Central and storage carbon metabolism of the brown alga Ectocarpus siliculosus: insights into the origin and evolution of storage carbohydrates in Eukaryotes

International audienceBrown algae exhibit a unique carbon (C) storage metabolism. The photoassimilate d‐fructose 6‐phosphate is not used to produce sucrose but is converted into d‐mannitol. These seaweeds also store C as β‐1,3‐glucan (laminarin), thus markedly departing from most living organisms, which use α‐1,4‐glucans (glycogen or starch).Using a combination of bioinformatic and phylogenetic approaches, we identified the candidate genes for the enzymes involved in C storage in the genome of the brown alga Ectocarpus siliculosus and traced their evolutionary origins.Ectocarpus possesses a complete set of enzymes for synthesis of mannitol, laminarin and trehalose. By contrast, the pathways for sucrose, starch and glycogen are completely absent.The synthesis of β‐1,3‐glucans appears to be a very ancient eukaryotic pathway. Brown algae inherited the trehalose pathway from the red algal progenitor of phaeoplasts, while the mannitol pathway was acquired by lateral gene transfer from Actinobacteria. The starch metabolism of the red algal endosymbiont was entirely lost in the ancestor of Stramenopiles. In light of these novel findings we question the validity of the ‘Chromalveolate hypothesis’

Chlorophyll-binding proteins revisited - a multigenic family of light-harvesting and stress proteins from a brown algal perspective

<p>Abstract</p> <p>Background</p> <p>Chlorophyll-binding proteins (CBPs) constitute a large family of proteins with diverse functions in both light-harvesting and photoprotection. The evolution of CBPs has been debated, especially with respect to the origin of the LI818 subfamily, members of which function in non-photochemical quenching and have been found in chlorophyll a/c-containing algae and several organisms of the green lineage, but not in red algae so far. The recent publication of the <it>Ectocarpus siliculosus </it>genome represents an opportunity to expand on previous work carried out on the origin and function of CBPs.</p> <p>Results</p> <p>The <it>Ectocarpus </it>genome codes for 53 CBPs falling into all major families except the exclusively green family of chlorophyll a/b binding proteins. Most stress-induced CBPs belong to the LI818 family. However, we highlight a few stress-induced CBPs from <it>Phaeodactylum tricornutum </it>and <it>Chondrus crispus </it>that belong to different sub-families and are promising targets for future functional studies. Three-dimensional modeling of two LI818 proteins revealed features common to all LI818 proteins that are likely to interfere with their capacity to bind chlorophyll b and lutein, but may enable binding of chlorophyll c and fucoxanthin. In the light of this finding, we examined the possibility that LI818 proteins may have originated in a chlorophyll c/fucoxanthin containing organism and compared this scenario to three alternatives: an independent evolution of LI818 proteins in different lineages, an ancient origin together with the first CBPs, before the separation of the red and the green lineage, or an origin in the green lineage and a transfer to an ancestor of haptophytes and heterokonts during a cryptic endosymbiosis event.</p> <p>Conclusions</p> <p>Our findings reinforce the idea that the LI818 family of CBPs has a role in stress response. In addition, statistical analyses of phylogenetic trees show an independent origin in different eukaryotic lineages or a green algal origin of LI818 proteins to be highly unlikely. Instead, our data favor an origin in an ancestral chlorophyll a/c-containing organism and a subsequent lateral transfer to some green algae, although an origin of LI818 proteins in a common ancestor of red and green algae cannot be ruled out.</p

Identification of a very long chain polyunsaturated fatty acid Δ4-desaturase from the microalga Pavlova lutheri11The sequence reported in this paper has been submitted to GenBank database under the accession number AY332747.

AbstractPavlova lutheri, a marine microalga, is rich in the very long chain polyunsaturated fatty acids (VLCPUFAs) eicosapentaenoic (20:5n-3) and docosahexaenoic (22:6n-3) acids. Using an expressed sequence tag approach, we isolated a cDNA designated Pldes1, and encoding an amino acid sequence showing high similarity with polyunsaturated fatty acid front-end desaturases. Heterologous expression in yeast demonstrated that PlDES1 desaturated 22:5n-3 and 22:4n-6 into 22:6n-3 and 22:5n-6 respectively, and was equally active on both substrates. Thus, PlDES1 is a novel VLCPUFA Δ4-desaturase. Pldes1 expression is four-fold higher during the mid-exponential phase of growth compared to late exponential and stationary phases

Inorganic fillers influence on the radiation-induced ageing of a space-used silicone elastomer

A space-used filled silicone rubber (silica and iron oxide fillers) and its polysiloxane isolated matrix were exposed to high energy electrons in order to determine their ageing mechanisms from a structural point of view. Physicochemical analysis evidenced that both filled and unfilled materials predominantly crosslink under such irradiation. Solid-state 29Si NMR spectroscopy allowed the identification of T-type SiO3 units as the main new crosslinks formed in the polymer network. It also revealed an increase in Qtype SiO4 units in the irradiated filled sample. Thanks to the combination of NMR spectroscopy and ammonia-modified swelling tests, these Q-type units were associated with new crosslinks formed at the silica fillers-matrix interface. While the main interaction between the polysiloxane network and the fillers was shown to proceed mainly through hydrogen bonding in the pristine filled samples, it was suggested that the hydrogen bonds were progressively replaced with SiO4 chemical bonds. These additional chemical crosslinks induced evolutions of the shear modulus on the rubber plateau and crosslink density that were significantly more pronounced in the filled material than in the neat one

Electrical conductivity of a silicone network upon electron irradiation: influence of formulation

