TY  - JOUR
A1  - Romagnoli, Gabriele
A1  - Verhoeven, Maarten D.
A1  - Mans, Robert
A1  - Fleury Rey, Yvette
A1  - Bel-Rhlid, Rachid
A1  - Broek, Marcel van den
A1  - Maleki Seifar, Reza
A1  - Pierick, Angela ten
A1  - Thompson, Melanie
A1  - Müller, Volker
A1  - Wahl, Sebastian Aljoscha
A1  - Pronk, Jack T.
A1  - Daran, Jean-Marc
T1  - An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme
T2  - Molecular microbiology
N2  - 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.
Y1  - 2014
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/34313
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-343130
SN  - 1365-2958
SN  - 0950-382X
N1  - © 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.
VL  - 93.2014
IS  - 2
SP  - 369
EP  - 389
ER  -