Expanding the Landscape of Diterpene Structural Diversity through Stereochemically Controlled Combinatorial Biosynthesis

Andersen-Ranberg, Johan; Kongstad, Kenneth T.; Nielsen, Morten Ørregaard; Jensen, Niels Bjerg; Pateraki, Irini; Bach, Søren Spanner; Hamberger, Britta; Zerbe, Philipp; Bohlmann, Jörg; Møller, Birger Lindberg; Hamberger, Björn

doi:10.1002/ange.201510650

Cited by 41 publications

(72 citation statements)

References 27 publications

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“…To test the capacity of the identified P450s to oxygenate casbene, E. lathyris CYP71D445 and CYP726A27 were coexpressed with CBS in N. benthamiana using the Agrobacterium-mediated transient expression system, engineered for optimized production of diterpenoids by the strategy of coexpression with Coleus forskohlii 1-deoxy-D-xylulose 5-phosphate synthase (DXS) and geranylgeranyl diphosphate synthase (GGPPS) as described earlier (23). The production of casbene (1) in the infiltrated N. benthamiana leaves was confirmed by GC-MS analysis ( Fig.…”

Section: Ms (Lc-hrms)mentioning

confidence: 99%

Oxidation and cyclization of casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L.

Luo

Callari

Hamberger

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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The seed oil of Euphorbia lathyris L. contains a series of macrocyclic diterpenoids known as Euphorbia factors. They are the current industrial source of ingenol mebutate, which is approved for the treatment of actinic keratosis, a precancerous skin condition. Here, we report an alcohol dehydrogenase-mediated cyclization step in the biosynthetic pathway of Euphorbia factors, illustrating the origin of the intramolecular carbon-carbon bonds present in lathyrane and ingenane diterpenoids. This unconventional cyclization describes the ring closure of the macrocyclic diterpene casbene. Through transcriptomic analysis of E. lathyris L. mature seeds and in planta functional characterization, we identified three enzymes involved in the cyclization route from casbene to jolkinol C, a lathyrane diterpene. These enzymes include two cytochromes P450 from the CYP71 clan and an alcohol dehydrogenase (ADH). CYP71D445 and CYP726A27 catalyze regio-specific 9-oxidation and 5-oxidation of casbene, respectively. When coupled with these P450-catalyzed monooxygenations, E. lathyris ADH1 catalyzes dehydrogenation of the hydroxyl groups, leading to the subsequent rearrangement and cyclization. The discovery of this nonconventional cyclization may provide the key link to complete elucidation of the biosynthetic pathways of ingenol mebutate and other bioactive macrocyclic diterpenoids.terpenoid biosynthesis | transcriptomic analysis | regio-specific oxidation | nonconventional cyclization

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Section: Ms (Lc-hrms)mentioning

confidence: 99%

Oxidation and cyclization of casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L.

Luo

Callari

Hamberger

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…It is interesting to note that the recent emergence enzymes MvELS in M. vulgare (12.6 Ma; Roy and Lindqvist, 2015) and SsScS in S. sclarea (10 Ma; Drew and Sytsma, 2012) exhibit more diversified enzymatic activity than other KSLs Schalk et al, 2012;Andersen-Ranberg et al, 2016;Jia et al, 2016). This alludes to the fast evolution of KSL enzymes in Lamiaceae.…”

Section: The Evolution Of Domain Loss In Lamiaceaementioning

confidence: 96%

“…Many of these enzymes have lost the characteristic internal/g domain that is responsible for metabolite specialization, a deletion event found in KSLs in Lamiaceae (Hillwig et al, 2011;Caniard et al, 2012;Schalk et al, 2012;Brückner et al, 2014;Pateraki et al, 2014;Zerbe et al, 2014;Cui et al, 2015;Bo zi c et al, 2015;Trikka et al, 2015). A recent work has shown the plasticity of class I KSLs, such as SsScS from S. sclarea, that can react with any available CPS product (Andersen-Ranberg et al, 2016;Jia et al, 2016). These studies demonstrate the value of Lamiaceae as a useful system to study diterpenoid chemical diversity evolution.…”

