scholarly journals Directed biosynthesis of fluorinated polyketides

2021 ◽  
Author(s):  
Alexander Rittner ◽  
Mirko Joppe ◽  
Jennifer J. Schmidt ◽  
Lara Maria Mayer ◽  
Elia Heid ◽  
...  
Keyword(s):  
Fatty Acid Synthase ◽  
Polyketide Synthase ◽  
Coenzyme A ◽  
Chain Extension ◽  
Type I ◽  
Complex Molecules ◽  
Site Specific ◽  
Directed Biosynthesis ◽  
Polyketide Chain ◽  
Selection Of

Modification of polyketides with fluorine offers a promising approach to develop new pharmaceuticals. While synthetic chemical methods for site-specific incorporation of fluorine in complex molecules have improved in recent years, approaches for the direct biosynthetic fluorination of natural compounds are still rare. Herein, we present a broadly applicable approach for site-specific, biocatalytic derivatization of polyketides with fluorine. Specifically, we exchanged the native acyltransferase domain (AT) of a polyketide synthase (PKS), which acts as the gatekeeper for selection of extender units, with an evolutionarily related but substrate tolerant domain from metazoan type I fatty acid synthase (FAS). The resulting PKS/FAS hybrid can utilize fluoromalonyl coenzyme A and fluoromethylmalonyl coenzyme A for polyketide chain extension, introducing fluorine or fluoro-methyl disubstitutions in polyketide scaffolds. Addition of a fluorine atom is often a decisive factor toward developing superior properties in next-generation antibiotics, including the macrolide solithromycin. We demonstrate the feasibility of our approach in the semisynthesis of a fluorinated derivative of the macrolide antibiotic YC-17.

scholarly journals Cloning, Sequencing, and Functional Analysis of an Iterative Type I Polyketide Synthase Gene Cluster for Biosynthesis of the Antitumor Chlorinated Polyenone Neocarzilin in “Streptomyces carzinostaticus”

2004 ◽  
Vol 48 (9) ◽  
pp. 3468-3476 ◽  
Author(s):  
Miyuki Otsuka ◽  
Koji Ichinose ◽  
Isao Fujii ◽  
Yutaka Ebizuka
Keyword(s):  
Gene Cluster ◽  
Polyketide Synthase ◽  
Streptomyces Coelicolor ◽  
Open Reading Frames ◽  
Modular Organization ◽  
Type I ◽  
Orf3 Protein ◽  
Polyketide Synthase Gene ◽  
Polyketide Chain ◽  
Reading Frames

ABSTRACT Neocarzilins (NCZs) are antitumor chlorinated polyenones produced by “Streptomyces carzinostaticus” var. F-41. The gene cluster responsible for the biosynthesis of NCZs was cloned and characterized. DNA sequence analysis of a 33-kb region revealed a cluster of 14 open reading frames (ORFs), three of which (ORF4, ORF5, and ORF6) encode type I polyketide synthase (PKS), which consists of four modules. Unusual features of the modular organization is the lack of an obvious acyltransferase domain on modules 2 and 4 and the presence of longer interdomain regions more than 200 amino acids in length on each module. Involvement of the PKS genes in NCZ biosynthesis was demonstrated by heterologous expression of the cluster in Streptomyces coelicolor CH999, which produced the apparent NCZ biosynthetic intermediates dechloroneocarzillin A and dechloroneocarzilin B. Disruption of ORF5 resulted in a failure of NCZ production, providing further evidence that the cluster is essential for NCZ biosynthesis. Mechanistic consideration of NCZ formation indicates the iterative use of at least one module of the PKS, which subsequently releases its product by decarboxylation to generate an NCZ skeleton, possibly catalyzed by a type II thioesterase encoded by ORF7. This is a novel type I PKS system of bacterial origin for the biosynthesis of a reduced polyketide chain. Additionally, the protein encoded by ORF3, located upstream of the PKS genes, closely resembles the FADH2-dependent halogenases involved in the formation of halometabolites. The ORF3 protein could be responsible for the halogenation of NCZs, presenting a unique example of a halogenase involved in the biosynthesis of an aliphatic halometabolite.

scholarly journals Engineering of a minimal modular polyketide synthase, and targeted alteration of the stereospecificity of polyketide chain extension

1998 ◽  
Vol 5 (8) ◽  
pp. 407-412 ◽  
Author(s):  
Ines Böhm ◽  
Inès E. Holzbaur ◽  
Ulf Hanefeld ◽  
Jesus Cortési ◽  
Jim Staunton ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Chain Extension ◽  
Polyketide Chain ◽  
Modular Polyketide Synthase

scholarly journals Biosynthesis of Cell Envelope-Associated Phenolic Glycolipids in Mycobacterium marinum

