Microparticles globally reprogram the metabolism of Streptomyces albus
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA648406 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR12318586_1.fastq.gz | Fastqsanger.gz | |||
| SRR12318586_2.fastq.gz | Fastqsanger.gz | |||
| SRR12318587_1.fastq.gz | Fastqsanger.gz | |||
| SRR12318587_2.fastq.gz | Fastqsanger.gz | |||
| SRR12318588_1.fastq.gz | Fastqsanger.gz |
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Project description:Streptomyces sp. are a rich source for natural products with recognized industrial value, explaining the high interest to improve and streamline production in these microbes. Here, we studied the production of pamamycins, macrodiolide homologues with a high activity against multi-resistant pathogenic microbes, using recombinant S. albus J1074/R2. Talc particles of micrometer size added to submerged cultures of the recombinant strain tripled pamamycin production up to 50 mg L-1. Furthermore, they strongly affected morphology, reduced the size of cell pellets, formed by the filamentous microbe during the process, up to six-fold, and shifted the pamamycin spectrum to larger derivatives. Integrated analysis of transcriptome and metabolome of particle-enhanced and control cultures provided detailed insights into the underlying molecular changes. The microparticles affected the expression of 3341 genes (56%), revealing a global and fundamental impact on metabolism. Morphology-associated genes, encoding major regulators such as SsgA, RelA, EshA, Factor C, as well as chaplins and rodlins, were found massively upregulated, indicating that the particles caused a substantially accelerated morphogenesis. In line, the pamamycin cluster was strongly upregulated (up to log2 10-fold). Furthermore, the microparticles perturbed genes encoding for central catabolism and CoA-ester metabolism, which were mainly activated. The altered expression resulted in changes in the availability of intracellular CoA-esters, the building blocks of pamamycin. Notably, the ratio between methylmalonyl CoA and malonyl-CoA was increased four-fold. Both metabolites compete for incorporation into pamamycin so that the altered availability explained the pronounced preference for larger derivatives in the microparticle-enhanced process. Our findings are straightforward to further develop pamamycins into antituberculosis leads. The novel insights into the behavior of S. albus in response to talc appears of general relevance to further explore and upgrade the concept of microparticle enhanced cultivation, widely used for filamentous microbes.
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Project description:Despite their essential role in fueling the onset of metabolism, the composition and functional state of seed mitochondria are still a matter of debate. Using white lupin (Lupinus albus) as a model, we provide a comprehensive proteomic characterization of seed mitochondria in a legume with protein-rich reserves. Highly enriched mitochondrial fractions isolated from quiescent seeds revealed fully assembled oxidative phosphorylation (OXPHOS) complexes and supercomplexes, demonstrating that mitochondria are preconfigured for immediate energy production upon imbibition. Quantitative proteomics identified 1,162 mitochondrial proteins and highlighted marked tissue-specific differences compared to leaves, including distinct transporter profiles and a dehydrogenase complement supporting amino acid and fatty acid catabolism. Early germination was accompanied by remodeling of coenzyme metabolism and transport capacity, while core respiratory complexes remained stable. Notably, ~12% of the proteome consisted of uncharacterized proteins, many of which displayed dynamic changes during early germination, suggesting yet undiscovered, potentially legume-specific mitochondrial functions.
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Project description:To evaluate the DNA binding of AntA in Streptomyces albus S4, in vivo ChIP-Seq experiments were performed 3xFLAG-AntA and anti-FLAG antibodies using two biological replicates. The wild type strain was used as a control in experiments using the anti-FLAG antibodies as well as a DNA only control (which were published previously, E-MTAB-5122).
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