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Volume 156, Issue 7

Genome-wide transcriptome analyses of the ‘Knallgas’ bacterium H16 with regard to polyhydroxyalkanoate metabolism Free

Abstract

H16 is probably the best-studied ‘Knallgas’ bacterium and producer of poly(3-hydroxybutyrate) (PHB). Genome-wide transcriptome analyses were employed to detect genes that are differentially transcribed during PHB biosynthesis. For this purpose, four transcriptomes from different growth phases of the wild-type H16 and of the two PHB-negative mutants PHB4 and Δ were compared: (i) cells from the exponential growth phase with cells that were in transition to stationary growth phase, and (ii) cells from the transition phase with cells from the stationary growth phase of H16, as well as (iii) cells from the transition phase of H16 with those from the transition phase of PHB4 and (iv) cells from the transition phase of Δ with those from the transition phase of PHB4. Among a large number of genes exhibiting significant changes in transcription level, several genes within the functional class of lipid metabolism were detected. In strain H16, , , , and H16_A3307 exhibited a decreased transcription level in the stationary growth phase compared with the transition phase, whereas , H16_A3311, and were found to be induced in the stationary growth phase. Compared with PHB4, we found that , , H16_A3307, , and were induced in the wild-type, and , , and exhibited an elevated transcription level in PHB4. In strain Δ, and were highly induced compared with PHB4. Additionally, the results of this study suggest that mutant strain PHB4 is defective in PHB biosynthesis and fatty acid metabolism. A significant downregulation of the two operons in mutant strain PHB4 was observed. The putative polyhydroxyalkanoate (PHA) synthase identified in strain H16 was further investigated by several functional analyses. Mutant PHB4 could be phenotypically complemented by expression of from a plasmid; on the other hand, in the mutant H16Δ, no PHA production was observed. PhaC2 activity could not be detected in any experiment.

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Keyword(s): ACP, acyl-carrier proteins , CBB cycle, Calvin–Benson–Bassham cycle , DTNB, 5,5′-dithiobios-(2-nitrobenzoic acid) , GPC, gel permeation chromatography , PHA, polyhydroxyalkanoate and PHB, poly(3-hydroxybutyrate)
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2010-07-01
2024-04-23
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Genome-wide transcriptome analyses of the ‘Knallgas’ bacterium Ralstonia eutropha H16 with regard to polyhydroxyalkanoate metabolism
Microbiology 156, 2136 (2010); https://doi.org/10.1099/mic.0.038380-0
/content/journal/micro/10.1099/mic.0.038380-0
/content/journal/micro/10.1099/mic.0.038380-0
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