Abstract
The genus Streptomyces represents one of the largest producers of molecules with antibiotic activity. The whole genome sequencing of the endophytic microorganism Streptomyces sp. CBMAI 2042 revealed 35 gene clusters encoding for secondary metabolism including 3 non-ribosomal peptide synthetases (NRPS) and 7 NRPS-hybrids. Combining genome mining and cultivation profile analysis, the depsipeptide ionophore valinomycin was identified as one of the main metabolites produced by this strain. To better understand the metabolic machinery codified in CBMAI 2042 genome an adenylation domain from a hybrid NRPS cluster was deleted through a double crossing knockout experiment. Though the deletion was not plentiful to elucidate the encoded NRP metabolite related to the adenilation domain, an astounding increase of 10.5-fold in valinomycin production was observed in the mutant. These results suggest a metabolic flux redistribution of common substrates as an outcome of a gene target deletion.
Highlights
The genus Streptomyces is a highly prolific producer of structural diverse natural products
biosynthetic gene clusters (BGC) was fully analyzed and correlated with those reported for valinomycin production in Streptomyces tsusimaensis[13] and S. levoris.[15] antiSMASH output of the whole genome sequencing revealed two contiguous open reading frames, spanning 10341 bp to vlm[1] and 8031 bp to vlm[2], both correlated with valinomycin biosynthesis
A deamination reaction to the corresponding keto acids and subsequent reduction produces the hydroxy acid precursors constituents, D-α-hydroxyisovaleric acid (D-Hiv) and L-lactate (L-Lac) from the α-amino acids. These results demonstrate that VLM1 and VLM2 are highly specific for α-ketoisovaleric acid (Kiv) and pyruvate (Pyr) incorporation.[16]
Summary
The genus Streptomyces is a highly prolific producer of structural diverse natural products. The whole genome sequencing of this actinobacteria evidenced a high biosynthetic potential uncovering third five gene clusters related to secondary metabolism. Colonies that did not appear in apramycin-SFM plates were cultivated in TSBY liquid medium and the genomic DNA was extracted for PCR analysis.
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