plipastatin-a1 and surfactin-peptide

The self-assembly in aqueous solution of three lipopeptides obtained from Bacillus subtilis has been investigated. The lipopeptides surfactin, plipastatin and mycosubtilin contain distinct cyclic peptide headgroups as well as differences in alkyl chain length, branching and chain length distribution. Cryogenic transmission electron microscopy and X-ray scattering reveal that surfactin and plipastatin aggregate into 2 nm-radius spherical micelles, whereas in complete contrast mycosubtilin self-assembles into extended nanotapes based on bilayer ordering of the lipopeptides. Circular dichroism and FTIR spectroscopy indicate the presence of turn structures in the cyclic peptide headgroup. The unexpected distinct mode of self-assembly of mycosubtilin compared to the other two lipopeptides is ascribed to differences in the surfactant packing parameter. This in turn is due to specific features of the conformation of the peptide headgroup and alkyl chain branching.

Topics: Bacillus subtilis; Cryoelectron Microscopy; Fatty Acids; Lipid Bilayers; Lipopeptides; Lipoproteins; Micelles; Microscopy, Electron, Transmission; Oligopeptides; Peptides, Cyclic; Spectroscopy, Fourier Transform Infrared; X-Ray Diffraction

Microbial competition exists in the general environment, such as soil or aquatic habitats, upon or within unicellular or multicellular eukaryotic life forms. The molecular actions that govern microbial competition, leading to niche establishment and microbial monopolization, remain undetermined. The emerging technology of imaging mass spectrometry (IMS) enabled the observation that there is directionality in the metabolic output of the organism Bacillus subtilis when co-cultured with Staphylococcus aureus. The directionally released antibiotic alters S. aureus virulence factor production and colonization. Therefore, IMS provides insight into the largely hidden nature of competitive microbial encounters and niche establishment, and provides a paradigm for future antibiotic discovery.

Topics: Animals; Bacillus subtilis; Coculture Techniques; Fatty Acids; Female; Humans; Lipopeptides; Male; Mass Spectrometry; Mice; Microbial Viability; Microscopy, Fluorescence; Oligopeptides; Peptides, Cyclic; Staphylococcus aureus

To analyse the effects of plipastatin operon disruption and constitutive expression of surfactin operon in Bacillus subtilis 168 on surfactin productivity, in vitro invasive growth and antagonism against fungi.. The srfA native promoter was replaced by the constitutive promoter P(repU) in B. subtilis 168 after integration of a functional sfp gene. Moreover, the plipastatin synthesis was further disrupted in the B. subtilis 168 derivatives. In liquid media, an earlier and higher expression of P(repU), than that found with P(srfA), led to a specific surfactin production fivefold higher after 6 h of culture. On solid media, not only the invasive growth and the haemolytic activity but also the antifungal activity of the constitutive strains were improved when compared to the parental strain BBG111. As expected, the disruption of the plipastatin operon strongly reduced in vitro antifungal properties but, interestingly, enhanced specific surfactin production (1.47 g g(-1) of biomass), spreading behaviour and haemolytic activity of the strains.. This work demonstrates for the first time the interdependency of surfactin and plipastatin regarding their biosynthesis as well as their influence on the biological activities of the producing strain.. The constitutive overproduction of surfactin enhances the invasive growth and the in vitro antagonistic activity of the mutant strain. Both properties are known to play an important role in the biocontrol of plant diseases. Plipastatin operon disruption increases the surfactin productivity of mutant strains. These mutants are interesting for use in continuous bioprocesses for surfactin production or in bioremediation.

