Konstanz, University, Diss., 2004.

Considering the nature of increasing global temperatures associated with elevated atmospheric carbon dioxide levels as a result of increased demand for energy, it is notable to consider viable options to reduce the strain that these increased carbon dioxide emissions are having on the overall impact of the global climate. Phaeodactylum tricornutum, a marine phytoplankton may be utilized to this end. Its unique ability to increase lipid production under environmental stress conditions, in particular those lipids that can easily be converted into biodiesel, make it an ideal candidate for this use. Here, we examine the effects of trans trans 2,4 decadienal (or DD for short), an aldehyde that is known to induce cell death in the diatom at high concentrations, as they relate to changes in the lipid biosynthesis pathway. 100 ml Axenic cultures of the diatom P. tricornutum were grown to exponential stage, harvested and treated with decadienal at a concentration of 5[mu]g/ml to determine effects on lipid production after 24 hours. Qualitative analysis undertaken using Nile red staining of treated and untreated cells indicated increased fluorescence of treated cells compared to unstained water controls, however this increase may not be attributable to increased lipid production due to the fact that cells were unfixed and must be verified through other means. Initial attempts to verify this finding through thin layer chromatography and qPCR were inconclusive. / text

Através de bioensaios estáticos fo-ram comparadas as taxas máximas de crescimento, razão de fotossintese em termos de clorofila-a e de célula, bem como o rendimento final de tres clones de Phaeodacty-tum tricornutum visando verificar uma possível existência de diferenças intraespe-cificas entre eles. Dois desses clones foram isolados a partir de amostras de Uba-tuba (U3, isolado em 1972 e U5 isolado em 1979) e um de Cananéia (Cl, isolado em 1980). Todos os resultados foram submeti-dos a uma análise de variância trifatorial e a uma anállse de agrupamento com a finlidade de discriminar as condições experimentais nas quais os tres clones diferi-ram significativamente em suas respostas fisiológicas. Durante a fase exponencial de crescimento os tres clones se apresen-taram com morfótipos fusiformes raramente se encontrando morfótipos ovóides e trir-radiados. O padrão das curvas de cresci -mento dos tres clones em diferentes temperaturas (16, 20, 240 C) e meios de cultu-ra (ASP 2, Erd-Schreiber, Guillard fl) não diferiu. Os resultados obtidos indicaram a existência de diferenças significativas nas taxas de crescimento e produti-vidade primária. Morfótipo ovóides for-mando grumos foram observados nos tres clones após a fase exponencial e de modo conspícuo no clone U5. Nas mesmas condi-ções, o clone Cl apresentou uma grande quantidade de grumos, com a forma de uma estrela, constituidos por morfótipos fusiformes unidos convergentemente por uma das extremidades de cada célula. Por outro lado, morfótipos trirradiados e cru-ciformes bem como formação de cadeias, só foram observados no clone U3. / A comparison of the maximum growth rates, primary productivity on chlorophyll -a and cell basis, and maximum yield achieved in batch culture of three clones of Phaeodactylum tricornutum has been carried out in order to cheek intraespecifie differences among them. Two of the clones were isolated from samples of Uba-tuba (U3, isolated in 1972 and U5, in 1979 ), while the third one was isolated from samples of Cananéia (C1, isolated in 1980). Three-way analysis of variance was accom-plished and cluster analysis was carried out to distinguish the experimental conditions in which the clones would show distinct physiological responses. During the log phase of growth the three clones usually presented fusiform type of cells. Never theless, rare oval and triradiate cells have been observed. The pattern of the growth curves submitted todifferent temperatures (16,20,24º C) and media (Guillard f1. Erd-Schreiber, ASP2) was the same for the three clones. The results indieated significant differenees in growth rates and primary productivity. Oval morphotypes in clusters have been observed in all clones after the log phase under exausted media. The clone U5 has exhibited this be haviour in a prominent way. Many star-shape clusters, constituted of fusiform morphotypes linked by one of the extremities of each cell were observed in clone Cl. Un the other hand, triradiate and cruciform cells, and the formation of chains, were observed only in clone U3.

