Targeted Engineering of Cyclooctat‐9‐en‐7‐ol Synthase: A Stereospecific Access to Two New Non‐natural Fusicoccane‐Type Diterpenes

Görner, Christian; Häuslein, Ina; Schrepfer, Patrick; Eisenreich, Wolfgang; Brück, Thomas

doi:10.1002/cctc.201300285

Cited by 30 publications

(52 citation statements)

References 52 publications

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“… Hypothetical mechanism originally proposed for the formation of cyclooctat‐9‐en‐7‐ol ( 2 ) from GGDP ( 1 ) catalyzed by CotB2 2. 4, 5 OPP=pyrophosphate. …”

Section: Methodsmentioning

confidence: 99%

An Unusual Terpene Cyclization Mechanism Involving a Carbon–Carbon Bond Rearrangement

et al. 2015

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Terpene cyclization reactions are fascinating owing to the precise control of connectivity and stereochemistry during the catalytic process. Cyclooctat-9-en-7-ol synthase (CotB2) synthesizes an unusual 5-8-5 fused-ring structure with six chiral centers from the universal diterpene precursor, the achiral C20 geranylgeranyl diphosphate substrate. An unusual new mechanism for the exquisite CotB2-catalyzed cyclization that involves a carbon-carbon backbone rearrangement and three long-range hydride shifts is proposed, based on a powerful combination of in vivo studies using uniformly (13)C-labeled glucose and in vitro reactions of regiospecifically deuterium-substituted geranylgeranyl diphosphate substrates. This study shows that CotB2 elegantly demonstrates the synthetic virtuosity and stereochemical control that evolution has conferred on terpene synthases.

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“… Hypothetical mechanism originally proposed for the formation of cyclooctat‐9‐en‐7‐ol ( 2 ) from GGDP ( 1 ) catalyzed by CotB2 2. 4, 5 OPP=pyrophosphate. …”

Section: Methodsmentioning

confidence: 99%

An Unusual Terpene Cyclization Mechanism Involving a Carbon–Carbon Bond Rearrangement

et al. 2015

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“…Consistent with this proposed mechanism, two aberrant products, cyclooctat-1,7-diene (6) and cyclooctat-7-en-3-ol (7), have been shown to be formed by the action of the CotB2 mutant enzymes CotB2 F107Y and CotB2 F149L , respectively. [4] The additional derailment products (R)-cembrane A and (1R,3E,7E,11S,12S)-3,7,18-dolabellatriene have been shown to be synthesized by the CotB2 F107A and CotB2 W288G mutants, respectively. [5] To date, however, no direct experimental evidence has been provided to validate the proposed mechanism for the CotB2-catalyzed cyclization reaction.…”

Section: In Memory Of Haruo Setomentioning

confidence: 99%

“…Hypothetical mechanism originallyp roposed for the formation of cyclooctat-9-en-7-ol (2)f rom GGDP (1)catalyzed by CotB2. [2,4,5] OPP = pyrophosphate. in many different ways to yield alarge variety of structurally diverse diterpenes with aw ide spectrum of biological activities that have been isolated from ab road range of natural sources.…”

Section: In Memory Of Haruo Setomentioning

confidence: 99%

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An Unusual Terpene Cyclization Mechanism Involving a Carbon–Carbon Bond Rearrangement

et al. 2015

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Terpene cyclization reactions are fascinating owing to the precise control of connectivity and stereochemistry during the catalytic process. Cyclooctat-9-en-7-ol synthase (CotB2) synthesizes an unusual 5-8-5 fused-ring structure with six chiral centers from the universal diterpene precursor, the achiral C 20 geranylgeranyl diphosphate substrate. An unusual new mechanism for the exquisite CotB2-catalyzed cyclization that involves a carbon-carbon backbone rearrangement and three long-range hydride shifts is proposed, based on a powerful combination of in vivo studies using uniformly 13 C-labeled glucose and in vitro reactions of regiospecifically deuteriumsubstituted geranylgeranyl diphosphate substrates. This study shows that CotB2 elegantly demonstrates the synthetic virtuosity and stereochemical control that evolution has conferred on terpene synthases.

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“…This has also been demonstrated for the bacterial diterpene cyclooctat-9-en-7-ol synthase (CotB2) [57], also a putative Class I TPS. Mutation of tryptophan 288 to glycine in CotB2 resulted in the stereoselective synthesis of (1 R ,3 E ,7 E ,11 S ,12 S )-3,7,18-dolabellatriene, a bicyclic diterpene (Scheme 2).…”

Section: Reviewmentioning

confidence: 65%

Opportunities and challenges for the sustainable production of structurally complex diterpenoids in recombinant microbial systems

Kemper

Hirte

Reinbold

et al. 2017

Beilstein J. Org. Chem.

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With over 50.000 identified compounds terpenes are the largest and most structurally diverse group of natural products. They are ubiquitous in bacteria, plants, animals and fungi, conducting several biological functions such as cell wall components or defense mechanisms. Industrial applications entail among others pharmaceuticals, food additives, vitamins, fragrances, fuels and fuel additives. Central building blocks of all terpenes are the isoprenoid compounds isopentenyl diphosphate and dimethylallyl diphosphate. Bacteria like Escherichia coli harbor a native metabolic pathway for these isoprenoids that is quite amenable for genetic engineering. Together with recombinant terpene biosynthesis modules, they are very suitable hosts for heterologous production of high value terpenes. Yet, in contrast to the number of extracted and characterized terpenes, little is known about the specific biosynthetic enzymes that are involved especially in the formation of highly functionalized compounds. Novel approaches discussed in this review include metabolic engineering as well as site-directed mutagenesis to expand the natural terpene landscape. Focusing mainly on the validation of successful integration of engineered biosynthetic pathways into optimized terpene producing Escherichia coli, this review shall give an insight in recent progresses regarding manipulation of mostly diterpene synthases.

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Targeted Engineering of Cyclooctat‐9‐en‐7‐ol Synthase: A Stereospecific Access to Two New Non‐natural Fusicoccane‐Type Diterpenes

Cited by 30 publications

References 52 publications

An Unusual Terpene Cyclization Mechanism Involving a Carbon–Carbon Bond Rearrangement

An Unusual Terpene Cyclization Mechanism Involving a Carbon–Carbon Bond Rearrangement

An Unusual Terpene Cyclization Mechanism Involving a Carbon–Carbon Bond Rearrangement

Opportunities and challenges for the sustainable production of structurally complex diterpenoids in recombinant microbial systems

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