title: Ca2+-citrate uptake and metabolism in Lactobacillus casei ATCC 334.
abstract: The putative citrate metabolic pathway in Lactobacillus casei ATCC 334 consists of the transporter CitH, a proton symporter of the citrate-divalent metal ion family of transporters CitMHS, citrate lyase, and the membrane-bound oxaloacetate decarboxylase complex OAD-ABDH. Resting cells of Lactobacillus casei ATCC 334 metabolized citrate in complex with Ca(2+) and not as free citrate or the Mg(2+)-citrate complex, thereby identifying Ca(2+)-citrate as the substrate of the transporter CitH. The pathway was induced in the presence of Ca(2+) and citrate during growth and repressed by the presence of glucose and of galactose, most likely by a carbon catabolite repression mechanism. The end products of Ca(2+)-citrate metabolism by resting cells of Lb. casei were pyruvate, acetate, and acetoin, demonstrating the activity of the membrane-bound oxaloacetate decarboxylase complex OAD-ABDH. Following pyruvate, the pathway splits into two branches. One branch is the classical citrate fermentation pathway producing acetoin by α-acetolactate synthase and α-acetolactate decarboxylase. The other branch yields acetate, for which the route is still obscure. Ca(2+)-citrate metabolism in a modified MRS medium lacking a carbohydrate did not significantly affect the growth characteristics, and generation of metabolic energy in the form of proton motive force (PMF) was not observed in resting cells. In contrast, carbohydrate/Ca(2+)-citrate cometabolism resulted in a higher biomass yield in batch culture. However, also with these cells, no generation of PMF was associated with Ca(2+)-citrate metabolism. It is concluded that citrate metabolism in Lb. casei is beneficial when it counteracts acidification by carbohydrate metabolism in later growth stages.
authors.affiliation: Molecular Microbiology, Groningen Biomolecular Science and Biotechnology Institute, University of Groningen, Groningen, Netherlands.
authors.forename: Pablo
authors.initials: P
authors.lastname: Mortera
authors.forename: Agata
authors.initials: A
authors.lastname: Pudlik
authors.forename: Christian
authors.initials: C
authors.lastname: Magni
authors.forename: Sergio
authors.initials: S
authors.lastname: Alarcón
authors.forename: Juke S
authors.initials: JS
authors.lastname: Lolkema
mesh_terms: D019355:Calcium Citrate / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D002262:Carboxy-Lyases / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002952:Citric Acid Cycle; D003124:Colorimetry; D004734:Energy Metabolism; D005285:Fermentation; D007780:Lacticaseibacillus casei / Q000254:growth & development / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D002262:Carboxy-Lyases; C023746:acetolactate decarboxylase; D019355:Calcium Citrate