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 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 13C-15N-based flux analysis was used to elucidate the arginase-independent pathway. Isotopic 13C15N-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.

Download full text files

Export metadata

Additional Services

Share in Twitter Search Google Scholar
Metadaten
Author:Gabriele Romagnoli, Maarten D. Verhoeven, Robert Mans, Yvette Fleury Rey, Rachid Bel-Rhlid, Marcel van den Broek, Reza Maleki Seifar, Angela ten Pierick, Melanie Thompson, Volker MüllerORCiD, Sebastian Aljoscha Wahl, Jack T. PronkORCiDGND, Jean-Marc Daran
URN:urn:nbn:de:hebis:30:3-343130
DOI:https://doi.org/10.1111/mmi.12666
ISSN:1365-2958
ISSN:0950-382X
Pubmed Id:https://pubmed.ncbi.nlm.nih.gov/24912400
Parent Title (German):Molecular microbiology
Document Type:Article
Language:English
Date of Publication (online):2016/11/21
Date of first Publication:2014/06/23
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2016/11/21
Volume:93.2014
Issue:2
Page Number:21
First Page:369
Last Page:389
Note:
© 2014 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd. 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.
HeBIS-PPN:428652891
Institutes:Biowissenschaften / Biowissenschaften
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Licence (German):License LogoCreative Commons - Namensnennung-Nicht kommerziell - Keine Bearbeitung 4.0