Evidence for a novel phosphopantetheinyl transferase domain in the polyketide synthase for enediyne biosynthesis

Murugan, Elavazhagan; Liang, Zhao‐Xun

doi:10.1016/j.febslet.2008.02.061

Cited by 28 publications

(40 citation statements)

References 29 publications

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“…1, 4b, online resource 2). These findings are agreeable with the conserved roles of PPTase in post-translational activation of fatty acid synthases, polyketide synthases, and nonribosomal peptide synthetases (Liu et al 2005;Murugan and Liang 2008;Orikasa et al 2006); suggesting that the effect of ZmsO on zeamines production is mainly through modulation of the enzymes associated with zeamines biosynthesis. Since zeamines are the main virulence factor of D. zeae to cause the pathogenicity change between wild type strain EC1 and zmsO derived strains.…”

Section: Discussionsupporting

confidence: 78%

“…Evidence is accumulating that post-translational modification of enzymes is necessary for biosynthesis of natural products by fatty acid synthases (Orikasa et al 2006), polyketide synthases (Murugan and Liang 2008), and nonribosomal peptide synthetases (Liu et al 2005). This modification is carried out by transformation of 4′-phosphopantetheine moiety from coenzyme A (CoA) to apo-carrier proteins (CPs) to form holo-carrier proteins by phosphopantetheinyl transferase (PPTase) (Beld et al 2014;Sunbul et al 2009).…”

Section: Introductionmentioning

confidence: 99%

“…Comparative analysis showed that ACPS-type PPTases usually contain about 120 amino acids carrying motif P2 [(V/I)G(V/I)D] and motif P3 [(F/W)(S/C/T/)xKE(A/S)xxK] (Lambalot et al 1996), whereas the Sfp-type PPTases contain about 220 amino acids with motifs P2 and P3 plus motif P1 [PxWPxGxxGS(M/L)THCxGY] (Sanchez et al 2001). The third type of PPTases is fused with fatty acid synthases or polyketide synthases as a functional domain (Murugan and Liang 2008;Zhang et al 2008).…”

Section: Introductionmentioning

confidence: 99%

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A Sfp-type phosphopantetheinyl transferase ZmsO is essential for zeamines production and the virulence of Dickeya zeae

et al. 2016

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Zeamines are family of potent antibiotics and virulence determinants produced by the rice foot rot bacterial pathogen Dickeya zeae. So zeamines are important for the pathogenesis of D. zeae and development of new strategies against this devastating disease. In this study, we show that production of zeamines is positively modulated by ZmsO, which is a conserved Sfp-type phosphopantetheinyl thransferase (PPTase) associated with post-translational activation of fatty acid synthases and polyketide synthases. Deletion of zmsO significantly abolished zeamines production and attenuated the virulence of D. zeae without affect the growth rate. The zmsO gene is located at the upstream of zmsA and zmsK in the same gene cluster. Consistent with its role in post-translational modification, deletion of zmsO did not affect the transcriptional expression of zmsA and zmsK. In trans expression of the pcpS gene from Pseudomonas aeruginosa, which encodes a Sfp-type PPTase, in the zmsO deletion mutant could also fully restore the zeamines production and bacterial virulence, establishing the important role of Sfp-type PPTase in the zeamines biosynthesis and pathogenicity of D. zeae.

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Section: Discussionsupporting

confidence: 78%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Sfp-type phosphopantetheinyl transferase ZmsO is essential for zeamines production and the virulence of Dickeya zeae

et al. 2016

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“…The spore preparation of S. chattanoogensis L10 on YMG agar (0.4% yeast extract, 1% malt extract, 0.4% glucose, 2% agar, 0.2% CaCO 3 [pH 7.2]), the examination of NTM production in YEME with 4% glucose (0.3% yeast extract, 0.3% malt extract, 0.5% tryptone, 4% glucose), and DNA manipulations in S. chattanoogensis L10 were carried out according to standard procedures (25). Escherichia coli-Streptomyces conjugation, fermentation of S. chattanoogensis L10 and its recombinant strains, and quantification of NTM production were performed as described previously (11,15,20).…”

Section: Methodsmentioning

confidence: 99%

Improvement of Natamycin Production by Engineering of Phosphopantetheinyl Transferases in Streptomyces chattanoogensis L10

Jiang

Wang

Ran

et al. 2013

Appl Environ Microbiol

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Phosphopantetheinyl transferases (PPTases) are essential to the activities of type I/II polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs) through converting acyl carrier proteins (ACPs) in PKSs and peptidyl carrier proteins (PCPs) in NRPSs from inactive apo-forms into active holo-forms, leading to biosynthesis of polyketides and nonribosomal peptides. The industrial natamycin (NTM) producer,Streptomyces chattanoogensisL10, contains two PPTases (SchPPT and SchACPS) and five PKSs. Biochemical characterization of these two PPTases shows that SchPPT catalyzes the phosphopantetheinylation of ACPs in both type I PKSs and type II PKSs, SchACPS catalyzes the phosphopantetheinylation of ACPs in type II PKSs and fatty acid synthases (FASs), and the specificity of SchPPT is possibly controlled by its C terminus. Inactivation of SchPPT inS. chattanoogensisL10 abolished production of NTM but not the spore pigment, while overexpression of the SchPPT gene not only increased NTM production by about 40% but also accelerated productions of both NTM and the spore pigment. Thus, we elucidated a comprehensive phosphopantetheinylation network of PKSs and improved polyketide production by engineering the cognate PPTase in bacteria.