mentioning

confidence: 99%

Functional Diversification of Kaurene Synthase-Like Genes in Isodon rubescens

et al. 2017

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“…Four industrially relevant "targets" were achieved in scalable biotechnological production of therapeutically important diterpenoids involving the use of bioengineered strains of S. cerevisiae as cell factories. [42] Bioengineered wine yeast uncorks a future with synthetic DNA History tells us that wine yeast strain development is well positioned to, once again, benefit from technological advances made with the genetic and genome engineering of non-wine strains of S. cerevisiae. To date, using genetic engineering [20,43,44] and non-GM techniques, [45] such as classical breeding (hybridization), adaptive laboratory evolution, and mutagenesis, several non-GM and GM wine strains have been developed with increased robustness, fermentation performance, health-related properties, and/or sensory attributes.…”

Section: Bioengineered Yeast Put Synthetic Dna To Work In Industrymentioning

confidence: 99%

Synthetic genome engineering forging new frontiers for wine yeast

Pretorius

2016

Critical Reviews in Biotechnology

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Over the past 15 years, the seismic shifts caused by the convergence of biomolecular, chemical, physical, mathematical, and computational sciences alongside cutting-edge developments in information technology and engineering have erupted into a new field of scientific endeavor dubbed Synthetic Biology. Recent rapid advances in high-throughput DNA sequencing and DNA synthesis techniques are enabling the design and construction of new biological parts (genes), devices (gene networks) and modules (biosynthetic pathways), and the redesign of biological systems (cells and organisms) for useful purposes. In 2014, the budding yeast Saccharomyces cerevisiae became the first eukaryotic cell to be equipped with a fully functional synthetic chromosome. This was achieved following the synthesis of the first viral (poliovirus in 2002 and bacteriophage Phi-X174 in 2003) and bacterial (Mycoplasma genitalium in 2008 and Mycoplasma mycoides in 2010) genomes, and less than two decades after revealing the full genome sequence of a laboratory (S288c in 1996) and wine (AWRI1631 in 2008) yeast strain. A large international project -the Synthetic Yeast Genome (Sc2.0) Project -is now underway to synthesize all 16 chromosomes ($12 Mb carrying $6000 genes) of the sequenced S288c laboratory strain by 2018. If successful, S. cerevisiae will become the first eukaryote to cross the horizon of in silico design of complex cells through de novo synthesis, reshuffling, and editing of genomes. In the meantime, yeasts are being used as cell factories for the semi-synthetic production of high-value compounds, such as the potent antimalarial artemisinin, and food ingredients, such as resveratrol, vanillin, stevia, nootkatone, and saffron. As a continuum of previously genetically engineered industrially important yeast strains, precision genome engineering is bound to also impact the study and development of wine yeast strains supercharged with synthetic DNA. The first taste of what the future holds is the de novo production of the raspberry ketone aroma compound, 4-[4-hydroxyphenyl]butan-2-one, in a wine yeast strain (AWRI1631), which was recently achieved via metabolic pathway engineering and synthetic enzyme fusion. A peek over the horizon is revealing that the future of "Wine Yeast 2.0" is already here. Therefore, this article seeks to help prepare the wine industry -an industry rich in history and tradition on the one hand, and innovation on the other -for the inevitable intersection of the ancient art practiced by winemakers and the inventive science of pioneering "synthetic genomicists". It would be prudent to proactively engage all stakeholders -researchers, industry practitioners, policymakers, regulators, commentators, and consumers -in a meaningful dialog about the potential challenges and opportunities emanating from Synthetic Biology. To capitalize on the new vistas of synthetic yeast genomics, this paper presents wine yeast research in a fresh context, raises important questions and proposes new directions. ARTICLE HISTORY

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Expanding the Landscape of Diterpene Structural Diversity through Stereochemically Controlled Combinatorial Biosynthesis

Cited by 41 publications

References 27 publications

Oxidation and cyclization of casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L.

Oxidation and cyclization of casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L.

Functional Diversification of Kaurene Synthase-Like Genes in Isodon rubescens

Synthetic genome engineering forging new frontiers for wine yeast

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