Através de bioensaios estáticos fo-ram comparadas as taxas máximas de crescimento, razão de fotossintese em termos de clorofila-a e de célula, bem como o rendimento final de tres clones de Phaeodacty-tum tricornutum visando verificar uma possível existência de diferenças intraespe-cificas entre eles. Dois desses clones foram isolados a partir de amostras de Uba-tuba (U3, isolado em 1972 e U5 isolado em 1979) e um de Cananéia (Cl, isolado em 1980). Todos os resultados foram submeti-dos a uma análise de variância trifatorial e a uma anállse de agrupamento com a finlidade de discriminar as condições experimentais nas quais os tres clones diferi-ram significativamente em suas respostas fisiológicas. Durante a fase exponencial de crescimento os tres clones se apresen-taram com morfótipos fusiformes raramente se encontrando morfótipos ovóides e trir-radiados. O padrão das curvas de cresci -mento dos tres clones em diferentes temperaturas (16, 20, 240 C) e meios de cultu-ra (ASP 2, Erd-Schreiber, Guillard fl) não diferiu. Os resultados obtidos indicaram a existência de diferenças significativas nas taxas de crescimento e produti-vidade primária. Morfótipo ovóides for-mando grumos foram observados nos tres clones após a fase exponencial e de modo conspícuo no clone U5. Nas mesmas condi-ções, o clone Cl apresentou uma grande quantidade de grumos, com a forma de uma estrela, constituidos por morfótipos fusiformes unidos convergentemente por uma das extremidades de cada célula. Por outro lado, morfótipos trirradiados e cru-ciformes bem como formação de cadeias, só foram observados no clone U3. / A comparison of the maximum growth rates, primary productivity on chlorophyll -a and cell basis, and maximum yield achieved in batch culture of three clones of Phaeodactylum tricornutum has been carried out in order to cheek intraespecifie differences among them. Two of the clones were isolated from samples of Uba-tuba (U3, isolated in 1972 and U5, in 1979 ), while the third one was isolated from samples of Cananéia (C1, isolated in 1980). Three-way analysis of variance was accom-plished and cluster analysis was carried out to distinguish the experimental conditions in which the clones would show distinct physiological responses. During the log phase of growth the three clones usually presented fusiform type of cells. Never theless, rare oval and triradiate cells have been observed. The pattern of the growth curves submitted todifferent temperatures (16,20,24º C) and media (Guillard f1. Erd-Schreiber, ASP2) was the same for the three clones. The results indieated significant differenees in growth rates and primary productivity. Oval morphotypes in clusters have been observed in all clones after the log phase under exausted media. The clone U5 has exhibited this be haviour in a prominent way. Many star-shape clusters, constituted of fusiform morphotypes linked by one of the extremities of each cell were observed in clone Cl. Un the other hand, triradiate and cruciform cells, and the formation of chains, were observed only in clone U3.

La méthylation de l'ADN est l’une des marques épigénétiques les plus étudiées et est largement conservée. Mes travaux de thèse présentent le premier méthylome d'une diatomée marine P. tricornutum qui appartient à la famille des Stramenopiles. P. tricornutum présente une méthylation d’environ 6% qui est présente en mosaïque sur l’ensemble du génome. Une méthylation importante a été retrouvé chez les éléments transposables, en particulier les éléments amplifiés récemment de type Copia. L’analyse met en évidence plus de 320 gènes méthylés dans trois contextes génomiques différents : à proximité des éléments transposables, en grappes de gènes méthylés, et dans des gènes uniques. En outre, les gènes largement et complètement méthylés ont été trouvé fortement corrélés avec le silencing transcriptionnel et l'expression différentielle dans des conditions spécifiques. Enfin, il a été constaté que les gènes susceptibles d’avoir été acquis par transfert horizontal de gènes bactériens étaient préférentiellement insérés dans des régions riches en éléments transposables, ce qui suggère un mécanisme par lequel l'expression de gènes étrangers peut être tamponnée à la suite de leur insertion dans le génome. En résumé, P. tricornutum a une faible méthylation de l'ADN et une methylation relativement importante des éléments transposables et seulement quelques gènes méthylés. Ce premier méthylome d’une diatomée Stramenopile ajoute de manière significative à notre compréhension de l'évolution de la méthylation de l'ADN chez les eucaryotes. En ce qui concerne les modifications des histones, la distribution des marques H3K4me2, H3K9me2 et H3K27me3 a été examinée chez P. tricornutum. H3K4me2 est principalement associée à des gènes alors que les deux marques H3K9me2 et H3K27me3 ciblent principalement des éléments transposables. La répartition de H3K27me3 est inhabituelle et différente de ce qui a été observé chez les espèces modèles étudiées à ce jour. Les gènes marqués par H3K27me3 ont tendance à être faiblement exprimés et de façon différentielle. H3K27me3 et H3K9me2 ont tendance à co-marquer non seulement les éléments transposables méthylés, mais aussi des gènes fortement méthylés, ce qui semble être important pour le maintien du silencing des gènes différentiellement exprimés. L'analyse combinatoire de différentes marques d’histones et la méthylation de l'ADN nous a donné un aperçu du paysage de la chromatine chez les diatomées, et aidera à définir les caractéristiques structurales et fonctionnelles conservées. / DNA methylation is the most extensively studied and widely conserved epigenetic mark. Here the first whole genome methylome from a stramenopile, the marine model diatom P. tricornutum is reported. In P. tricornutum, around 6% of the genome was methylated in a mosaic landscape. Extensive methylation in transposable elements (TEs), especially in recently amplified Copia-like elements was found. Over 320 genes were found methylated occurring in three different genomic contexts: in the proximity of TEs, in clusters of methylated genes, and in single genes. Furthermore, genes extensively and completely methylated correlated strongly with transcriptional silencing and differential expression under specific conditions. Finally, it was found that genes likely acquired by horizontal gene transfer from bacteria were preferentially inserted within TE-rich regions, suggesting a mechanism whereby the expression of foreign genes can be buffered following their insertion in the genome. In general, P. tricornutum has low DNA methylation with relatively extensive DNA methylation on TEs and a few methylated genes. This first Stramenopile methylome adds significantly to our understanding of the evolution of DNA methylation in eukaryotes. As for the histone modifications, genome wide distribution of H3K4me2, H3K9me2 and H3K27me3 were examined in P. tricornutum. H3K4me2 is mainly associated with genes while both H3K9me2 and H3K27me3 marks target mainly transposable elements (TEs). The distribution of H3K27me3 is unusual and different from what have been profiled in model species so far. The genes marked by H3K27me3 tend to be lowly and differentially expressed. H3K27me3 and H3K9me2 tend to co-mark not only methylated TEs but also heavily methylated genes, which appears to be important for maintaining the silencing of differentially expressed genes. The combinatorial analysis of different histone marks and DNA methylation gave us an overview of diatom chromatin landscapes, and will help to define conserved structural and functional features.

