Production of taxa-4(5),11(12)-diene by transgenic Physcomitrella patens

Anterola, Aldwin; Shanle, Erin K.; Perroud, Pierre‐François; Quatrano, Ralph S.

doi:10.1007/s11248-009-9252-5

Cited by 80 publications

(49 citation statements)

References 40 publications

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“…Some photosynthetic hosts such as cyanobacteria and eukaryotic moss tissue culture have already been demonstrated to be amenable to heterologous terpenoid expression (Anterola et al, 2009;Davies et al, 2014;Halfmann et al, 2014;Zhan et al, 2014). Eukaryotic microalgae as photosynthetic production platforms, however, are notoriously difficult to genetically engineer and there are limited reports on synthetic metabolic modifications of these organisms (Cordero et al, 2011;Kaye et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii

Lauersen

Baier

Wichmann

et al. 2016

Metabolic Engineering

118

231

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Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii, Metabolic Engineering, http://dx.doi.org/10.1016/j.ymben.2016 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting galley proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. olaf.kruse@uni-bielefeld.de AbstractThe heterologous expression of terpene synthases in microbial hosts has opened numerous possibilities for bioproduction of desirable metabolites. Photosynthetic microbial hosts present a sustainable alternative to traditional fermentative systems, using freely available (sun)light and carbon dioxide as inputs for bio-production. Here, we report the expression of a patchoulol synthase from Pogostemon cablin Benth in the model green microalga Chlamydomonas reinhardtii. The sesquiterpenoid patchoulol was produced from the alga and was used as a marker of sesquiterpenoid production capacity. A novel strategy for gene loading was employed and patchoulol was produced up to 922 ± 242 µg g -1 CDW in six days. We additionally investigated the effect of carbon source on sesquiterpenoid productivity from C. reinhardtii in scale-up batch cultivations. It was determined that up to 1.03 mg L -1 sesquiterpenoid products could be produced in completely photoautotrophic conditions and that the alga exhibited altered sesquiterpenoid production metabolism related to carbon source.

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Section: Introductionmentioning

confidence: 99%

Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii

Lauersen

Baier

Wichmann

et al. 2016

Metabolic Engineering

118

231

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“…The presence and expression of the TS gene in the transgenic cultured ginseng root and the de novo formation of a compound with the same fragmentation pattern as taxadiene confirmed the successful transformation of cultured ginseng roots to produce taxadiene. The taxadiene yield from the transformed ginseng roots with the TS gene was 9.10 μg/g dry weight, which was higher than the 5 μg/g dry weight produced in moss (30), but lower than the 20 μg/g dry weight produced in transgenic tomato fruits (29).…”

Section: Discussionmentioning

confidence: 70%

“…Several studies have indicated methods to taxadiene in tomato and Arabidopsis using metabolic engineering due to the high price (29)(30)(31). The presence and expression of the TS gene in the transgenic cultured ginseng root and the de novo formation of a compound with the same fragmentation pattern as taxadiene confirmed the successful transformation of cultured ginseng roots to produce taxadiene.…”

Section: Discussionmentioning

confidence: 95%

Production of taxadiene from cultured ginseng roots transformed with taxadiene synthase gene

Cha¹,

Shim²,

Kim³

et al. 2012

BMB Reports

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Paclitaxel is produced by various species of yew trees and has been extensively used to treat tumors. In our research, a taxadiene synthase (TS) gene from Taxus brevifolia was used to transform the roots of cultured ginseng (Panax ginseng C.A. Meyer) to produce taxadiene, the unique skeletal precursor to taxol. The TS gene was successfully introduced into the ginseng genome, and the de novo formation of taxadiene was identified by mass spectroscopy profiling. Without any change in phenotypes or growth difference in a TS-transgenic ginseng line, the transgenic TSS3-2 line accumulated 9.1 μg taxadiene per gram of dry weight. In response to the treatment of methyl jasmonate for 3 or 6 days, the accumulation was 14.6 and 15.9 μg per g of dry weight, respectively. This is the first report of the production of taxadiene by engineering ginseng roots with a taxadiene synthase gene. [BMB Reports 2012; 45(10): 589-594]

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“…35) P. patens was transfected with the biosynthetic gene for taxol and was confirmed to produce the biosynthetic intermediate. 36) Thus, these taxol-accumulating species including T. cuspidata, would be characterized as having the D26E mutation in common.…”

Section: Discussionmentioning

confidence: 99%

Expression of recombinant alpha and beta tubulins from the yew Taxus cuspidata and analysis of the microtubule assembly in the presence of taxol

Kudo

Abe

Ito

et al. 2014

Bioscience, Biotechnology, and Biochemistry

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Taxol was originally isolated from the yew Taxus brevifolia. Because taxol inhibits the depolymerization of microtubules, the presence of a self-resistance mechanism in Taxus spp. was hypothesized. The cloning of the cDNA for alpha and beta tubulins from Taxus cuspidata and those from the human embryonic kidney cell line HEK293T revealed that the 26Asp, 359Arg, and 361Leu residues in the human beta tubulin, which are important for taxol binding, were replaced with Glu, Trp, and Met in the beta tubulin of T. cuspidata, respectively. The microtubule assembly of the recombinant alpha and beta tubulins was monitored turbidimetrically, and the results clearly demonstrated that the microtubule from T. cuspidata is less sensitive to taxol than that from HEK293T cells. The Taxus microtubule composed of the wild-type alpha tubulin and the beta tubulin with the E26D mutation restored the sensitivity to taxol. We thus postulated that the mutation identified in the beta tubulin of T. cuspidata plays a role in the self-resistance of this species against taxol.

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Production of taxa-4(5),11(12)-diene by transgenic Physcomitrella patens

Cited by 80 publications

References 40 publications

Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii

Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii

Production of taxadiene from cultured ginseng roots transformed with taxadiene synthase gene

Expression of recombinant alpha and beta tubulins from the yew Taxus cuspidata and analysis of the microtubule assembly in the presence of taxol

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