Glucose metabolism in the antibiotic producing actinomycete Nonomuraea sp. ATCC 39727

Abstract

The actinomycete Nonomuraea sp. ATCC 39727, producer of the glycopeptide A40926 that is used as precursor for the novel antibiotic dalbavancin, has an unusual carbon metabolism. Glucose is primarily metabolized via the Entner-Doudoroff (ED) pathway, although the energetically more favorable Embden-Meyerhof-Parnas (EMP) pathway is present in this organism. Moreover, Nonomuraea utilizes a PPi-dependent phosphofructokinase, an enzyme that has been connected with anaerobic metabolism in eukaryotes and higher plants, but recently has been recognized in several actinomycetes. In order to study its primary carbon metabolism in further detail, Nonomuraea was cultivated with [1-13C] glucose as the only carbon source and the 13C-labeling patterns of proteinogenic amino acids were determined by GC-MS analysis. Through this method, the fluxes in the central carbon metabolism during balanced growth were estimated. Moreover, a shift in the label incorporation pattern was observed in connection with phosphate limitation and increased antibiotic productivity in Nonomuraea. The shift indicated an increased flux through the EMP pathway at the expense of the flux through the ED pathway, a suggestion that was supported by alterations in intracellular metabolite levels during phosphate limitation. In contrast, expression levels of genes encoding enzymes in the ED and EMP pathways were not affected by phosphate limitation.

Antibiotic Overproduction in Streptomyces coelicolor A3(2) Mediated by Phosphofructokinase Deletion

“…NADPH is the reducing agent used in the process of making secondary metabolites, and the PPP is one of the most important NADPHproducing pathways. A role for NADPH in enhancement of antibiotic production has also been suggested before (1). Another effect could be increased production of acetyl-CoA, which serves as the basic precursor for polyketides, because of high flux through the phosphoketolase pathway.…”

“…Although still largely unclear, the link between primary and secondary metabolism has been repeatedly observed (1)(2)(3). Thus, deletion of the glyceraldehyde 3-phosphate dehydrogenase encoding gene gap1 improved clavulanic acid production in Streptomyces clavuligerus because of increased supply of precursor glyceraldehyde 3-phosphate (4).…”

Integrating multi-omics analyses of Nonomuraea dietziae to reveal the role of soybean oil in [(4′-OH)MeLeu]4-CsA overproduction

“…5, it was worth noting that some key enzymes involved in glycolytic pathway, such as 6-phosphofructokinase (Spot 6, PFK) and pyruvate kinase (Spot 8, PYK) were both present at higher levels. Different from many other actinomycetes, PFK of Nonomuraea is not allosterically regulated by ATP, AMP, ADP, or phosphoenolpyruvate and pyruvate, but controlled by the availability of pyrophosphate (PPi) produced in nucleic acid and protein biosynthesis and in the cycling between glycogen and glucose-1-phosphate [23]. Additionally, the activity of PPi-dependent PFK is reversible, suggesting that N. dietziae had a flexible glycolytic pathway as the regulatory node.…”

Exploring antibiotic biosynthesis: Leo Vining’s insights lead to new strategies in the quest for ‘The 10 × '20 Initiative’

“…However, this is not simple as further complications such as shifts in the metabolic flux during transition from the growth phase to the antibiotic production phase are known to occur. 16 A shift in the precursor supply from growth to antibiotic production makes the interpretation of systems biology data challenging. These complications, along with the fact that precursors can come from several primary metabolic pathways in some cases, make the assessment of the relative importance of individual pathways possible only by comparing the genomes of different strains with different productivity.…”