A Bidomain Nonribosomal Peptide Synthetase Encoded by <i>FUM14</i> Catalyzes the Formation of Tricarballylic Esters in the Biosynthesis of Fumonisins

Zaleta-Rivera, Kathia; Xu, Chunping; Yu, Fei; Butchko, Robert A. E.; Proctor, Robert H.; Hidalgo-Lara, María Eugenia; Raza, Ashraf; Dussault, Patrick H.; Du, Liangcheng

doi:10.1021/bi052085s

Cited by 72 publications

(76 citation statements)

References 32 publications

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“…PyrG is therefore an NRPS/PKS hybrid protein. The C domain in PyrG probably forms a C-O (ester) link as has been found in fumonisin (32) and antibiotic C-1027 (33).…”

Section: Cloning Sequencing and Analysis Of The Pyr Gene Cluster-mentioning

confidence: 79%

Identification and Characterization of the Pyridomycin Biosynthetic Gene Cluster of Streptomyces pyridomyceticus NRRL B-2517

Huang

Wang

Yin

et al. 2011

Journal of Biological Chemistry

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Pyridomycin is a structurally unique antimycobacterial cyclodepsipeptide containing rare 3-(3-pyridyl)-L-alanine and 2-hydroxy-3-methylpent-2-enoic acid moieties. The biosynthetic gene cluster for pyridomycin has been cloned and identified from Streptomyces pyridomyceticus NRRL B-2517. Sequence analysis of a 42.5-kb DNA region revealed 26 putative open reading frames, including two nonribosomal peptide synthetase (NRPS) genes and a polyketide synthase gene. A special feature is the presence of a polyketide synthase-type ketoreductase domain embedded in an NRPS. Furthermore, we showed that PyrA functioned as an NRPS adenylation domain that activates 3-hydroxypicolinic acid and transfers it to a discrete peptidyl carrier protein, PyrU, which functions as a loading module that initiates pyridomycin biosynthesis in vivo and in vitro. PyrA could also activate other aromatic acids, generating three pyridomycin analogues in vivo.

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“…PyrG is therefore an NRPS/PKS hybrid protein. The C domain in PyrG probably forms a C-O (ester) link as has been found in fumonisin (32) and antibiotic C-1027 (33).…”

Section: Cloning Sequencing and Analysis Of The Pyr Gene Cluster-mentioning

confidence: 79%

Identification and Characterization of the Pyridomycin Biosynthetic Gene Cluster of Streptomyces pyridomyceticus NRRL B-2517

Huang

Wang

Yin

et al. 2011

Journal of Biological Chemistry

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“…Although the C-terminal C domain in OzmL has a catalytic motif NYFCLDG and lacks the crucial His amino acid that is believed to be indispensable for condensation of aminoacyl-ACP and peptidyl-ACP, it may still be functional, especially given that this domain is not expected to catalyze peptide bond formation but only cyclization of the Ser side chain to form a ␤-lactone ring. Some recently characterized C domains exemplified by SgcC5 (50) and Fum14 (51) have been biochemically demonstrated to catalyze ester bond formation. A similar function would be envisioned for the OzmL C-terminal C domain to be involved in ␤-lactone ring formation.…”

Section: Discussionmentioning

confidence: 99%

Oxazolomycin Biosynthesis in Streptomyces albus JA3453 Featuring an “Acyltransferase-less” Type I Polyketide Synthase That Incorporates Two Distinct Extender Units

Zhao

Coughlin

et al. 2010

Journal of Biological Chemistry

115

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The oxazolomycins (OZMs) are a growing family of antibiotics produced by several Streptomyces species that show diverse and important antibacterial, antitumor, and anti-human immunodeficiency virus activity. Oxazolomycin A is a peptidepolyketide hybrid compound containing a unique spiro-linked ␤-lactone/␥-lactam, a 5-substituted oxazole ring. The oxazolomycin biosynthetic gene cluster (ozm) was identified from Streptomyces albus JA3453 and localized to 79.5-kb DNA, consisting of 20 open reading frames that encode non-ribosomal peptide synthases, polyketide synthases (PKSs), hybrid nonribosomal peptide synthase-PKS, trans-acyltransferases (trans-ATs), enzymes for methoxymalonyl-acyl carrier protein (ACP) synthesis, putative resistance genes, and hypothetical regulation genes. In contrast to classical type I polyketide or fatty acid biosynthases, all 10 PKS modules in the gene cluster lack cognate ATs. Instead, discrete ATs OzmM (with tandem domains OzmM-AT1 and OzmM-AT2) and OzmC were equipped to carry out all of the loading functions of both malonyl-CoA and methoxymalonyl-ACP extender units. Strikingly, only OzmM-AT2 is required for OzmM activity for OZM biosynthesis, whereas OzmM-AT1 seemed to be a cryptic AT domain. The above findings, together with previous results using isotope-labeled precursor feeding assays, are assembled for the OZM biosynthesis model to be proposed. The incorporation of both malonylCoA (by OzmM-AT2) and methoxymalonyl-ACP (by OzmC) extender units seemed to be unprecedented for this class of trans-AT type I PKSs, which might be fruitfully manipulated to create structurally diverse novel compounds.

