Molecular Analysis of the Benastatin Biosynthetic Pathway and Genetic Engineering of Altered Fatty Acid−Polyketide Hybrids

Xu, Zhongli; Schenk, and Angéla; Hertweck, Christian

doi:10.1021/ja069045b

Cited by 71 publications

(69 citation statements)

References 78 publications

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“…Homologe dieser Enzyme sind charakteristisch für die meisten aromatischen PolyketidBiosynthesewege, die durch Nichtacetyl-Startereinheiten eingeleitet werden, wie jene, die zu Anthracyclinen, R1128 und Frenolicin führen. [54] Neuere Beispiele für die Verwendung von kurzen Fettsäuren als Starter sind die Hedamycin- (13), [63] Fredericamycin-, [64] Benastatin- [65] und AlnumycinBiosynthesewege. [66] Zusätzliche ACP-und AT-Komponenten können beteiligt sein, um die Acyl-Einheiten anzubinden oder zu übertragen, werden aber nicht in allen Fällen gefunden.…”

Section: Polyphenol-diversifizierung Durch Nichtacetylstartereinheitenunclassified

Die biosynthetische Grundlage der Polyketid‐Vielfalt

Hertweck

2009

Angewandte Chemie

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Molekulares Lego: Polyketide bilden eine äußerst vielfältige Gruppe von Naturstoffen mit faszinierenden Kohlenstoffgerüsten (siehe Beispiele), die aus einfachen Acyl‐Bausteinen synthetisiert werden. Das Zusammenspiel von Chemie, Biochemie und Genetik lieferte wichtige Einblicke in die Programmierung des Polyketid‐Aufbaus und die beteiligten, komplexen Enzymsysteme. Dieser Aufsatz behandelt aktuelle Entwicklungen auf diesem Forschungsgebiet.magnified imagePolyketide bilden eine der größten Naturstoffklassen. Viele dieser Verbindungen – oder Derivate davon – sind wichtige Therapeutika, aber viele sind auch berüchtigte, Lebensmittel verderbende Toxine oder Virulenzfaktoren. Besonders bemerkenswert ist, dass sich die Verbindungen dieser heterogenen Gruppe, zu denen Polyether, Polyene, Polyphenole, Makrolide und Endiine gehören, hauptsächlich von einem der einfachsten natürlichen Bausteine ableiten: Essigsäure. Chemische, genetische und biochemische Untersuchungen haben Aufschluss über die Biosyntheseprogramme gegeben, die zur enormen Strukturvielfalt von Polyketiden führen. Dieser Aufsatz behandelt kürzlich aufgeklärte Biosynthesemechanismen, nach denen höchst unterschiedliche und komplexe Moleküle synthetisiert werden.

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Section: Polyphenol-diversifizierung Durch Nichtacetylstartereinheitenunclassified

Die biosynthetische Grundlage der Polyketid‐Vielfalt

Hertweck

2009

Angewandte Chemie

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“…However, all of them to date were annotated as proteins of unknown functions. These include Prm-Orf5 (29.5% identity to FdmM, 42.3% identity to FdmM1) in the pradimicin A (2) biosynthetic pathway (7), LlpB (30.1% identity to FdmM, 50% identity to FdmM1), and LlpQ (41.2% identity to FdmM, 36.1% identity to FdmM1) in the lysolipin X (3) biosynthetic pathway (8), GrhM (42.2% identity to FdmM, 31% identity to FdmM1) in the griseorhodin A (4) biosynthetic pathway (9), BenG (28.6% identity to FdmM, 26.5% identity to FdmM1) in the benastatin A (5) biosynthetic pathway (10), and RubQ (43.8% identity to FdmM, 31.6% identity to FdmM1) in the ␥-rubromycin (6) biosynthetic pathway (supplemental Fig. S1 and Ref.…”

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In Vivo Investigation of the Roles of FdmM and FdmM1 in Fredericamycin Biosynthesis Unveiling a New Family of Oxygenases

Chen

Wendt-Pienkoski

Rajski

et al. 2009

Journal of Biological Chemistry

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Fredericamycin (FDM) A, a highly oxidized aromatic pentadecaketide natural product, exhibits potent cytotoxicity and has been studied as a new anticancer drug lead. The FDM biosynthetic gene cluster has been previously cloned from Streptomyces griseus ATCC 49344 and successfully expressed in the heterologous host Streptomyces albus J1074. The fdmM and fdmM1 genes code for two proteins with high sequence homology to each other but unknown function. In-frame deletion of each of the genes from the fdm cluster was accomplished in the S. albus host. Each mutant failed to produce FDM A and the key biosynthetic intermediate FDM E but produced various new metabolites, the titers of which were dramatically increased via overexpression of an fdm pathway-specific activator fdmR1. The ⌬fdmM mutant strain accumulated three new compounds FDM M-1, FDM M-2, and FDM M-3, whereas the ⌬fdmM1 mutant strain produced one new compound FDM M1-1. Isolation and structural characterization of these compounds enable us to propose that FdmM and FdmM1 catalyze the C-6 and C-8 hydroxylations for FDM biosynthesis, respectively. Homologs of FdmM and FdmM1 can be found in biosynthetic gene clusters of many other aromatic polyketides, ranging from dodecaketides to pentadecaketides, but to date all of them were annotated as proteins of unknown function. Based on the findings reported here for FdmM and FdmM1, we now propose similar functions for those proteins, and FdmM and FdmM1 therefore represent an emerging family of novel oxygenases responsible for hydroxylation of aromatic polyketide natural products.Studies on natural product biosynthetic pathways have uncovered a number of intriguing oxygenases. For instance, the first reported metal-and cofactor-free monooxygenase TcmH is responsible for quinone moiety formation in tetracenomycin D3 and represents a rapidly growing family of antibiotic biosynthetic monooxygenases (1, 2). Another example is DpgC, which is involved in the biosynthesis of vancomycin building block 3, 5-dihydroxyphenylglycine. The crystal structure of DpgC has been used to understand the reaction mechanism of metal-and cofactor-free dioxygenases (3). MomA, involved in mompain biosynthesis (4) is a metal ion-dependent monooxygenase belonging to the cupin superfamily of oxidases bearing no prosthetic group. The discovery of enzymes such as TcmH, DpgC, and MomA continually encourage the community to search for and characterize new oxygenases from proteins of unknown functions involved in natural products biosynthesis.Whereas octaketides (C-16), nonaketides (C-18), and decaketides (C-20) are the most common aromatic polyketide natural products whose biosyntheses have been extensively investigated, aromatic polyketides ranging from dodecaketides (C-24) to pentadecaketides (C-30) are also known including biologically active compounds such as fredericamycin (FDM) 2 A (1) (5, 6), pradimicin A (2) (7), lysolipin X (3) (8), griseorhodin A (4) (9), benastatin A (5) (10), and ␥-rubromycin (6) ( Fig. 1 and Ref. 11). The highly oxidized nat...

