Arginine and citrulline catabolic pathways encoded by the arc gene cluster of Lactobacillus brevis ATCC 367

Abstract

High concentrations of L-arginine or L-citrulline in the growth medium provided the wine bacterium Lactobacillus brevis with a significant growth advantage. The arginine deiminase pathway (ADI) arc gene cluster of Lactobacillus brevis contains three genes-arcD, arcE1, and arcE2-encoding putative L-arginine/L-ornithine exchangers. Uptake experiments with Lactococcus lactis cells expressing the genes showed that all three transported L-ornithine with affinities in the micromolar range. Similarly, ArcD and ArcE2 transported L-arginine, while ArcE1 transported L-citrulline, an intermediate of the ADI pathway. Chase experiments showed very efficient exchange of L-arginine and L-ornithine by ArcD and ArcE2 and of L-citrulline and L-ornithine by ArcE1. Low affinities (millimolar range) combined with low translocation rates were found for ArcD and ArcE2 with L-citrulline and for ArcE1 with L-arginine. Resting cells of Lactobacillus brevis grown in the presence of L-arginine and L-citrulline rapidly consumed L-arginine and L-citrulline, respectively, while producing ammonia and L-ornithine. About 10% of L-arginine degraded was excreted by the cells as L-citrulline. Degradation of L-arginine and L-citrulline was not subject to carbon catabolite repression by glucose in the medium. At a high medium pH, L-citrulline in the medium was required for induction of the L-citrulline degradation pathway. Pathways are proposed for the catabolic breakdown of L-arginine and L-citrulline that merge at the level of ornithine transcarbamylase in the ADI pathway. L-Arginine uptake is catalyzed by ArcD and/or ArcE2, L-citrulline by ArcE1. L-Citrulline excretion during L-arginine breakdown is proposed to be catalyzed by ArcD and/or ArcE2 through L-arginine/L-citrulline exchange.