In this study, the electrical conductivity of a silicone elastomer filled with inorganic fillers was investigated upon electron irradiation. Neat samples consisting of the isolated polysiloxane matrix (with no fillers) were studied in parallel to identify the filler contribution to this evolution. It was shown that exposure to 400 keV electron doses induced a decrease in electrical conductivity for both the filled and neat materials. This decrease was much more pronounced with the filled samples than with the neat ones. Moreover, the activation energy of electrical conductivity (Arrhenius behaviour) doubled in the filled case, while it varied only weakly for the neat case. In light of these results, structure–property relationships were proposed on the basis of the radiation-induced crosslink processes to which this material is subject. In the framework of electronic percolation theory, it is suggested that the radiation-induced formation of SiO3 crosslinks in the polysiloxane network and SiO4 crosslinks at filler–matrix interfaces affects the percolation path of the material, which can be simply modelled by a network of resistors in series. On one hand, their densification increases the overall resistance of the percolation path, which results in the observed decrease of effective electrical conductivity. On the other hand, the steep increase in activation energy in the filled material attributes to the SiO4 crosslinks becoming the most restrictive barrier along the percolation path. In spite of the misleading likeness of electrical conductivities in the pristine state, this study presented evidence that silicone formulation can affect the evolution of electrical properties in radiative environments. To illustrate this conclusion, the use of this material in space applications, especially when directly exposed to the radiative space environment, was discussed. The decrease in electrical conductivity was associated with a progressively increasing risk for the occurrence of electrostatic discharge and consequent spacecraft failures

In silico identification of bacterial seaweed-degrading bioplastic producers

There is an urgent need to replace petroleum-based plastic with bio-based and biodegradable alternatives. Polyhydroxyalkanoates (PHAs) are attractive prospective replacements that exhibit desirable mechanical properties and are recyclable and biodegradable in terrestrial and marine environments. However, the production costs today still limit the economic sustainability of the PHA industry. Seaweed cultivation represents an opportunity for carbon capture, while also supplying a sustainable photosynthetic feedstock for PHA production. We mined existing gene and protein databases to identify bacteria able to grow and produce PHAs using seaweed-derived carbohydrates as substrates. There were no significant relationships between the genes involved in the deconstruction of algae polysaccharides and PHA production, with poor to negative correlations and diffused clustering suggesting evolutionary compartmentalism. We identified 2 987 bacterial candidates spanning 40 taxonomic families predominantly within Alphaproteobacteria, Gammaproteobacteria and Burkholderiales with enriched seaweed-degrading capacity that also harbour PHA synthesis potential. These included highly promising candidates with specialist and generalist specificities, including Alteromonas, Aquisphaera, Azotobacter, Bacillus, Caulobacter, Cellvibrionaceae, Duganella, Janthinobacterium, Massilia, Oxalobacteraceae, Parvularcula, Pirellulaceae, Pseudomonas, Rhizobacter, Rhodanobacter, Simiduia, Sphingobium, Sphingomonadaceae, Sphingomonas, Stieleria, Vibrio and Xanthomonas. In this enriched subset, the family-level densities of genes targeting green macroalgae polysaccharides were considerably higher ( n=231.6±68.5) than enzymes targeting brown ( n=65.34±13.12) and red ( n=30.5±10.72) polysaccharides. Within these organisms, an abundance of FabG genes was observed, suggesting that the fatty acid de novo synthesis pathway supplies (R)-3-hydroxyacyl-CoA or 3-hydroxybutyryl-CoA from core metabolic processes and is the predominant mechanism of PHA production in these organisms. Our results facilitate extending seaweed biomass valorization in the context of consolidated biorefining for the production of bioplastics

Electrical behaviour of a silicone elastomer under simulated space environment

The electrical behavior of a space-used silicone elastomer was characterized using surface potential decay and dynamic dielectric spectroscopy techniques. In both cases, the dielectric manifestation of the glass transition (dipole orientation) and a charge transport phenomenon were observed. An unexpected linear increase of the surface potential with temperature was observed around Tg in thermally-stimulated potential decay experiments, due to molecular mobility limiting dipolar orientation in one hand, and 3D thermal expansion reducing the materials capacitance in the other hand. At higher temperatures, the charge transport process, believed to be thermally activated electron hopping with an activation energy of about 0.4 eV, was studied with and without the silica and iron oxide fillers present in the commercial material. These fillers were found to play a preponderant role in the low-frequency electrical conductivity of this silicone elastomer, probably through a Maxwell–Wagner–Sillars relaxation phenomenon

Engineering mannitol biosynthesis in Escherichia coli and Synechococcus sp. PCC 7002 using a green algal fusion protein

The genetic engineering of microbial cell factories is a sustainable alternative to the chemical synthesis of organic compounds. Successful metabolic engineering often depends on manipulating several enzymes, requiring multiple transformation steps and selection markers, as well as protein assembly and efficient substrate channeling. Naturally occurring fusion genes encoding two or more enzymatic functions may offer an opportunity to simplify the engineering process and to generate ready-made protein modules, but their functionality in heterologous systems remains to be tested. Here we show that heterologous expression of a fusion enzyme from the marine alga Micromonas pusilla, comprising a mannitol-1-phosphate dehydrogenase and a mannitol-1-phosphatase, leads to synthesis of mannitol by Escherichia coli and by the cyanobacterium Synechococcus sp. PCC 7002. Neither of the heterologous systems naturally produces this sugar alcohol, which is widely used in food, pharmaceutical, medical and chemical industries. While the mannitol production rates obtained by single-gene manipulation were lower than those previously achieved after pathway optimization with multiple genes, our findings show that naturally occurring fusion proteins can offer simple building blocks for the assembly and optimization of recombinant metabolic pathways