Phylogenetic analyses of the condensation and epimerase domains from the <i>Nodularia spumigena</i> CCY9414 aeruginosin gene cluster.

<p>The condensation domain of AerB clusters with the starter condensation domains of nostopeptolide (NosA) and cyanopeptolin (McnA) which load short chain fatty acids and forms a well-supported group. The phylogenetic tree includes all condensation subtypes, including condensation (^LC_L, ^DC_L), condensation and heterocyclization catalyzed by heterocyclization domains, epimerization followed by condensation catalyzed by a Dual E/C domain, and loading condesation domains which are found on initiation modules <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073618#pone.0073618-Rausch1" target="_blank">[29]</a>. The phylogeny was reconstructed using phyml, employing the JTT model of amino acid substitution and a gamma-distributed rate variation with four categories. The support values are based on 100-fold bootstrapping.</p

The identity of the C-terminal fatty acid determined using GC-MS.

<p>Chromatographic behavior of the hexanoic acid standard and the hydrolysed aeruginosin NOL3. Commercial hexanoic acid and hydrolysis product from NOL3 had identical retention times, 4.089 and 4.090 min, respectively, and identical mass spectra which unambiguously prove the first moiety of the NOL3 aeruginosin. (A) hydrolysed aeruginosin NOL3 (B) hexanoic acid standard.</p

Proposed functions of genes in the suspected aeruginosin gene cluster.

<p>The assignment of proposed functions to gene products is based upon BLASTp searches.</p

The production of aeruginosins, spumigins, nodularin, nodulapeptin and suomilide by <i>N. spumigena, N. sphaerocarpa, and N. harveyana</i>.

<p>A maximum-likelihood tree based on the 16S rRNA gene with bootstrap values based on 1000 bootstrap replicates. The taxonomy of the strains follows Lyra et al. 2005 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073618#pone.0073618-Lyra1" target="_blank">[30]</a> and Lehtimäki et al. 2000 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073618#pone.0073618-Lehtimki1" target="_blank">[31]</a>. See also Table S3 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073618#pone.0073618.s001" target="_blank">File S1</a>.</p

Chemical variation of aeruginosins detected from <i>Nodularia spumigena</i>.

<p>Structure, ion mass, retention time and abundance of aeruginosin identified from <i>N. spumigena</i> AV1, CCY9414 and CH307.</p>a<p>Unit mass from ion trap, accurate mass from Q-TOF. Ac = acetic acid, Bu = butyric acid,</p><p>Hex = hexanoic acid, Oct = octanoic acid, Dec = decanoic acid, P = pentose.</p

Location of the spumigin and aeruginosin gene clusters on the circular genome of <i>N. spumigena</i> CCY9414.

<p>The <i>N. spumigena</i> CCY9414 genome encodes 5 non-ribosomal peptide gene clusters. The spumigin (<i>spu</i>) and aeruginosin (<i>aer</i>) gene clusters are encoded 133 kb apart. The spumigin and nodulapeptin (<i>anp</i>) gene clusters are encoded 11 kb apart. The nodularin (<i>nda</i>) gene cluster and a cryptic NRPS gene cluster (unk) are encoded at separate locations on the chromosome.</p

Representatives of the aeruginosin and spumigin families.

<p>These tetrapeptides are produced by strains of <i>Nodularia spumigena</i> isolated from brackish water bodies in Australia and the Baltic Sea. (A) aeruginosin NAL2, (B) an aeruginosin NOL6 containing <i>O</i>-linked pentose, (C) spumigin E.</p