2015 ◽  
Vol 197 (6) ◽  
pp. 1040-1050 ◽  
Author(s):  
Olivia Vergnolle ◽  
Sivagami Sundaram Chavadi ◽  
Uthamaphani R. Edupuganti ◽  
Poornima Mohandas ◽  
Catherine Chan ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Cell Envelope ◽  
Acyl Chain ◽  
Deep Understanding ◽  
Fatty Acyl ◽  
Chain Extension ◽  
Type I ◽  
Content Type ◽  
Iterative Polyketide Synthase ◽  
Phenolic Glycolipids

Phenolic glycolipids (PGLs) are polyketide synthase-derived glycolipids unique to pathogenic mycobacteria. PGLs are found in several clinically relevant species, including variousMycobacterium tuberculosisstrains,Mycobacterium leprae, and several nontuberculous mycobacterial pathogens, such asM. marinum. Multiple lines of investigation implicate PGLs in virulence, thus underscoring the relevance of a deep understanding of PGL biosynthesis. We report mutational and biochemical studies that interrogate the mechanism by which PGL biosynthetic intermediates (p-hydroxyphenylalkanoates) synthesized by the iterative polyketide synthase Pks15/1 are transferred to the noniterative polyketide synthase PpsA for acyl chain extension inM. marinum. Our findings support a model in which the transfer of the intermediates is dependent on ap-hydroxyphenylalkanoyl-AMP ligase (FadD29) acting as an intermediary between the iterative and the noniterative synthase systems. Our results also establish thep-hydroxyphenylalkanoate extension ability of PpsA, the first-acting enzyme of a multisubunit noniterative polyketide synthase system. Notably, this noniterative system is also loaded with fatty acids by a specific fatty acyl-AMP ligase (FadD26) for biosynthesis of phthiocerol dimycocerosates (PDIMs), which are nonglycosylated lipids structurally related to PGLs. To our knowledge, the partially overlapping PGL and PDIM biosynthetic pathways provide the first example of two distinct, pathway-dedicated acyl-AMP ligases loading the same type I polyketide synthase system with two alternate starter units to produce two structurally different families of metabolites. The studies reported here advance our understanding of the biosynthesis of an important group of mycobacterial glycolipids.

scholarly journals Cooperation between a Coenzyme A-Independent Stand-Alone Initiation Module and an Iterative Type I Polyketide Synthase during Synthesis of Mycobacterial Phenolic Glycolipids

2009 ◽  
Vol 131 (46) ◽  
pp. 16744-16750 ◽  
Author(s):  
Weiguo He ◽  
Clifford E. Soll ◽  
Sivagami Sundaram Chavadi ◽  
Guangtao Zhang ◽  
J. David Warren ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Coenzyme A ◽  
Type I ◽  
Phenolic Glycolipids

scholarly journals Quantification ofN-acetylcysteamine activated methylmalonate incorporation into polyketide biosynthesis

2013 ◽  
Vol 9 ◽  
pp. 664-674 ◽  
Author(s):  
Stephan Klopries ◽  
Uschi Sundermann ◽  
Frank Schulz
Keyword(s):  
Malonic Acid ◽  
Polyketide Synthase ◽  
Coenzyme A ◽  
Building Blocks ◽  
Substrate Analogues ◽  
Experimental Alteration ◽  
Malonic Acid Derivatives ◽  
Acid Thioesters ◽  
Selection Of

Polyketides are biosynthesized through consecutive decarboxylative Claisen condensations between a carboxylic acid and differently substituted malonic acid thioesters, both tethered to the giant polyketide synthase enzymes. Individual malonic acid derivatives are typically required to be activated as coenzyme A-thioesters prior to their enzyme-catalyzed transfer onto the polyketide synthase. Control over the selection of malonic acid building blocks promises great potential for the experimental alteration of polyketide structure and bioactivity. One requirement for this endeavor is the supplementation of the bacterial polyketide fermentation system with tailored synthetic thioester-activated malonates. The membrane permeableN-acetylcysteamine has been proposed as a coenzyme A-mimic for this purpose. Here, the incorporation efficiency into different polyketides ofN-acetylcysteamine activated methylmalonate is studied and quantified, showing a surprisingly high and transferable activity of these polyketide synthase substrate analogues in vivo.

