title: Genetic engineering of the branched fatty acid metabolic pathway of Bacillus subtilis for the overproduction of surfactin C14isoform.
abstract: Surfactin, a lipopeptide produced by Bacillus subtilis, is one of the most powerful biosurfactants known. This molecule consists of a cyclic heptapeptide linked to a β-hydroxy fatty acid chain. The isomery and the length of the fatty acid (FA) chain are responsible for the surfactin's activities. In this study, the gene codY, which encode for the global transcriptional regulator and the gene lpdV, located in the bkd operon (lpdV, bkdAA, bkdAB and bkdB genes), which is responsible for the last step of the branched chain amino acid (BCAA) degradation in acyl-CoA were deleted. The influence of these deletions on the quantitative and qualitative surfactin production was analysed. The surfactin production was quantified by RP-HPLC and the surfactin isoforms were characterized using LC-MS-MS and GC-MS analysis. The results obtained in the mutants showed an enhancement of surfactin specific production by a factor of 5.8 for the codY mutant and 1.4 for lpdV mutant. Moreover qualitative analysis of the lpdV mutant reveals that it mainly produced surfactin C14isoform (2 fold more than the wild type) with linear FA chain. Complete analysis of the extracellular metabolites using1H quantitative NMR reveals a reduced production of acetoin in this mutant. This work demonstrates for the first time an original approach to overproduce specifically surfactin with C14FA chain.
authors.affiliation: University Lille, INRA, ISA, University Artois, University Littoral Côte d'Opale, EA 7394 - ICV - Institut Charles Viollette, Lille, France.
authors.forename: Debarun
authors.initials: D
authors.lastname: Dhali
authors.affiliation: University Lille, INRA, ISA, University Artois, University Littoral Côte d'Opale, EA 7394 - ICV - Institut Charles Viollette, Lille, France.
authors.forename: François
authors.initials: F
authors.lastname: Coutte
authors.affiliation: MiPI, TERRA Research Centre, Gembloux Agro-Bio Tech, University of Liege, Passage des Déportés, Gembloux, Belgium.
authors.forename: Anthony Argüelles
authors.initials: AA
authors.lastname: Arias
authors.affiliation: Micalis Institute, INRA, AgroParisTech, University Paris-Saclay, Jouy-en-Josas, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Auger
authors.affiliation: Micalis Institute, INRA, AgroParisTech, University Paris-Saclay, Jouy-en-Josas, France.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Bidnenko
authors.affiliation: University Lille, INRA, ISA, University Artois, University Littoral Côte d'Opale, EA 7394 - ICV - Institut Charles Viollette, Lille, France.
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Chataigné
authors.affiliation: Ernst-Moritz-Arndt-University Greifswald, Institute of Biochemistry, Greifswald, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Lalk
authors.affiliation: University Lille, BioComputing Team, CRIStAL Lab (CNRS UMR9189), Villeneuve d'Ascq, France.|Inria Lille, Villeneuve d'Ascq, France.
authors.forename: Joachim
authors.initials: J
authors.lastname: Niehren
authors.affiliation: Ernst-Moritz-Arndt-University Greifswald, Institute of Biochemistry, Greifswald, Germany.
authors.forename: Joana
authors.initials: J
authors.lastname: de Sousa
authors.affiliation: University Lille, BioComputing Team, CRIStAL Lab (CNRS UMR9189), Villeneuve d'Ascq, France.
authors.forename: Cristian
authors.initials: C
authors.lastname: Versari
authors.affiliation: University Lille, INRA, ISA, University Artois, University Littoral Côte d'Opale, EA 7394 - ICV - Institut Charles Viollette, Lille, France.|MiPI, TERRA Research Centre, Gembloux Agro-Bio Tech, University of Liege, Passage des Déportés, Gembloux, Belgium.
authors.forename: Philippe
authors.initials: P
authors.lastname: Jacques
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics / Q000254:growth & development*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002853:Chromatography, Liquid; D005227:Fatty Acids / Q000096:biosynthesis*; D008401:Gas Chromatography-Mass Spectrometry; D017353:Gene Deletion; D005818:Genetic Engineering; D055666:Lipopeptides / Q000235:genetics / Q000378:metabolism*; D053858:Metabolic Networks and Pathways; D009876:Operon; D020033:Protein Isoforms / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D001426:Bacterial Proteins; D005227:Fatty Acids; D055666:Lipopeptides; D020033:Protein Isoforms
keywords: Bacillus subtilis; Fatty acid metabolic pathway; Lipopeptide; Surfactin