Proteins detected in PHA granules of <i>P. extremaustralis</i> by proteome analysis.

<p>The annotated functions of the identified proteins, the locus tags of the coding genes and the calculated molecular masses (MW) are shown.</p

Genetic organization of the mclPHA gene cluster in <i>P. extremaustralis.</i>

<p>mclPHA genes belonging to <i>γ Proteobacteria</i> (smooth arrows). Natural insertion containing 7 ORFs related to <i>β Proteobacteria</i> between the genes <i>phaD</i> and <i>phaF</i> (broken arrows). Arrows indicate the direction of gene transcription and the relative size of each ORF. From left to right: <i>phaC1</i>(1680 bp), <i>phaZ</i> (846 bp), <i>phaC2</i> (1683 bp), <i>phaD</i> (621 bp), followed by genes encoding a LuxR family DNA binding response regulator (642 bp), a putative fimbrial subunit (579 bp), a bacterial pili assembly chaperone (780 bp), a pili assembly chaperone (759 bp), an outer membrane usher protein FimD (2505 bp), a putative fimbrial protein (945 bp) and a Pi fimbriae major subunit (528 bp), not related to PHA metabolism, and <i>phaF</i> (927 bp) and <i>phaI</i> (423 bp).</p

Proteins associated to the PHA granules of <i>P. extremaustralis.</i>

<p>Granules were obtained from cells grown under PHA accumulation conditions. Proteins were separated by SDS-PAGE, stained with Coomassie brilliant blue and subsequently identified by peptide fingerprint analysis as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098873#s2" target="_blank">Materials and Methods</a>. M, molecular mass standard.</p

Bacterial strains and plasmids used in this study.

<p>Bacterial strains and plasmids used in this study.</p

Analysis of the expression of <i>phbC, phaC1</i> and <i>phaC2.</i>

<p>qRT-PCR assays were performed with cells grown on 0.5NE2 supplemented with 0.25% sodium octanoate or 0.27% glucose until late exponential phase. Results are the average ± standard deviations from three independent cultures.</p

Production of PHA in <i>P. extremaustralis</i> and recombinants <i>P. putida</i> GPp104.

<p>Cultures were grown in 0.5NE2 with the indicate carbon source.</p><p>ND =  no detected, PHHX: polyhydroxyhexanoate, PHO: polyhydroxyoctanoate.</p