A Polyketide Synthase in Glycopeptide Biosynthesis

Pfeifer, Volker; Nicholson, Graeme; Ries, Johannes B.; Recktenwald, Jürgen; Schefer, Alexandre B.; Shawky, Riham M.; Schröder, Joachim; Wohlleben, Wolfgang; Pelzer, Stefan

doi:10.1074/jbc.m106580200

Cited by 141 publications

(104 citation statements)

References 33 publications

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“…4E) that the A-PCP domains of the seemingly superfluous GetM protein are active, this di-domain may play a role in the biosynthesis of the fourth amino acid precursor. Indeed, there are a number of examples in which specialized A-PCP di-domains are essential for generating NRPS precursors (52)(53)(54)(55)(56). Here, we propose that the A-domain GetMA 5 activates (2S)-histidine, consistent with the results of the ATP-PP i exchange assay, and tethers it to the PCP of GetM.…”

Section: Discussionsupporting

confidence: 73%

Insights into an Unusual Nonribosomal Peptide Synthetase Biosynthesis

Binz¹,

Maffioli²,

Sosio³

et al. 2010

Journal of Biological Chemistry

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Section: Discussionsupporting

confidence: 73%

Insights into an Unusual Nonribosomal Peptide Synthetase Biosynthesis

Binz¹,

Maffioli²,

Sosio³

et al. 2010

Journal of Biological Chemistry

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“…DpgA is a polyketide synthase that generates 3,5-dihydroxyphenylacetyl-CoA from four molecules of malonyl-CoA, with the assistance of DpgB and DpgD, proteins that exhibit dehydratase activity. The oxidase DpgC converts 3,5-dihydroxyphenylacetyl-CoA to 3,5-dihydroxyphenylglyoxylate, which generates DHPG on transamination by HpgT (21). Orthologous genes encoding these enzymes in S. toyocaensis, ORFs 12-15, are found arranged as they are in other glycopeptide clusters (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…The S. toyocaensis orthologous genes are grouped in a similar fashion to those in the complestatin biosynthetic cluster in contrast with the equivalent chloroeremomycin genes, which are distributed throughout the cluster. The biochemical details of DHPG biosynthesis have been investigated in the chloroeremomcyin producer A. orientalis, and involve four characterized enzymes and one additional transamination step (21,22). DpgA is a polyketide synthase that generates 3,5-dihydroxyphenylacetyl-CoA from four molecules of malonyl-CoA, with the assistance of DpgB and DpgD, proteins that exhibit dehydratase activity.…”

Section: Resultsmentioning

confidence: 99%

Assembling the glycopeptide antibiotic scaffold: The biosynthesis of from Streptomyces toyocaensis NRRL15009

Pootoolal

Thomas

Marshall

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

171

163

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The glycopeptide antibiotics vancomycin and teicoplanin are vital components of modern anti-infective chemotherapy exhibiting outstanding activity against Gram-positive pathogens including members of the genera Streptococcus, Staphylococcus, and Enterococcus. These antibiotics also provide fascinating examples of the chemical and associated biosynthetic complexity exploitable in the synthesis of natural products by actinomycetes group of bacteria. We report the sequencing and annotation of the biosynthetic gene cluster for the glycopeptide antibiotic A47934 from Streptomyces toyocaensis NRRL15009, the first complete sequence for a teicoplanin class glycopeptide. The cluster includes 34 ORFs encompassing 68 kb and includes all of the genes predicted to be required to synthesize A47934 and regulate its biosynthesis. The gene cluster also contains ORFs encoding enzymes responsible for glycopeptide resistance. This role was confirmed by insertional inactivation of the D-Ala-D-lactate ligase, vanAst, which resulted in the predicted A47934-sensitive phenotype and impaired antibiotic biosynthesis. These results provide increased understanding of the biosynthesis of these complex natural products.G lycopeptide antibiotics (GPAs) have been mainstays of antimicrobial chemotherapy since their discovery in the mid 1950s. These antibiotics act exclusively on Gram-positive bacteria by forming a tight and specific noncovalent complex with the D-Ala-D-Ala terminus of the peptidoglycan, inhibiting cell wall growth and crosslinking (1, 2). Clinical resistance to GPAs was first described in the enterococci in 1988 (3), and resistance has now manifested itself in the more virulent streptococci and staphylococci (reviewed in ref. 4). GPA resistance is now a significant worldwide phenomenon that has severely impacted the health care sector both in increased mortality and morbidity, and economically (5). The predominant mechanism of resistance is the synthesis of cell wall peptidoglycan terminating in D-Ala-D-lactate, which dramatically decreases the affinity of these antibiotics for their target (6).GPAs are exclusively obtained through fermentation, yet despite their importance, their biosynthesis is not well understood. They are comprised of a heptapeptide core consisting of both common and unusual amino acids (4). Crosslinking of the amino acids through aryl ether and carbon-carbon bonds provides rigidity to the peptide. There are two major structural classes of GPAs based on the identity of the core peptide, and the two clinically used GPAs, vancomycin and teicoplanin, exemplify both classes (Fig. 1). An additional class of structurally homologous secondary metabolites with anticomplement activity is exemplified by complestatin (Fig. 1). Structural diversity in these natural products is achieved through changes in the peptide backbone, and selective amino acid halogenation, glycosylation, lipidation, methylation, and sulfonylation. In principle, understanding of the genetic and mechanistic basis of these modifications could lead ...

