowledge on fungal arginine metabolism is derived from studies on Saccharomyces cerevisiae, in which arginine catabolism is initiated by releasing urea via the arginase reaction. Orthologues of the S. cerevisiae genes encoding the first three enzymes in the arginase pathway were cloned from Kluyveromyces lactis and shown to functionally complement the corresponding deletion in S. cerevisiae. Surprisingly, deletion of the single K. lactis arginase gene KlCAR1 did not completely abolish growth on arginine as nitrogen source. Growth rate of the deletion mutant strongly increased during serial transfer in shake-flask cultures. A combination of RNAseq-based transcriptome analysis and (13)C-(15)N-based flux analysis was used to elucidate the arginase-independent pathway. Isotopic (13)C(15)N-enrichment in γ-aminobutyrate revealed succinate as the entry point in the TCA cycle of the alternative pathway. Transcript analysis combined with enzyme activity measurements indicated increased expression in the Klcar1Δ mutant of a guanidinobutyrase (EC.3.5.3.7), a key enzyme in a new pathway for arginine degradation. Expression of the K. lactis KLLA0F27995g (renamed KlGBU1) encoding guanidinobutyrase enabled S. cerevisiae to use guanidinobutyrate as sole nitrogen source and its deletion in K. lactis almost completely abolish growth on this nitrogen source. Phylogenetic analysis suggests that this enzyme activity is widespread in fungi.
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spelling pubmed-41497822014-09-17 An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme Romagnoli, G Verhoeven, M D Mans, R Fleury Rey, Y Bel-Rhlid, R van den Broek, M Maleki Seifar, R Ten Pierick, A Thompson, M Müller, V Wahl, S A Pronk, J T Daran, J M Mol Microbiol Research Articles Most available knowledge on fungal arginine metabolism is derived from studies on Saccharomyces cerevisiae, in which arginine catabolism is initiated by releasing urea via the arginase reaction. Orthologues of the S. cerevisiae genes encoding the first three enzymes in the arginase pathway were cloned from Kluyveromyces lactis and shown to functionally complement the corresponding deletion in S. cerevisiae. Surprisingly, deletion of the single K. lactis arginase gene KlCAR1 did not completely abolish growth on arginine as nitrogen source. Growth rate of the deletion mutant strongly increased during serial transfer in shake-flask cultures. A combination of RNAseq-based transcriptome analysis and (13)C-(15)N-based flux analysis was used to elucidate the arginase-independent pathway. Isotopic (13)C(15)N-enrichment in γ-aminobutyrate revealed succinate as the entry point in the TCA cycle of the alternative pathway. Transcript analysis combined with enzyme activity measurements indicated increased expression in the Klcar1Δ mutant of a guanidinobutyrase (EC.3.5.3.7), a key enzyme in a new pathway for arginine degradation. Expression of the K. lactis KLLA0F27995g (renamed KlGBU1) encoding guanidinobutyrase enabled S. cerevisiae to use guanidinobutyrate as sole nitrogen source and its deletion in K. lactis almost completely abolish growth on this nitrogen source. Phylogenetic analysis suggests that this enzyme activity is widespread in fungi. Blackwell Publishing Ltd 2014-07 2014-06-23 /pmc/articles/PMC4149782/ /pubmed/24912400 http://dx.doi.org/10.1111/mmi.12666 Text en © 2014 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd. http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Romagnoli, G
Verhoeven, M D
Mans, R
Fleury Rey, Y
Bel-Rhlid, R
van den Broek, M
Maleki Seifar, R
Ten Pierick, A
Thompson, M
Müller, V
Wahl, S A
Pronk, J T
Daran, J M
An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme
title An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme
title_full An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme
title_fullStr An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme
title_full_unstemmed An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme
title_short An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme
title_sort alternative, arginase-independent pathway for arginine metabolism in kluyveromyces lactis involves guanidinobutyrase as a key enzyme
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4149782/
https://www.ncbi.nlm.nih.gov/pubmed/24912400
http://dx.doi.org/10.1111/mmi.12666
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