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Disruption of the siderophore-binding desE receptor gene in Streptomyces coelicolor A3(2) results in impaired growth in spite of multiple iron-siderophore transport systems.

Tierrafría VH, Ramos-Aboites HE, Gosset G, Barona-Gómez F - Microb Biotechnol (2011)

Bottom Line: Here, disruption of the desE gene in S. coelicolor, and subsequent phenotypic analysis, is used to demonstrate a link between iron metabolism and physiological and morphological development.Streptomyces coelicolor desE mutants, isolated in both wild-type (M145) and a coelichelin biosynthesis and transport minus background (mutant W3), a second hydroxamate siderophore system only found in S. coelicolor and related species, resulted in impaired growth and lack of sporulation.The biotechnological and ecological implications of these observations are discussed.

View Article: PubMed Central - PubMed

Affiliation: Evolution of Metabolic Diversity Laboratory, Laboratorio Nacional de Genómica para la Biodiversidad (Langebio), CINVESTAV-IPN, Km 9.6 Libramiento Norte, Carretera Irapuato-León, Irapuato, C.P. 36822, México.

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HPLC and MS analysis of dFOs accumulated by the desE mutant V3. The figure shows a representative chromatogram for M145 and V3, and a mass spectrum for FO‐E (inset). All cultures were grown and treated under identical conditions, i.e. GG1 medium supplemented with 200 µM 2,2′‐dipyridyl and incubation at 30°C for 5 days with vigorous agitation (250 r.p.m.). Two associated peaks, eluting at approximately 16 and 17 min, were identified by MS analysis as FO‐E, whereas FO‐B elutes at 33.6 min, as confirmed with the standard.
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f6: HPLC and MS analysis of dFOs accumulated by the desE mutant V3. The figure shows a representative chromatogram for M145 and V3, and a mass spectrum for FO‐E (inset). All cultures were grown and treated under identical conditions, i.e. GG1 medium supplemented with 200 µM 2,2′‐dipyridyl and incubation at 30°C for 5 days with vigorous agitation (250 r.p.m.). Two associated peaks, eluting at approximately 16 and 17 min, were identified by MS analysis as FO‐E, whereas FO‐B elutes at 33.6 min, as confirmed with the standard.

Mentions: Using the semi‐synthetic liquid medium GG1, which was developed for reproducible production of Streptomyces secondary metabolites (Avignone‐Rossa et al., 2002), the different desE mutants constructed in this study, including their pIJ6902‐derived exconjugants, were analysed by HPLC, and in selected cases the presence of FO‐E and FO‐B on their culture supernatants was confirmed by mass spectrometry (Fig. 6). Again, in order to reproduce an iron‐limited condition, 200 µM 2,2′‐dipyridyl was used. In contrast with the previously Streptomyces iron‐limited liquid medium used to investigate dFOs synthesis as a response to iron deficiency (Barona‐Gómez et al., 2006), this medium reflects more closely industrial conditions, yet due to its defined nature, subsequent analysis of dFOs by HPLC and mass spectrometry is feasible.


Disruption of the siderophore-binding desE receptor gene in Streptomyces coelicolor A3(2) results in impaired growth in spite of multiple iron-siderophore transport systems.

Tierrafría VH, Ramos-Aboites HE, Gosset G, Barona-Gómez F - Microb Biotechnol (2011)

HPLC and MS analysis of dFOs accumulated by the desE mutant V3. The figure shows a representative chromatogram for M145 and V3, and a mass spectrum for FO‐E (inset). All cultures were grown and treated under identical conditions, i.e. GG1 medium supplemented with 200 µM 2,2′‐dipyridyl and incubation at 30°C for 5 days with vigorous agitation (250 r.p.m.). Two associated peaks, eluting at approximately 16 and 17 min, were identified by MS analysis as FO‐E, whereas FO‐B elutes at 33.6 min, as confirmed with the standard.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3818867&req=5

f6: HPLC and MS analysis of dFOs accumulated by the desE mutant V3. The figure shows a representative chromatogram for M145 and V3, and a mass spectrum for FO‐E (inset). All cultures were grown and treated under identical conditions, i.e. GG1 medium supplemented with 200 µM 2,2′‐dipyridyl and incubation at 30°C for 5 days with vigorous agitation (250 r.p.m.). Two associated peaks, eluting at approximately 16 and 17 min, were identified by MS analysis as FO‐E, whereas FO‐B elutes at 33.6 min, as confirmed with the standard.
Mentions: Using the semi‐synthetic liquid medium GG1, which was developed for reproducible production of Streptomyces secondary metabolites (Avignone‐Rossa et al., 2002), the different desE mutants constructed in this study, including their pIJ6902‐derived exconjugants, were analysed by HPLC, and in selected cases the presence of FO‐E and FO‐B on their culture supernatants was confirmed by mass spectrometry (Fig. 6). Again, in order to reproduce an iron‐limited condition, 200 µM 2,2′‐dipyridyl was used. In contrast with the previously Streptomyces iron‐limited liquid medium used to investigate dFOs synthesis as a response to iron deficiency (Barona‐Gómez et al., 2006), this medium reflects more closely industrial conditions, yet due to its defined nature, subsequent analysis of dFOs by HPLC and mass spectrometry is feasible.

Bottom Line: Here, disruption of the desE gene in S. coelicolor, and subsequent phenotypic analysis, is used to demonstrate a link between iron metabolism and physiological and morphological development.Streptomyces coelicolor desE mutants, isolated in both wild-type (M145) and a coelichelin biosynthesis and transport minus background (mutant W3), a second hydroxamate siderophore system only found in S. coelicolor and related species, resulted in impaired growth and lack of sporulation.The biotechnological and ecological implications of these observations are discussed.

View Article: PubMed Central - PubMed

Affiliation: Evolution of Metabolic Diversity Laboratory, Laboratorio Nacional de Genómica para la Biodiversidad (Langebio), CINVESTAV-IPN, Km 9.6 Libramiento Norte, Carretera Irapuato-León, Irapuato, C.P. 36822, México.

Show MeSH
Related in: MedlinePlus