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“…strain ATCC 39727 as a complex of related compounds. It consists of a heptapeptide containing the proteinogenic amino acid tyrosine and the nonproteinogenic amino acids 3,5-dihydroxyphenylglycine (DPG) and 4-hydroxyphenylglycine (HPG) (1)(2)(3)(4). The heptapeptide is assembled by a nonribosomal peptide synthetase (NRPS) and then modified by oxidative cross-linking of the aromatic side chains to yield a rigid peptide scaffold.…”

mentioning

confidence: 99%

Two Master Switch Regulators Trigger A40926 Biosynthesis in Nonomuraea sp. Strain ATCC 39727

Grasso

Maffioli²,

Sosio³

et al. 2015

J Bacteriol

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The actinomycete Nonomuraea sp. strain ATCC 39727 produces the glycopeptide A40926, the precursor of dalbavancin. Biosynthesis of A40926 is encoded by the dbv gene cluster, which contains 37 protein-coding sequences that participate in antibiotic biosynthesis, regulation, immunity, and export. In addition to the positive regulatory protein Dbv4, the A40926-biosynthetic gene cluster encodes two additional putative regulators, Dbv3 and Dbv6. Independent mutations in these genes, combined with bioassays and liquid chromatography-mass spectrometry (LC-MS) analyses, demonstrated that Dbv3 and Dbv4 are both required for antibiotic production, while inactivation of dbv6 had no effect. In addition, overexpression of dbv3 led to higher levels of A40926 production. Transcriptional and quantitative reverse transcription (RT)-PCR analyses showed that Dbv4 is essential for the transcription of two operons, dbv14-dbv8 and dbv30-dbv35, while Dbv3 positively controls the expression of four monocistronic transcription units (dbv4, dbv29, dbv36, and dbv37) and of six operons (dbv2-dbv1, dbv14-dbv8, dbv17-dbv15, dbv21-dbv20, dbv24-dbv28, and dbv30-dbv35). We propose a complex and coordinated model of regulation in which Dbv3 directly or indirectly activates transcription of dbv4 and controls biosynthesis of 4-hydroxyphenylglycine and the heptapeptide backbone, A40926 export, and some tailoring reactions (mannosylation and hexose oxidation), while Dbv4 directly regulates biosynthesis of 3,5-dihydroxyphenylglycine and other tailoring reactions, including the four cross-links, halogenation, glycosylation, and acylation. IMPORTANCEThis report expands knowledge of the regulatory mechanisms used to control the biosynthesis of the glycopeptide antibiotic A40926 in the actinomycete Nonomuraea sp. strain ATCC 39727. A40926 is the precursor of dalbavancin, approved for treatment of skin infections by Gram-positive bacteria. Therefore, understanding the regulation of its biosynthesis is also of industrial importance. So far, the regulatory mechanisms used to control two other similar glycopeptides (balhimycin and teicoplanin) have been elucidated, and beyond a common step, different clusters seem to have devised different strategies to control glycopeptide production. Thus, our work provides one more example of the pitfalls of deducing regulatory roles from bioinformatic analyses only, even when analyzing gene clusters directing the synthesis of structurally related compounds. T he glycopeptide antibiotic A40926 (Fig. 1A) is the precursor for dalbavancin, a semisynthetic lipoglycopeptide approved for clinical use in 2014 by the U.S. Food and Drug Administration to treat acute bacterial skin and skin structure infections caused by bacteria, such as Staphylococcus aureus (methicillin-susceptible and methicillin-resistant strains) and Streptococcus pyogenes. Compared with the structurally related glycopeptide teicoplanin, dalbavancin possesses better potency and pharmacokinetic properties.A40926 is produced by the actinomycete Nonomurae...

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A Polyketide Synthase in Glycopeptide Biosynthesis

Cited by 141 publications

References 33 publications

Insights into an Unusual Nonribosomal Peptide Synthetase Biosynthesis

Insights into an Unusual Nonribosomal Peptide Synthetase Biosynthesis

Assembling the glycopeptide antibiotic scaffold: The biosynthesis of from Streptomyces toyocaensis NRRL15009

Two Master Switch Regulators Trigger A40926 Biosynthesis in Nonomuraea sp. Strain ATCC 39727

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