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“…The C-methylation reaction is likely catalyzed by one of two methyltransferases in the oxy gene cluster (OxyF and OxyT), both containing putative S-adenosylmethionine binding domains. C-Methyltransferases are found infrequently among bacterial type II PKSs, including MtmMII in the mithramycin gene cluster (24), RemG and RemH from the resistomycin gene cluster (25), and the recently identified BenF and NapB2 from the benastatin (26) and napyradiomycin (27) gene clusters, respectively. All these C-methyltransferases modify the respective aromatic aglycons with unique regiospecificity.…”

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Investigation of Early Tailoring Reactions in the Oxytetracycline Biosynthetic Pathway

Zhang

Watanabe

Wang

et al. 2007

Journal of Biological Chemistry

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Tetracyclines are aromatic polyketides biosynthesized by bacterial type II polyketide synthases. The amidated tetracycline backbone is biosynthesized by the minimal polyketide synthases and an amidotransferase homologue OxyD. Biosynthesis of the key intermediate 6-methylpretetramid requires two early tailoring steps, which are cyclization of the linearly fused tetracyclic scaffold and regioselective C-methylation of the aglycon. Using a heterologous host (CH999)/vector pair, we identified the minimum set of enzymes from the oxytetracycline biosynthetic pathway that is required to afford 6-methylpretetramid in vivo. Only two cyclases (OxyK and OxyN) are necessary to completely cyclize and aromatize the amidated tetracyclic aglycon. Formation of the last ring via C-1/C-18 aldol condensation does not require a dedicated fourth-ring cyclase, in contrast to the biosynthetic mechanism of other tetracyclic aromatic polyketides, such as daunorubicin and tetracenomycin. Acetylderived polyketides do not undergo spontaneous fourth-ring cyclization and form only anthracene carboxylic acids as demonstrated both in vivo and in vitro. OxyF was identified to be the C-6 C-methyltransferase that regioselectively methylates pretetramid to yield 6-methylpretetramid. Reconstitution of 6-methylpretetramid in a heterologous host sets the stage for a more systematic investigation of additional tetracycline downstream tailoring enzymes and is a key step toward the engineered biosynthesis of tetracycline analogs.Tetracyclines are aromatic polyketides biosynthesized by soil-borne Streptomyces bacteria (1, 2). It is well known that the carbon skeleton of an aromatic polyketide is assembled through stepwise decarboxylative condensation of malonate equivalents catalyzed by the minimal PKS, 2 which consists of a ketosynthase/chain length factor heterodimer (KS-CLF or KS ␣ -KS ␤ ), an acyl-carrier protein (ACP), and a malonyl-CoA:ACP acyltransferase (3). Dedicated tailoring enzymes transform the highly reactive poly-␤-ketone backbone into fused, richly substituted compounds (4). The biosynthesis of tetracyclines has been studied using blocked mutants of the chlorotetracycline producer Streptomyces aureofaciens (5-11). However, the underlying enzymology of several key tailoring steps that give rise to its structural features remain unresolved, including cyclization of the tetracyclic scaffold and C-methylation of C-6 to afford the key intermediate 6-methylpretetramid (Fig. 1). The oxytetracycline (oxy) biosynthetic gene cluster from Streptomyces rimosus has been completely sequenced, hence allowing a thorough investigation of the biochemical basis of these features (12-16).During tetracycline biosynthesis, the C-9 reduced decaketide backbone first undergoes C-7/C-12 intramolecular aldol condensation to fix the regioselectivity of the D ring followed by three sequential cyclization reactions to form a fully aromatized, tetracyclic intermediate pretetramid (Fig. 1). Biosynthesis of aureolic acids such as mithramycin presumably follows identical c...

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Molecular Analysis of the Benastatin Biosynthetic Pathway and Genetic Engineering of Altered Fatty Acid−Polyketide Hybrids

Cited by 71 publications

References 78 publications

Die biosynthetische Grundlage der Polyketid‐Vielfalt

Die biosynthetische Grundlage der Polyketid‐Vielfalt

In Vivo Investigation of the Roles of FdmM and FdmM1 in Fredericamycin Biosynthesis Unveiling a New Family of Oxygenases

Investigation of Early Tailoring Reactions in the Oxytetracycline Biosynthetic Pathway

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