Polyketide Synthase Modules Redefined

Keatinge‐Clay, Adrian T.

doi:10.1002/anie.201701281

Cited by 73 publications

(67 citation statements)

References 28 publications

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“…These interactions enable specific transformations to occur on the acyl chain covalently shuttled by that ACP. This holds for both Type II systems in which each domain is encoded on a separate polypeptide, as in most bacterial fatty acid synthases (FASs), as well as Type I systems in which several domains are encoded on the same polypeptide, as in biosynthetic assembly lines formed by polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) machinery …”

Section: Introductionmentioning

confidence: 99%

“…The biosynthetic community is working to decipher the logic of 2 large classes of assembly lines that contain PKS machinery— cis ‐AT assembly lines that primarily rely on embedded ATs and trans ‐AT assembly lines that primarily rely on separately encoded ATs . From the sequencing of the erythromycin synthase in 1990 until last year, the boundaries of the modules of cis ‐AT assembly lines were incorrectly defined—comparisons with the domain organization of the mammalian FAS had led to the module being defined with KS at its upstream boundary and ACP at its downstream boundary.…”

Section: Introductionmentioning

confidence: 99%

“…From the sequencing of the erythromycin synthase in 1990 until last year, the boundaries of the modules of cis ‐AT assembly lines were incorrectly defined—comparisons with the domain organization of the mammalian FAS had led to the module being defined with KS at its upstream boundary and ACP at its downstream boundary. The processing enzymes in several cis ‐AT assembly lines have been shown to evolutionarily co‐migrate with the KS downstream of them, leading to the redefinition of cis ‐AT modules as possessing a downstream KS . When trans ‐AT assembly lines started to be discovered 15 years ago, often from difficult‐to‐culture, symbiotic bacteria, the use of cis ‐AT nomenclature resulted in their modules being incorrectly defined as well .…”

Section: Introductionmentioning

confidence: 99%

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The modules of trans‐acyltransferase assembly lines redefined with a central acyl carrier protein

Wood

Keatinge‐Clay

2018

Proteins

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Here, the term "module" is redefined for trans-acyltransferase (trans-AT) assembly lines to agree with how its domains cooperate and evolutionarily co-migrate. The key domain in both the polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) modules of assembly lines is the acyl carrier protein (ACP). ACPs not only relay growing acyl chains through the assembly line but also collaborate with enzymes in modules, both in cis and in trans, to add a specific chemical moiety. A ketosynthase (KS) downstream of ACP often plays the role of gatekeeper, ensuring that only a single intermediate generated by the enzymes of a module is passed downstream. Bioinformatic analysis of 526 ACPs from 33 characterized trans-AT assembly lines reveals ACPs from the same module type generally clade together, reflective of the co-evolution of these domains with their cognate enzymes. While KSs downstream of ACPs from the same module type generally also clade together, KSs upstream of ACPs do not-in disagreement with the traditional definition of a module. Beyond nomenclature, the presented analysis impacts our understanding of module function, the evolution of assembly lines, pathway prediction, and assembly line engineering.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The modules of trans‐acyltransferase assembly lines redefined with a central acyl carrier protein

Wood

Keatinge‐Clay

2018

Proteins

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“…21 Oxyanion formation in the EI reaction suggests oxyanion formation in the PS reaction. An alternative hypothesis is that a four-membered transition state is formed between the OH of the ζ-hydroxyl group and C α /C β , as proposed for eukaryotic hydratase 2 from the crotonase superfamily; 30 however, as several other crotonase enzymes, such as Δ 3,5 ,Δ 2,4 -dienoyl-CoA isomerase, 4-chlorobenzyl-CoA dehalogenase, and methylmalonyl-CoA decarboxylase, apparently conduct catalysis through an oxyanion intermediate, eukaryotic hydratase 2 may also proceed through this route. Without any experimental support for a concerted mechanism in related systems, PS catalysis more likely proceeds through an oxyanion intermediate.…”

Section: Resultsmentioning

confidence: 99%

“…5,16−18 The SorPS9 active site contains a histidine and an asparagine at positions equivalent to those of the catalytic histidine and active site aspartate of DHs. Functional assays of SorPS9 point mutants with trans - α/β -unsaturated, ζ-hydroxyacyl NAC thioester substrates reveal the critical activity of the histidine and the ancillary role of the neighboring asparagine.…”

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confidence: 99%

Structural and Functional Studies of a Pyran Synthase Domain from a trans-Acyltransferase Assembly Line

et al. 2018

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trans-Acyltransferase assembly lines possess enzymatic domains often not observed in their better characterized cisacyltransferase counterparts. Within this repertoire of largely unexplored biosynthetic machinery is a class of enzymes called the pyran synthases that catalyze the formation of five- and sixmembered cyclic ethers from diverse polyketide chains. The 1.55 Å resolution crystal structure of a pyran synthase domain excised from the ninth module of the sorangicin assembly line highlights the similarity of this enzyme to the ubiquitous dehydratase domain and provides insight into the mechanism of ring formation. Functional assays of point mutants reveal the central importance of the active site histidine that is shared with the dehydratases as well as the supporting role of a neighboring semiconserved asparagine.

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Reshaping the 2‐Pyrone Synthase Active Site for Chemoselective Biosynthesis of Polyketides

Mirts

Yook

et al. 2022

Angewandte Chemie

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Engineering enzymes with novel reactivity and applying them in metabolic pathways to produce valuable products are quite challenging due to the intrinsic complexity of metabolic networks and the need for high in vivo catalytic efficiency. Triacetic acid lactone (TAL), naturally generated by 2‐pyrone synthase (2PS), is a platform molecule that can be produced via microbial fermentation and further converted into value‐added products. However, these conversions require extra synthetic steps under harsh conditions. We herein report a biocatalytic system for direct generation of TAL derivatives under mild conditions with controlled chemoselectivity by rationally engineering the 2PS active site and then rewiring the biocatalytic pathway in the metabolic network of E. coli to produce high‐value products, such as kavalactone precursors, with yields up to 17 mg/L culture. Computer modeling indicates sterics and hydrogen‐bond interactions play key roles in tuning the selectivity, efficiency and yield.

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Polyketide Synthase Modules Redefined

Cited by 73 publications

References 28 publications

The modules of trans‐acyltransferase assembly lines redefined with a central acyl carrier protein

The modules of trans‐acyltransferase assembly lines redefined with a central acyl carrier protein

Structural and Functional Studies of a Pyran Synthase Domain from a trans-Acyltransferase Assembly Line

Reshaping the 2‐Pyrone Synthase Active Site for Chemoselective Biosynthesis of Polyketides

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