Novel FK506 and FK520 analogues <i>via</i> mutasynthesis: mutasynthon scope and product characteristics

Moss, Steven J.; Stanley-Smith, Anna E.; Schell, Ursula; Coates, Nigel; Foster, Teresa; Gaisser, Sibylle; Gregory, Matthew A.; Martin, Christine; Nur‐e‐Alam, Mohammad; Piraee, Mahmood; Radzom, Markus; Suthar, Dipen; Thexton, David G.; Warneck, Tony; Zhang, Ming‐Qiang; Wilkinson, Barrie

doi:10.1039/c2md20266b

Cited by 13 publications

(13 citation statements)

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“…Genetic manipulation of these pathways has been successfully applied to produce useful libraries of bacterial complex polyketides, such as the immunosuppressants rapamycin[ 4 ] and FK506. [ 5 ] However, extension of this attractive engineering approach to many other such natural products has not proved routine, because the limits to the modularity of the synthases and the specificity of auxiliary enzymes are poorly understood.…”

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confidence: 99%

Site‐Specific Modification of the Anticancer and Antituberculosis Polyether Salinomycin by Biosynthetic Engineering

et al. 2014

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The complex bis-spiroacetal polyether ionophore salinomycin has been identified as a uniquely selective agent against cancer stem cells and is also strikingly effective in an animal model of latent tuberculosis. The basis for these important activities is unknown. We show here that deletion of the salE gene abolishes salinomycin production and yields two new analogues, in both of which the C18=C19 cis double bond is replaced by a hydroxy group stereospecifically located at C19, but which differ from each other in the configuration of the bis-spiroacetal. These results identify SalE as a novel dehydratase and demonstrate that biosynthetic engineering can be used to redirect the reaction cascade of oxidative cyclization to yield new salinomycin analogues for use in mechanism-of-action studies.

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confidence: 99%

Site‐Specific Modification of the Anticancer and Antituberculosis Polyether Salinomycin by Biosynthetic Engineering

et al. 2014

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“…Thus, in addition to identifying the described changes, we tested several media for their suitability for plating of the conjugation mixture of E. coli cells and Streptomyces spores in place of MS medium, as described in reference 23. Replacement of the MS medium with R6 agar containing 48 mM CaCl 2 (a medium suitable for killing E. coli [16,48]) gave stable exconjugants from each postconjugation plate.…”

Section: Resultsmentioning

confidence: 99%

An Efficient Method To Generate Gene Deletion Mutants of the Rapamycin-Producing Bacterium Streptomyces iranensis HM 35

