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

Romagnoli G, Verhoeven MD, Mans R, Rey YF, Bel-Rhlid R, Van Den Broek M, Seifar RM, Ten Pierick A, Thompson M, Mueller V, Pronk JT, Daran JM, Wahl SA (2014)

Publication Type: Journal article

Publication year: 2014

Journal

Molecular Microbiology Wiley-Blackwell

Book Volume: 93

Pages Range: 369-389

Journal Issue: 2

DOI: 10.1111/mmi.12666

Abstract

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. lactisKLLA0F27995g (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. © 2014 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.

Authors with CRIS profile

Sebastian Aljoscha Wahl

Involved external institutions

Delft University of Technology (TU Delft) NL Netherlands (NL) Goethe-Universität Frankfurt am Main DE Germany (DE) Nestlé S.A. CH Switzerland (CH)

How to cite

APA:

Romagnoli, G., Verhoeven, M.D., Mans, R., Rey, Y.F., Bel-Rhlid, R., Van Den Broek, M.,... Wahl, S.A. (2014). An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme. Molecular Microbiology, 93(2), 369-389. https://dx.doi.org/10.1111/mmi.12666

MLA:

Romagnoli, G., et al. "An alternative, arginase-independent pathway for arginine metabolism in Kluyveromyces lactis involves guanidinobutyrase as a key enzyme." Molecular Microbiology 93.2 (2014): 369-389.

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