Enzymatic Synthesis of 1,3,6,8-Tetrahydroxynaphthalene Solely from Malonyl Coenzyme A by a Fungal Iterative Type I Polyketide Synthase PKS1†

Biochemistry ◽  
2000 ◽  
Vol 39 (30) ◽  
pp. 8853-8858 ◽  
Author(s):  
Isao Fujii ◽  
Yuichiro Mori ◽  
Akira Watanabe ◽  
Yasuyuki Kubo ◽  
Gento Tsuji ◽  
...  
Keyword(s):  
Enzymatic Synthesis ◽  
Polyketide Synthase ◽  
Coenzyme A ◽  
Type I

scholarly journals Evidence for an iterative module in chain elongation on the azalomycin polyketide synthase

2016 ◽  
Vol 12 ◽  
pp. 2164-2172 ◽  
Author(s):  
Hui Hong ◽  
Yuhui Sun ◽  
Yongjun Zhou ◽  
Emily Stephens ◽  
Markiyan Samborskyy ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Polyketide Synthase ◽  
Strong Support ◽  
Artificial Chromosome ◽  
Chain Extension ◽  
Chain Elongation ◽  
Starter Unit ◽  
Polyketide Chain ◽  
Streptomyces Olivaceus ◽  
Streptomyces Malaysiensis

The assembly-line synthases that produce bacterial polyketide natural products follow a modular paradigm in which each round of chain extension is catalysed by a different set or module of enzymes. Examples of deviation from this paradigm, in which a module catalyses either multiple extensions or none are of interest from both a mechanistic and an evolutionary viewpoint. We present evidence that in the biosynthesis of the 36-membered macrocyclic aminopolyol lactones (marginolactones) azalomycin and kanchanamycin, isolated respectively from Streptomyces malaysiensis DSM4137 and Streptomyces olivaceus Tü4018, the first extension module catalyses both the first and second cycles of polyketide chain extension. To confirm the integrity of the azl gene cluster, it was cloned intact on a bacterial artificial chromosome and transplanted into the heterologous host strain Streptomyces lividans, which does not possess the genes for marginolactone production. When furnished with 4-guanidinobutyramide, a specific precursor of the azalomycin starter unit, the recombinant S. lividans produced azalomycin, showing that the polyketide synthase genes in the sequenced cluster are sufficient to accomplish formation of the full-length polyketide chain. This provides strong support for module iteration in the azalomycin and kanchanamycin biosynthetic pathways. In contrast, re-sequencing of the gene cluster for biosynthesis of the polyketide β-lactone ebelactone in Streptomyces aburaviensis has shown that, contrary to a recently-published proposal, the ebelactone polyketide synthase faithfully follows the colinear modular paradigm.

scholarly journals Reconstitution of the FK228 Biosynthetic Pathway Reveals Cross Talk between Modular Polyketide Synthases and Fatty Acid Synthase

2010 ◽  
Vol 77 (4) ◽  
pp. 1501-1507 ◽  
Author(s):  
Shane R. Wesener ◽  
Vishwakanth Y. Potharla ◽  
Yi-Qiang Cheng
Keyword(s):  
Fatty Acid ◽  
Cross Talk ◽  
Biosynthetic Pathway ◽  
Fatty Acid Synthase ◽  
Polyketide Synthase ◽  
Fatty Acid Biosynthesis ◽  
Chain Elongation ◽  
E Coli ◽  
Genetic Components ◽  
Polyketide Chain

ABSTRACTFunctional cross talk between fatty acid biosynthesis and secondary metabolism has been discovered in several cases in microorganisms; none of them, however, involves a modular biosynthetic enzyme. Previously, we reported a hybrid modular nonribosomal peptide synthetase (NRPS)-polyketide synthase (PKS) pathway for the biosynthesis of FK228 anticancer depsipeptide inChromobacterium violaceumstrain 968. This pathway contains two PKS modules on the DepBC enzymes that lack a functional acyltransferase (AT) domain, and no apparent AT-encoding gene exists within the gene cluster or its vicinity. We report here that, through reconstitution of the FK228 biosynthetic pathway inEscherichia colicells, two essential genes,fabD1andfabD2, both encoding a putative malonyl coenzyme A (CoA) acyltransferase component of the fatty acid synthase complex, are positively identified to be involved in FK228 biosynthesis. Either gene product appears sufficient to complement the AT-less PKS modules on DepBC for polyketide chain elongation. Concurrently, a gene (sfp) encoding a putative Sfp-type phosphopantetheinyltransferase was identified to be necessary for FK228 biosynthesis as well. Most interestingly, engineeredE. colistrains carrying variable genetic components produced significant levels of FK228 under both aerobic and anaerobic cultivation conditions. Discovery of thetranscomplementation of modular PKSs by housekeeping ATs reveals natural product biosynthesis diversity. Moreover, demonstration of anaerobic production of FK228 by an engineered facultative bacterial strain validates our effort toward the engineering of novel tumor-targeting bioagents.

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