title: Utilization of D-ribitol by Lactobacillus casei BL23 requires a mannose-type phosphotransferase system and three catabolic enzymes.
abstract: Lactobacillus casei strains 64H and BL23, but not ATCC 334, are able to ferment D-ribitol (also called D-adonitol). However, a BL23-derived ptsI mutant lacking enzyme I of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) was not able to utilize this pentitol, suggesting that strain BL23 transports and phosphorylates D-ribitol via a PTS. We identified an 11-kb region in the genome sequence of L. casei strain BL23 (LCABL_29160 to LCABL_29270) which is absent from strain ATCC 334 and which contains the genes for a GlpR/IolR-like repressor, the four components of a mannose-type PTS, and six metabolic enzymes potentially involved in D-ribitol metabolism. Deletion of the gene encoding the EIIB component of the presumed ribitol PTS indeed prevented D-ribitol fermentation. In addition, we overexpressed the six catabolic genes, purified the encoded enzymes, and determined the activities of four of them. They encode a D-ribitol-5-phosphate (D-ribitol-5-P) 2-dehydrogenase, a D-ribulose-5-P 3-epimerase, a D-ribose-5-P isomerase, and a D-xylulose-5-P phosphoketolase. In the first catabolic step, the protein D-ribitol-5-P 2-dehydrogenase uses NAD(+) to oxidize D-ribitol-5-P formed during PTS-catalyzed transport to D-ribulose-5-P, which, in turn, is converted to D-xylulose-5-P by the enzyme D-ribulose-5-P 3-epimerase. Finally, the resulting D-xylulose-5-P is split by D-xylulose-5-P phosphoketolase in an inorganic phosphate-requiring reaction into acetylphosphate and the glycolytic intermediate D-glyceraldehyde-3-P. The three remaining enzymes, one of which was identified as D-ribose-5-P-isomerase, probably catalyze an alternative ribitol degradation pathway, which might be functional in L. casei strain 64H but not in BL23, because one of the BL23 genes carries a frameshift mutation.
authors.affiliation: INRA, Microbiologie de l'alimentation au service de la santé humaine (MICALIS), UMR1319, Jouy en Josas, France.
authors.forename: A
authors.initials: A
authors.lastname: Bourand
authors.forename: M J
authors.initials: MJ
authors.lastname: Yebra
authors.forename: G
authors.initials: G
authors.lastname: Boël
authors.forename: A
authors.initials: A
authors.lastname: Mazé
authors.forename: J
authors.initials: J
authors.lastname: Deutscher
mesh_terms: D000446:Aldehyde-Lyases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D002238:Carbohydrate Epimerases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D005285:Fermentation; D015870:Gene Expression; D007780:Lacticaseibacillus casei / Q000201:enzymology* / Q000235:genetics; D008358:Mannose / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D008969:Molecular Sequence Data; D009154:Mutation; D009243:NAD / Q000378:metabolism; D009876:Operon; D010428:Pentosephosphates / Q000378:metabolism; D010728:Phosphoenolpyruvate / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012255:Ribitol / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D010428:Pentosephosphates; D011993:Recombinant Fusion Proteins; D009243:NAD; D012255:Ribitol; C031625:xylulose-5-phosphate; D010728:Phosphoenolpyruvate; D013401:Sugar Alcohol Dehydrogenases; C044691:ribitol-5-phosphate dehydrogenase; D000446:Aldehyde-Lyases; C022367:phosphoketolase; D002238:Carbohydrate Epimerases; C022553:ribulosephosphate 3-epimerase; D008358:Mannose
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