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Genome, transcriptome and proteome studies indicate that <jats:italic>L. pneumophila<\/jats:italic> also utilizes carbon substrates other than amino acids. We show here that glycerol promotes intracellular replication of <jats:italic>L. pneumophila<\/jats:italic> in amoeba or macrophages (but not extracellular growth) dependent on glycerol\u20103\u2010phosphate dehydrogenase, GlpD. An <jats:italic>L. pneumophila<\/jats:italic> mutant strain lacking <jats:italic>glpD<\/jats:italic> was outcompeted by wild\u2010type bacteria upon co\u2010infection of amoeba, indicating an important role of glycerol during infection. Isotopologue profiling studies using <jats:sup>13<\/jats:sup>C\u2010labelled substrates were performed in a novel minimal defined medium, MDM, comprising essential amino acids, proline and phenylalanine. In MDM, <jats:italic>L. pneumophila<\/jats:italic> utilized <jats:sup>13<\/jats:sup>C\u2010labelled glycerol or glucose predominantly for gluconeogenesis and the pentose phosphate pathway, while the amino acid serine was used for energy generation via the citrate cycle. Similar results were obtained for <jats:italic>L. pneumophila<\/jats:italic> growing intracellularly in amoeba fed with <jats:sup>13<\/jats:sup>C\u2010labelled glycerol, glucose or serine. Collectively, these results reveal a bipartite metabolism of <jats:italic>L. pneumophila<\/jats:italic>, where glycerol and carbohydrates like glucose are mainly fed into anabolic processes, while serine serves as major energy supply.<\/jats:p>","DOI":"10.1111\/mmi.13313","type":"journal-article","created":{"date-parts":[[2015,12,21]],"date-time":"2015-12-21T23:53:09Z","timestamp":1450741989000},"page":"229-246","source":"Crossref","is-referenced-by-count":69,"title":["Pathway analysis using <sup>13<\/sup>C\u2010glycerol and other carbon tracers reveals a bipartite metabolism of <i>Legionella pneumophila<\/i>"],"prefix":"10.1111","volume":"100","author":[{"given":"Ina","family":"H\u00e4uslein","sequence":"first","affiliation":[{"name":"Lehrstuhl f\u00fcr Biochemie, Technische Universit\u00e4t M\u00fcnchen Munich Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Manske","sequence":"additional","affiliation":[{"name":"Max von Pettenkofer 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Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2016,2,19]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2006.03.018"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2002.tb11024.x"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2013.06.012"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2006.00703.x"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijmm.2013.07.003"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.7.3029-3034.1998"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1000851"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng1447"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.2007\/010611-0"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M005508200"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.012363899"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1099776"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.mi.38.100184.000405"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.103.026633"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-11-181"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2010.08.009"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.128678"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2008.06337.x"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2011.00060"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.12450"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2013.06.001"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01043-13"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0052378"},{"key":"e_1_2_6_25_1","first-page":"156","article-title":"Analysis 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