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“…The release of the product catalyzed by the putative thioesterase SgcE10 presumably occurs through a combination of hydrolysis, decarboxylation, and dehydration steps. Recent biochemical studies of the iterative PKS (CalE8) from the biosynthetic pathway of calicheamicin also provided insight into the early steps of 10-membered enediyne biosynthesis (13,14). It was observed that CalE8 produced a linear carbonyl-conjugated polyene (3,5,7,9,11,13-pentadecen-2-one (1)) with the assistance of the putative thioesterase CalE7 (Fig.…”

mentioning

confidence: 99%

Structure and Catalytic Mechanism of the Thioesterase CalE7 in Enediyne Biosynthesis

Kotaka¹,

Kong²,

Qureshi

et al. 2009

Journal of Biological Chemistry

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The biosynthesis of the enediyne moiety of the antitumor natural product calicheamicin involves an iterative polyketide synthase (CalE8) and other ancillary enzymes. In the proposed mechanism for the early stage of 10-membered enediyne biosynthesis, CalE8 produces a carbonyl-conjugated polyene with the assistance of a putative thioesterase (CalE7). We have determined the x-ray crystal structure of CalE7 and found that the subunit adopts a hotdog fold with an elongated and kinked substrate-binding channel embedded between two subunits. The 1.75-Å crystal structure revealed that CalE7 does not contain a critical catalytic residue (Glu or Asp) conserved in other hotdog fold thioesterases. Based on biochemical and site-directed mutagenesis studies, we proposed a catalytic mechanism in which the conserved Arg 37 plays a crucial role in the hydrolysis of the thioester bond, and that Tyr 29 and a hydrogen-bonded water network assist the decarboxylation of the ␤-ketocarboxylic acid intermediate. Moreover, computational docking suggested that the substrate-binding channel binds a polyene substrate that contains a single cis double bond at the C4/C5 position, raising the possibility that the C4‫؍‬C5 double bond in the enediyne moiety could be generated by the iterative polyketide synthase. Together, the results revealed a hotdog fold thioesterase distinct from the common type I and type II thioesterases associated with polyketide biosynthesis and provided interesting insight into the enediyne biosynthetic mechanism.Enediyne natural products represent a family of structurally unique secondary metabolites with potent antitumor and antibiotic activities. Based on the structure of the bicyclic enediyne core, enediyne natural products are categorized into two groups with either a 9-or 10-membered enediyne moiety (1, 2). The antitumor activity of enediyne natural products derives from their capacity to induce chromosomal DNA cleavage through an oxidative radical mechanism (3). The biosynthetic mechanism for the enediyne moiety has been, however, elusive despite clues gleaned from early isotope-feeding experiments (4, 5). Pioneering genetic studies of the biosynthesis of calicheamicin and C-1027 from two research groups yielded major insights into the biosynthetic pathways, suggesting that an iterative polyketide synthase (PKS) 5 plays a central role in the assembly of both the 9-and 10-membered enediyne moieties (6, 7). The gene clusters also contain open reading frames encoding hypothetical proteins for the downstream processing of the PKS product. The involvement of similar genes in enediyne biosynthesis was later confirmed for neocarzinostatin, maduropeptin, dynemicin, and several putative enediyne natural products in soil and marine microorganisms (8 -11). Recently, based on the study on the 9-membered enediynecontaining C-1027, Shen and coworkers found that the iterative PKS (SgcE) and the putative thioesterase (SgcE10) generated a conjugated polyene (1,3,5,7,9,11,13-pentadecaheptaene) through an ACP-tethered 3-hydroxy-4...

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Evidence for a novel phosphopantetheinyl transferase domain in the polyketide synthase for enediyne biosynthesis

Cited by 28 publications

References 29 publications

A Sfp-type phosphopantetheinyl transferase ZmsO is essential for zeamines production and the virulence of Dickeya zeae

A Sfp-type phosphopantetheinyl transferase ZmsO is essential for zeamines production and the virulence of Dickeya zeae

Improvement of Natamycin Production by Engineering of Phosphopantetheinyl Transferases in Streptomyces chattanoogensis L10

Structure and Catalytic Mechanism of the Thioesterase CalE7 in Enediyne Biosynthesis

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