Netzker

Schroeckh

Gregory

et al. 2016

Appl Environ Microbiol

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Streptomyces iranensis HM 35 is an alternative rapamycin producer to Streptomyces rapamycinicus. Targeted genetic modification of rapamycin-producing actinomycetes is a powerful tool for the directed production of rapamycin derivatives, and it has also revealed some key features of the molecular biology of rapamycin formation in S. rapamycinicus. The approach depends upon efficient conjugational plasmid transfer from Escherichia coli to Streptomyces, and the failure of this step has frustrated its application to Streptomyces iranensis HM 35. Here, by systematically optimizing the process of conjugational plasmid transfer, including screening of various media, and by defining optimal temperatures and concentrations of antibiotics and Ca 2؉ ions in the conjugation media, we have achieved exconjugant formation for each of a series of gene deletions in S. iranensis HM 35. Among them were rapK, which generates the starter unit for rapamycin biosynthesis, and hutF, encoding a histidine catabolizing enzyme. The protocol that we have developed may allow efficient generation of targeted gene knockout mutants of Streptomyces species that are genetically difficult to manipulate. IMPORTANCE The developed protocol of conjugational plasmid transfer from Escherichia coli to Streptomyces iranensis may allow efficient generation of targeted gene knockout mutants of other genetically difficult to manipulate, but valuable, Streptomyces species. Since the discovery of streptomycin in 1943 (1), streptomycetes have been shown to produce thousands of compounds with possibly beneficial features, e.g., antibiotics, immunosuppressants, or anticancer drugs. Actinomycete-derived metabolites comprise over two-thirds of all known antibiotic compounds (2), and recent genome sequencing programs revealed that their biosynthesis potential has been underestimated. In their 8-to 12-Mb genomes, approximately 20 to 30 gene clusters encode the biosynthesis of secondary metabolites (3-5). One of the most important Streptomyces-derived compounds is the mechanistic target of rapamycin (mTOR) inhibitor rapamycin (sirolimus), a "billion dollar molecule" and, after cyclosporine, the most widely used immunosuppressant of microbial origin (6). Rapamycin was previously reported as an antifungal antibiotic produced by Streptomyces hygroscopicus ATCC 29253 (7), later renamed Streptomyces rapamycinicus (8). The rapamycin gene cluster in this strain has been sequenced (9), the biosynthetic pathway has been extensively characterized (10-14), and engineering of the cluster has yielded an impressive range of bioactive modified rapamycins (rapalogs) (15, 16), some in multigram amounts. So far, two other rapamycin-producing species are known: the taxonomically closely related Streptomyces iranensis HM 35 (5, 17) and Actinoplanes sp. strain N902-109 (18). To elucidate the molecular biology of rapamycin formation in these strains and to further exploit the physiological and pharmacological capability of rapamycin derivatives, genetic manipulation of these altern...

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“…The full‐length chain then undergoes further chemical transformation by additional enzymes. Genetic manipulation of these pathways has been successfully applied to produce useful libraries of bacterial complex polyketides, such as the immunosuppressants rapamycin4 and FK506 5. However, extension of this attractive engineering approach to many other such natural products has not proved routine, because the limits to the modularity of the synthases and the specificity of auxiliary enzymes are poorly understood.…”

Section: Methodsmentioning

confidence: 99%

High‐Performance Polymers from Nature: Catalytic Routes and Processes for Industry

Walther

2014

ChemSusChem

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It is difficult to imagine life today without polymers. However, most chemicals are almost exclusively synthesized from petroleum. With diminishing oil reserves, establishing an industrial process to transform renewables into high-value chemicals may be more challenging than running a car without gasoline. This is due to the difficulty in setting up processes that are novel, profitable, and environmentally benign at the same time. Additionally, the quest for sustainability of renewable resources should be based on incorporating ethical considerations in the development of plans that utilize feedstocks intended for human nutrition and health. Thus, it is important to use bio-energy containing renewable resources in the most efficient way. This Concept goes beyond the synthesis of monomers and provides insights for establishing an industrial process that transforms renewable resources into high-value chemicals, and it describes careful investigations that are of paramount importance, including evaluations from an economical and an ecological perspective. The synthesis of monomers suitable for polymer production from renewable resources would ideally be accompanied by a reduction in CO2 emission and waste, through the complete molecular utilization of the feedstock. This Concept advocates the drop-in strategy, and is guided by the example of catalytically synthesized dimethyl 1,19-nonadecanedioate and its α,ω-functionalized derivatives. With respect to the Twelve Principles of Green Chemistry, this Concept describes a technological leap forward for a sustainable green chemical industry.

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Novel FK506 and FK520 analogues via mutasynthesis: mutasynthon scope and product characteristics

Abstract: Novel FK506 and FK520 analogues were generated via biosynthetic engineering in order to generate analogue compounds with equal potency but improved pharmacological profiles compared to FK506.

Cited by 13 publications

References 52 publications

Site‐Specific Modification of the Anticancer and Antituberculosis Polyether Salinomycin by Biosynthetic Engineering

Site‐Specific Modification of the Anticancer and Antituberculosis Polyether Salinomycin by Biosynthetic Engineering

An Efficient Method To Generate Gene Deletion Mutants of the Rapamycin-Producing Bacterium Streptomyces iranensis HM 35

High‐Performance Polymers from Nature: Catalytic Routes and Processes for Industry

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