title: Differential proteomic analysis reveals novel links between primary metabolism and antibiotic production in Amycolatopsis balhimycina.
abstract: A differential proteomic analysis, based on 2-DE and MS procedures, was performed on Amycolatopsis balhimycina DSM5908, the actinomycete producing the vancomycin-like antibiotic balhimycin. A comparison of proteomic profiles before and during balhimycin production characterized differentially and constitutively expressed protein isoforms, which were associated with 203 ORFs in the A. balhimycina genome. These data, providing insights on the major metabolic pathways/molecular processes operating in this organism, were used to compile 2-DE reference maps covering 3-10, 4-7 and 4.5-5.5 pH gradients available over the World Wide Web as interactive web pages (http://www.unipa.it/ampuglia/Abal-proteome-maps). Functional clustering analysis revealed that differentially expressed proteins belong to functional groups involved in central carbon metabolism, amino acid metabolism and protein biosynthesis, energetic and redox balance, sugar/amino sugar metabolism, balhimycin biosynthesis and transcriptional regulation or with hypothetical and/or unknown function. Interestingly, proteins involved in the biosynthesis of balhimycin precursors, such as amino acids, amino sugars and central carbon metabolism intermediates, were upregulated during antibiotic production. qRT-PCR analysis revealed that 8 out of 14 upregulated genes showed a positive correlation between changes at translational and transcriptional expression level. Furthermore, proteomic analysis of two nonproducing mutants, restricted to a sub-set of differentially expressed proteins, showed that most proteins required for the biosynthesis of balhimycin precursors are downregulated in both mutants. These findings suggest that primary metabolic pathways support anabolic routes leading to balhimycin biosynthesis and the differentially expressed genes are interesting targets for the construction of high-yielding producer strains by rational genetic engineering.
authors.affiliation: Dipartimento di Biologia Cellulare e dello Sviluppo, University of Palermo, Palermo, Italy. giuseppe.gallo@unipa.it
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gallo
authors.forename: Giovanni
authors.initials: G
authors.lastname: Renzone
authors.forename: Rosa
authors.initials: R
authors.lastname: Alduina
authors.forename: Efthimia
authors.initials: E
authors.lastname: Stegmann
authors.forename: Tilmann
authors.initials: T
authors.lastname: Weber
authors.forename: Anna Eliasson
authors.initials: AE
authors.lastname: Lantz
authors.forename: Jette
authors.initials: J
authors.lastname: Thykaer
authors.forename: Fabio
authors.initials: F
authors.lastname: Sangiorgi
authors.forename: Andrea
authors.initials: A
authors.lastname: Scaloni
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Puglia
mesh_terms: D000192:Actinomycetales / Q000235:genetics / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D016000:Cluster Analysis; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry / Q000379:methods; D053858:Metabolic Networks and Pathways; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D014640:Vancomycin / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D005656:Fungal Proteins; D020543:Proteome; C087127:balhimycin; D014640:Vancomycin