Peroxisomal microbodies are at the crossroads of acetate assimilation in the green microalga Chlamydomonas reinhardtii

Lauersen, Kyle J.; Willamme, Rémi; Coosemans, Nadine; Joris, Marine; Kruse, Olaf; Remacle, Claire

doi:10.1016/j.algal.2016.03.026

Cited by 55 publications

(88 citation statements)

References 43 publications

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“…C. reinhardtii lacks the MVA pathway, therefore, the FPP pool must be generated entirely from IPP and DMAPP produced in the chloroplast (Lohr et al, 2012). When acetate is fed to the cells, the main carbon metabolism is dependent on the ATP-requiring process of acetate uptake and conversion via the glyoxylate cycle into C4 components for cellular building blocks (Lauersen et al, 2016). Although ATP is generated by light and electron flow through photosystems in the chloroplasts, it is likely that the cells produce more FPP as a UQ precursor to drive aerobic respiration processes under ATP requiring, acetate consuming and low CO 2 conditions.…”

Section: Increasing Sesquiterpene Synthase Titre Can Increase Relativmentioning

confidence: 99%

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

Lauersen

Baier

Wichmann

et al. 2016

Metabolic Engineering

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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: Increasing Sesquiterpene Synthase Titre Can Increase Relativmentioning

confidence: 99%

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

Lauersen

Baier

Wichmann

et al. 2016

Metabolic Engineering

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120

231

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“…reinhardtii also can grow mixotrophically using alternative organic carbon sources present in its environment. For example, it can take up acetate, which is then incorporated into the citric cycle, producing reducing equivalents and CO 2 (Johnson and Alric, 2012), and into the glyoxylate cycle, producing malate (Lauersen et al, 2016). In the presence of acetate, it has been reported that CO 2 uptake and oxygen evolution were decreased by half under saturating CO 2 and light intensities without affecting PSII efficiency, respiration, and cell growth (Heifetz et al, 2000).…”

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confidence: 99%

Carbon Supply and Photoacclimation Cross Talk in the Green Alga Chlamydomonas reinhardtii

et al. 2016

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Photosynthetic organisms are exposed to drastic changes in light conditions, which can affect their photosynthetic efficiency and induce photodamage. To face these changes, they have developed a series of acclimation mechanisms. In this work, we have studied the acclimation strategies of Chlamydomonas reinhardtii, a model green alga that can grow using various carbon sources and is thus an excellent system in which to study photosynthesis. Like other photosynthetic algae, it has evolved inducible mechanisms to adapt to conditions where carbon supply is limiting. We have analyzed how the carbon availability influences the composition and organization of the photosynthetic apparatus and the capacity of the cells to acclimate to different light conditions. Using electron microscopy, biochemical, and fluorescence measurements, we show that differences in CO 2 availability not only have a strong effect on the induction of the carbon-concentrating mechanisms but also change the acclimation strategy of the cells to light. For example, while cells in limiting CO 2 maintain a large antenna even in high light and switch on energy-dissipative mechanisms, cells in high CO 2 reduce the amount of pigments per cell and the antenna size. Our results show the high plasticity of the photosynthetic apparatus of C. reinhardtii. This alga is able to use various photoacclimation strategies, and the choice of which to activate strongly depends on the carbon availability.

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“… In vivo Cr GFY1–5 protein localization and immunoblotting analyses. (A) Schematic overview of vectors used for Cr GFY‐fluorescent reporter expression in this study: ( a ) cytosolic YFP and ( b ) peroxisomal YFP containing the PTS1 corresponding to the MAS1_PTS1 malate synthase C‐terminal peptide of C. reinhardtii (HIVTKTPSRM*) (Lauersen, Willamme, et al, ); ( c ) cytosolic CFP and ( d ) CFP carrying 20 amino acid secretion signal from Cr CAH1 enzyme ( c CA) and the KDEL* peptide at its C‐terminus for ER retention. Two gray arrows represent the hybrid promoter HSP70‐RBCS2; a gray box depicts the RBCS2 3′UTR; selection markers conferring resistance to hygromycin B (H) or paromomycin (P) are indicated; RBCS2 introns 1 and 2 of the pOpt vectors are depicted from left to right by thin bridge lines in vectors a–d.…”

Section: Resultsmentioning

confidence: 99%

“…Vectors a and c are pOpt_mVenus_Paro and pOpt_mCerulean3_Hyg, respectively, from Lauersen et al., . Vector b was previously developed by our group (Lauersen, Willamme, et al., ). This vector expresses a modified mVenus (YFP) reporter with the C‐terminal amino acids of C. reinhardtii malate synthase 1 gene HIVTKTPSRM*, which is a confirmed peroxisomal targeting signal type 1 (PTS1) sequence (Lauersen, Willamme, et al., ).…”

Section: Methodsmentioning

confidence: 99%

“…In eukaryotic cells, the enzymatic reactions of glyoxylate cycle are distributed between the peroxisomal matrix and the cytosol (Kunze & Hartig, ; Kunze, Pracharoenwattana, Smith, & Hartig, ). All enzymes of the glyoxylate cycle in C. reinhardtii have been localized to algal peroxisomes including a single acetyl‐CoA synthase (ACS3), with the sole exception of ICL1, which is cytosolic (Lauersen, Willamme, et al., ). A mutant deficient in ICL1 was unable to utilize acetate heterotrophically and exhibited reduced enzyme abundances of other glyoxylate cycle genes (Plancke et al., ).…”

Section: Introductionmentioning

confidence: 99%

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Characterization of the GPR1/FUN34/YaaH protein family in the green microalga Chlamydomonas suggests their role as intracellular membrane acetate channels

et al. 2019

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The unicellular green microalga Chlamydomonas reinhardtii is a powerful photosynthetic model organism which is capable of heterotrophic growth on acetate as a sole carbon source. This capacity has enabled its use for investigations of perturbations in photosynthetic machinery as mutants can be recovered heterotrophically. Fixation of acetate into cellular carbon metabolism occurs first by its conversion into acetyl‐CoA by a respective synthase and the generation of succinate by the glyoxylate cycle. These metabolic steps have been recently determined to largely occur in the peroxisomes of this alga; however, little is known about the trafficking and import of acetate or its subcellular compartmentalization. Recently, the genes of five proteins belonging to the GPR 1/ FUN 34/YaaH ( GFY ) superfamily were observed to exhibit increased expression in C. reinhardtii upon acetate addition, however, no further characterization has been reported. Here, we provide several lines of evidence to implicate Cr GFY 1–5 as channels which share structural homology with bacterial succinate‐acetate channels and specifically localize to microbodies, which are surprisingly distinct from the glyoxylate cycle‐containing peroxisomes. We demonstrate structural models, gene expression profiling, and in vivo fluorescence localization of all five isoforms in the algal cell to further support this role.

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Peroxisomal microbodies are at the crossroads of acetate assimilation in the green microalga Chlamydomonas reinhardtii

Cited by 55 publications

References 43 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

Carbon Supply and Photoacclimation Cross Talk in the Green Alga Chlamydomonas reinhardtii

Characterization of the GPR1/FUN34/YaaH protein family in the green microalga Chlamydomonas suggests their role as intracellular membrane acetate channels

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