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“…SgcC5 now joins Fum14 (21) in the category of C domains that have been biochemically demonstrated to catalyze ester bond formation, but to our knowledge, SgcC5 is the first C domain to be shown to catalyze both ester and amide bond formation. SgcC5, therefore, may represent a rare event of evolution caught in action, providing an opportunity to investigate how to evolve an amide-forming C enzyme to an ester-forming enzyme or vice versa.…”

Section: Sgcc5 Is An Ester Bond-forming and Amide Bond-forming Condenmentioning

confidence: 99%

“…This includes the nonribosomal peptides kutzneride (9), valinomycin (18), and cereulide (19), and the nonribosomal peptidepolyketide hybrid cryptophycin 1 (20). Recently, the C domain of a PCP-C didomain protein Fum14 in the biosynthesis of the fungal metabolite fumonisin was shown to generate 2 tricarballylic esters by using a thioester of N-acetylcysteamine as an acyl donor mimicking the phosphopantetheinyl group of the carrier protein, and in this example, the acceptor substrate was the free alcohol of the fumonisin precursor (21). This study provided the first biochemical evidence for a C domain to catalyze ester bond formation.…”

mentioning

confidence: 99%

A free-standing condensation enzyme catalyzing ester bond formation in C-1027 biosynthesis

Lin

Lanen

Shen

2009

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

109

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Nonribosomal peptide synthetases (NRPSs) catalyze the biosynthesis of many biologically active peptides and typically are modular, with each extension module minimally consisting of a condensation, an adenylation, and a peptidyl carrier protein domain responsible for incorporation of an amino acid into the growing peptide chain. C-1027 is a chromoprotein antitumor antibiotic whose enediyne chromophore consists of an enediyne core, a deoxy aminosugar, a benzoxazolinate, and a ␤-amino acid moiety. Bioinformatics analysis suggested that the activation and incorporation of the ␤-amino acid moiety into C-1027 follows an NRPS mechanism whereby biosynthetic intermediates are tethered to the peptidyl carrier protein SgcC2. Here, we report the biochemical characterization of SgcC5, an NRPS condensation enzyme that catalyzes ester bond formation between the SgcC2-tethered (S)-3-chloro-5-hydroxy-␤-tyrosine and (R)-1-phenyl-1,2-ethanediol, a mimic of the enediyne core. SgcC5 uses (S)-3-chloro-5-hydroxy-␤-tyrosyl-SgcC2 as the donor substrate and exhibits regiospecificity for the C-2 hydroxyl group of the enediyne core mimic as the acceptor substrate. Remarkably, SgcC5 is also capable of catalyzing amide bond formation, albeit with significantly reduced efficiency, between (S)-3-chloro-5-hydroxy-␤-tyrosyl-(S)-SgcC2 and (R)-2-amino-1-phenyl-1-ethanol, an alternative enediyne core mimic bearing an amine at its C-2 position. Thus, SgcC5 is capable of catalyzing both ester and amide bond formation, providing an evolutionary link between amide-and ester-forming condensation enzymes.enediyne ͉ nonribosomal peptide synthetase N onribosomal peptide synthetases (NRPSs) are large multifunctional proteins that catalyze the synthesis of many pharmaceutically important peptides, including the anticancer drugs bleomycin and actinomycin and antibacterial antibiotics vancomycin and daptomycin. The prototypical NRPS is composed of loading, extension, and termination modules, with each extension module consisting of minimally 3 domains-an adenylation (A) domain, a peptidyl carrier protein (PCP) domain, and a condensation (C) domain (1-4). The A domain specifically selects an amino acid, activates it by formation of an aminoacyl adenylate, and transfers the activated substrate to the thiol group of the 4Ј-phosphopantetheinyl arm of the PCP domain to yield a thioester-linked amino acid. The C domain then catalyzes nucleophilic condensation between upstream and downstream PCP-tethered amino acids (also known as donor and acceptor substrates, respectively; ref. 5) to form a new amide bond, thus extending the growing peptide chain by 1 amino acid.Although NRPS catalysis is often described to follow assembly-line enzymology, wherein the amino acid sequence of the peptide product can be directly predicted from the molecular architecture of NRPS domains and modules (3, 4), recent studies have revealed numerous NRPSs that deviate from this assemblyline molecular logic (6-8). For example, variations to the standard C-A-PCP module were found in the kutzn...

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A Bidomain Nonribosomal Peptide Synthetase Encoded by FUM14 Catalyzes the Formation of Tricarballylic Esters in the Biosynthesis of Fumonisins

Cited by 72 publications

References 32 publications

Identification and Characterization of the Pyridomycin Biosynthetic Gene Cluster of Streptomyces pyridomyceticus NRRL B-2517

Identification and Characterization of the Pyridomycin Biosynthetic Gene Cluster of Streptomyces pyridomyceticus NRRL B-2517

Oxazolomycin Biosynthesis in Streptomyces albus JA3453 Featuring an “Acyltransferase-less” Type I Polyketide Synthase That Incorporates Two Distinct Extender Units

A free-standing condensation enzyme catalyzing ester bond formation in C-1027 biosynthesis

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