title: A lipA (yutB) mutant, encoding lipoic acid synthase, provides insight into the interplay between branched-chain and unsaturated fatty acid biosynthesis in Bacillus subtilis.
abstract: Lipoic acid is an essential cofactor required for the function of key metabolic pathways in most organisms. We report the characterization of a Bacillus subtilis mutant obtained by disruption of the lipA (yutB) gene, which encodes lipoyl synthase (LipA), the enzyme that catalyzes the final step in the de novo biosynthesis of this cofactor. The function of lipA was inferred from the results of genetic and physiological experiments, and this study investigated its role in B. subtilis fatty acid metabolism. Interrupting lipoate-dependent reactions strongly inhibits growth in minimal medium, impairing the generation of branched-chain fatty acids and leading to accumulation of copious amounts of straight-chain saturated fatty acids in B. subtilis membranes. Although depletion of LipA induces the expression of the Delta5 desaturase, controlled by a two-component system that senses changes in membrane properties, the synthesis of unsaturated fatty acids is insufficient to support growth in the absence of precursors for branched-chain fatty acids. However, unsaturated fatty acids generated by deregulated overexpression of the Delta5 desaturase functionally replaces lipoic acid-dependent synthesis of branched-chain fatty acids. Furthermore, we show that the cold-sensitive phenotype of a B. subtilis strain deficient in Delta5 desaturase is suppressed by isoleucine only if LipA is present.
authors.affiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Instituto de Biología Molecular y Celular de Rosario, Universidad Nacional de Rosario, Suipacha 531, 2000 Rosario, Argentina.
authors.forename: Natalia
authors.initials: N
authors.lastname: Martin
authors.forename: Esteban
authors.initials: E
authors.lastname: Lombardía
authors.forename: Silvia G
authors.initials: SG
authors.lastname: Altabe
authors.forename: Diego
authors.initials: D
authors.lastname: de Mendoza
authors.forename: María C
authors.initials: MC
authors.lastname: Mansilla
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D053898:Biosynthetic Pathways; D003470:Culture Media / Q000737:chemistry; D005227:Fatty Acids / Q000096:biosynthesis*; D017353:Gene Deletion; D055786:Gene Knockout Techniques; D013466:Sulfurtransferases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D005227:Fatty Acids; C079999:LipA protein, Bacteria; D013466:Sulfurtransferases; C116435:lipoic acid synthase