Dataset Information

Lactobacillus delbrueckii subspecies characterization by MudPIT analysis

ABSTRACT: We have characterized three strains belonging to L. delbrueckii subspecies (bulgaricus, B; lactis, L; L.delbrueckii 23, M) highlighting their divergent origin, different galactose metabolic capabilities and we have demonstrated potential probiotic features.

INSTRUMENT(S): LTQ

ORGANISM(S): Lactobacillus Delbrueckii

SUBMITTER: Sara Motta

LAB HEAD: Pierluigi Mauri

PROVIDER: PXD006551 | Pride | 2017-06-29

REPOSITORIES: Pride

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Combination of Metabolomic and Proteomic Analysis Revealed Different Features among Lactobacillus delbrueckii Subspecies bulgaricus and lactis Strains While In Vivo Testing in the Model Organism Caenorhabditis elegans Highlighted Probiotic Properties.

Zanni Elena E Schifano Emily E Motta Sara S Sciubba Fabio F Palleschi Claudio C Mauri Pierluigi P Perozzi Giuditta G Uccelletti Daniela D Devirgiliis Chiara C Miccheli Alfredo A

Frontiers in microbiology 20170628

Lactobacillus delbrueckii represents a technologically relevant member of lactic acid bacteria, since the two subspecies bulgaricus and lactis are widely associated with fermented dairy products. In the present work, we report the characterization of two commercial strains belonging to L. delbrueckii subspecies bulgaricus, lactis and a novel strain previously isolated from a traditional fermented fresh cheese. A phenomic approach was performed by combini ...[more]

PMID: 28702021

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Project 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. Orthologs 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 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 ?-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 Klcar1? mutant of a guanidinobutyrase (EC.3.5.3.7), an enzyme not previously demonstrated in fungi. 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. The goal of the present study was to characterize arginine catabolism in K. lactis. To this end, CAR1, CAR2 and PRO3 orthologs in K. lactis were identified and functionally analysed by deletion, expression in S. cerevisiae and enzyme activity assays. Since deletion of the arginase gene in K. lactis was found not to completely abolish growth on arginine as a sole nitrogen source, the alternative pathway for arginine catabolism operating in this yeast was studied by a combination of transcriptome analysis, 13C and 15N isotope-based flux analysis and enzyme activity assays in cell extracts. To investigate arginine metabolism in the arginase-negative K. lactis strain, strains GG1632 (Klku80? KlCAR1 reference strain) and IMS0367 (Klcar1? Arg+) were grown in aerobic bioreactor batch cultures on glucose chemically defined medium with arginine as sole nitrogen source. RNA sequencing of samples taken during the exponential phase of growth on glucose-arginine media of the reference strain G1631 and the arginase less strain IMS0367 were compared resulting in the characterization of a new function.

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Dataset Information

Lactobacillus delbrueckii subspecies characterization by MudPIT analysis

Dataset's files

Publications

Combination of Metabolomic and Proteomic Analysis Revealed Different Features among <i>Lactobacillus delbrueckii</i> Subspecies <i>bulgaricus</i> and <i>lactis</i> Strains While <i>In Vivo</i> Testing in the Model Organism <i>Caenorhabditis elegans</i> Highlighted Probiotic Properties.

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