Topics: Bacillus subtilis; Fatty Acids; Fungi; Lipopeptides; Microbial Interactions; Oligopeptides; Operon; Peptides, Cyclic

A biosurfactant-producing strain, Bacillus licheniformis F2.2, was isolated from a fermented food in Thailand. The strain was capable of producing a new biosurfactant, BL1193, as well as two kinds of popular lipopeptide biosurfactants, plipastatin and surfactin. Mass spectrometry and FT-IR analysis indicated that BL1193 had a molecular mass of 1,193 Da with no peptide portion in the molecule. While plipastatin and surfactin were abundantly produced in a nutrient YPD medium, BL1193 was produced only in a synthetic DF medium containing no amino acids. According to an oil displacement activity test, the specific activity of BL1193 (6.53 kBS units/mg) is equivalent to that of surfactin (5.78-6.83 kBS units/mg).

Topics: Bacillus; Fatty Acids; Fermentation; Food Microbiology; Lipopeptides; Lipoproteins; Mass Spectrometry; Oligopeptides; Peptides, Cyclic; Spectroscopy, Fourier Transform Infrared; Surface-Active Agents

Twenty-three halotolerant and biosurfactant producing strains were collected from salty conditions in central Thailand. One of the strains designated BBK-1 produced the biosurfactants with the highest activity. BBK-1 was isolated from fermented foods and was identified as B. subtilis based on its physiological characteristics and 16S rRNA gene sequence. We show that the strain grows in media containing NaCl up to 16% (w/v) and produces biosurfactants in NaCl up to 8%. We found that B. subtilis BBK-1 produces three kinds of surface-active lipopeptides simultaneously. By their respective molecular weights and amino acid compositions, it is indicated that these lipopeptides are bacillomycin L, plipastatin, and surfactin. In order to analyze the production mechanism of lipopeptides further in the strain, a generally important biosynthetic gene encoding 4'-phosphopantetheinyl transferase was cloned and sequenced. The gene existed in a single copy in the genome and the deduced amino acid sequence was almost identical to that of Lpa-14 from B. subtilis strain RB14, which co-produces iturin A and surfactin.

Topics: Amino Acid Sequence; Bacillus subtilis; Bacterial Proteins; Carbon; Cloning, Molecular; DNA, Bacterial; DNA, Ribosomal; Fatty Acids; Fermentation; Food Microbiology; Genes, Bacterial; Lipopeptides; Molecular Sequence Data; Molecular Weight; Oligopeptides; Peptides, Cyclic; RNA, Bacterial; RNA, Ribosomal, 16S; Saline Solution, Hypertonic; Seawater; Sequence Alignment; Sequence Homology, Amino Acid; Soil Microbiology; Species Specificity; Thailand; Transferases (Other Substituted Phosphate Groups); Water Microbiology

Bacillus subtilis YB8 was found to produce the lipopeptide antibiotics surfactin and plipastatin B1. A gene, lpa-8, required for the production of both lipopeptides was cloned from strain YB8. When this gene was inactivated in strain YB8, neither surfactin nor plipastatin B1 was produced. However, the defective strain transformed with an intact lpa-8 gene had restored ability to produce both peptides. Nucleotide sequence analysis of the region essential for the production of the peptides revealed the presence of a large open reading frame. The deduced amino acid sequence of lpa-8 (224 amino acid residues) showed sequence similarity to that of sfp (from surfactin-producing B. subtilis), lpa-14 (from iturin A- and surfactin-producing B. subtilis), psf-1 (from surfactin-producing Bacillus pumilus), gsp (from gramicidin-S-producing Bacillus brevis), and entD (from siderophore-enterobactin-producing Escherichia coli), which are able to complement a defect in the sfp gene and promote production of the lipopeptide antibiotic surfactin. The sequence similarity among these proteins and the product similarity of cyclic peptides suggests that they might be involved in the biosynthesis or secretion of the peptides.

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Antifungal Agents; Bacillus subtilis; Bacterial Proteins; Base Sequence; Cloning, Molecular; DNA, Bacterial; Fatty Acids; Genes, Bacterial; Lipopeptides; Molecular Sequence Data; Molecular Structure; Oligopeptides; Peptides; Peptides, Cyclic; Sequence Homology, Amino Acid; Transformation, Genetic