Isolation and characterization of sfp: a gene that functions in the production of the lipopeptide biosurfactant, surfactin, in Bacillus subtilis

Nakano, Michiko; Corbell, Nathan; Besson, Jerry; Zuber, Peter

doi:10.1007/bf00280011

Cited by 256 publications

(140 citation statements)

References 32 publications

Supporting

Mentioning

126

Contrasting

Unclassified

Order By: Relevance

“…In some cases, such as those of myxothiazol (Silakowski et al, 1999), surfactin (Nakano et al, 1992) or enterobactin (Coderre & Earhart, 1989), the biosynthetic gene clusters code for PPTases. Surprisingly, most gene clusters encoding PKS or NRPS biosynthetic pathways do not contain PPTase genes.…”

Section: Introductionmentioning

confidence: 99%

The phosphopantetheinyl transferase KirP activates the ACP and PCP domains of the kirromycin NRPS/PKS of Streptomyces collinus Tü 365

Pavlidou

Musiol

Kulik

et al. 2011

FEMS Microbiology Letters

View full text Add to dashboard Cite

The main steps in the biosynthesis of complex secondary metabolites such as the antibiotic kirromycin are catalyzed by modular polyketide synthases (PKS) and/or nonribosomal peptide synthetases (NRPS). During antibiotic assembly, the biosynthetic intermediates are attached to carrier protein domains of these megaenzymes via a phosphopantetheinyl arm. This functional group of the carrier proteins is attached post-translationally by a phosphopantetheinyl transferase (PPTase). No experimental evidence exists about how such an activation of the carrier proteins of the kirromycin PKS/NRPS is accomplished. Here we report on the characterization of the PPTase KirP, which is encoded by a gene located in the kirromycin biosynthetic gene cluster. An inactivation of the kirP gene resulted in a 90% decrease in kirromycin production, indicating a substantial role for KirP in the biosynthesis of the antibiotic. In enzymatic assays, KirP was able to activate both acyl carrier protein and petidyl carrier domains of the kirromycin PKS/NRPS. In addition to coenzyme A (CoA), which is the natural substrate of KirP, the enzyme was able to transfer acyl-phosphopantetheinyl groups to the apo forms of the carrier proteins. Thus, KirP is very flexible in terms of both CoA substrate and carrier protein specificity. Our results indicate that KirP is the main PPTases that activates the carrier proteins in kirromycin biosynthesis.

show abstract

Section: Introductionmentioning

confidence: 99%

The phosphopantetheinyl transferase KirP activates the ACP and PCP domains of the kirromycin NRPS/PKS of Streptomyces collinus Tü 365

Pavlidou

Musiol

Kulik

et al. 2011

FEMS Microbiology Letters

View full text Add to dashboard Cite

show abstract

“…Flagella and surfactin are the two most essential gears for B. subtilis swarming (Kearns & Losick, 2003;Julkowska et al, 2004;Kearns et al, 2004). The common laboratory strain of B. subtilis, 168, is defective in surfactin production (Nakano et al, 1992) and hence was reported to be incapable of manifesting swarming motility (Kearns & Losick, 2003). In contrast, the undomesticated or natural wild type B. subtilis strain, 3610, manifests robust swarming (Kearns & Losick, 2003).…”

Section: Introductionmentioning

confidence: 99%

The carboxy terminal domain of Epr, a minor extracellular serine protease, is essential for the swarming motility ofBacillus subtilis168

Murudkar

Kodgire

Rao

2006

FEMS Microbiology Letters

View full text Add to dashboard Cite

In this study we have investigated the role of Epr, a minor extracellular serine protease, in the swarming motility of Bacillus subtilis 168. We identified that the protease activity of Epr was dispensable for swarming. Since the protease activity of Epr was confined to its N-terminal domain, we hypothesized instead that its C-terminal domain (CTD) could be critical for swarming. Our study showed that not only the expression of Epr-CTD was necessary, but also its secretion was crucial for the swarming motility of B. subtilis 168.

show abstract

“…The production of the lipoheptapeptide antibiotic surfactin in Bacillus subtilis depends on the phosphopantetheinyl transferase Sfp (Nakano et al, 1992), which converts the inactive apo-forms of the seven PCP domains of surfactin synthetase (SfrA-ABC) to their active holo-forms (Lambalot et al, 1996). Sfp is likely to play an important role in biotechnology in the future.…”

Section: Introductionmentioning

confidence: 99%

Crystal structure of the surfactin synthetase-activating enzyme Sfp: a prototype of the 4'-phosphopantetheinyl transferase superfamily

et al. 1999

View full text Add to dashboard Cite

The Bacillus subtilis Sfp protein activates the peptidyl carrier protein (PCP) domains of surfactin synthetase by transferring the 4Ј-phosphopantetheinyl moiety of coenzyme A (CoA) to a serine residue conserved in all PCPs. Its wide PCP substrate spectrum renders Sfp a biotechnologically valuable enzyme for use in combinatorial non-ribosomal peptide synthesis. The structure of the Sfp-CoA complex determined at 1.8 Å resolution reveals a novel α/β-fold exhibiting an unexpected intramolecular 2-fold pseudosymmetry. This suggests a similar fold and dimerization mode for the homodimeric phosphopantetheinyl transferases such as acyl carrier protein synthase. The active site of Sfp accommodates a magnesium ion, which is complexed by the CoA pyrophosphate, the side chains of three acidic amino acids and one water molecule. CoA is bound in a fashion that differs in many aspects from all known CoA-protein complex structures. The structure reveals regions likely to be involved in the interaction with the PCP substrate.

show abstract

Isolation and characterization of sfp: a gene that functions in the production of the lipopeptide biosurfactant, surfactin, in Bacillus subtilis

Cited by 256 publications

References 32 publications

The phosphopantetheinyl transferase KirP activates the ACP and PCP domains of the kirromycin NRPS/PKS of Streptomyces collinus Tü 365

The phosphopantetheinyl transferase KirP activates the ACP and PCP domains of the kirromycin NRPS/PKS of Streptomyces collinus Tü 365

The carboxy terminal domain of Epr, a minor extracellular serine protease, is essential for the swarming motility ofBacillus subtilis168

Crystal structure of the surfactin synthetase-activating enzyme Sfp: a prototype of the 4'-phosphopantetheinyl transferase superfamily

Contact Info

Product

Resources

About