Depuis la découverte de Phaeodactylum tricornutum par Bohlin en 1897, sa classification au sein de l'arbre de la vie a été controversée. En utilisant des morphotypes ovales et fusiformes Lewin a décrit en 1958 plusieurs traits caractéristiques de cette espèce rappelant la structure des diatomées mettant ainsi fin à la controverse sur la classification de P. tricornutum au sein des Bacillariophycées. Pour se faire, trois morphotypes (ovale, fusiforme et triradié) de Phaeodactylum tricornutum ont été observés. Au cours d’une centaine d’années environ, de 1908 à 2000, 10 souches de Phaeodactylum tricornutum (appelées écotypes) ont été collectées et stockées soit de manière axénique ou en l’état avec leur populations naturelles de bactéries dans les centres des ressources génétiques pour algues, cryo-préservées quand cela est possible. Divers outils cellulaires et moléculaires ont été établis pour disséquer et comprendre la physiologie et l'évolution de P. tricornutum, et/ou les diatomées en général. Grâce à des décennies de recherche et les efforts déployés par de nombreux laboratoires que P. tricornutum est aujourd’hui considérée comme une espèce modèle des diatomées. Le sujet de ma thèse traite majoritairement de la composition génétique et épigénétique du génome de P. tricornutum ainsi que de la diversité morphologique et physiologique sousjacente au sein des populations naturelles prospectées à différents endroits du globe. Pour se faire, j’ai généré les profils chromatiniens en utilisant différentes marques des modifications post-traductionnelles des histones (chapitres 1 et 2) et a également comparé la variation naturelle dans la distribution de certaines marques clés entre deux populations d’écotypes (chapitre 4). Nous avons également généré une carte de la diversité génétique à l’échelle du génome chez 10 écotypes de P. tricornutum révélant ainsi la présence d'un complexe d'espèces dans le genre Phaeodactylum comme la conséquence d’une hybridation ancienne (chapitre 3). Sur la base de nombreux rapports antérieurs et des observations similaires au sein de P. tricornutum, nous proposons l’hybridation naturelle comme une base solide et une possibilité plausible pour expliquer la diversité des espèces chez lest diatomées. De plus, nous avons mis à jour les annotations fonctionnelles et structurelles du génome de P. tricornutum (Phatr3, chapitre 2) et mis au point un algorithme de logiciel convivial pour aller chercher les cibles CRISPR du système d’édition du génome CRISPR / cas9 chez 13 génomes de phytoplancton incluant P. tricornutum (chapitre 5). Pour accomplir tout cela, j'ai utilisé diverses méthodes à la pointe de l’état de l’art comme la spectrométrie de masse, l’immunoprécipitation de la chromatine suivie de séquençage à haut débit ainsi que les séquençages du génome entier, de l'ARN et des protocoles d'édition du génome CRISPR et plusieurs logiciels / pipelines de calcul. Ainsi, le travail de thèse fournit une plate-forme complète qui pourra être utilisée à l’avenir pour des études épigénétiques, de génétiques moléculaires et fonctionnelles chez les diatomées en utilisant comme espèce modèle Phaeodactylum tricornutum. Ce travail est pionnier et représente une valeur ajoutée importante dans le domaine de la recherche sur les diatomées en répondant à des questions nouvelles ouvrant ainsi de nouveaux horizons à la recherche en particulier en épigénétique qui joue un rôle important mais pas encore assez apprécié dans le succès écologique des diatomées dans les océans actuels. / Since the discovery of Phaeodactylum tricornutum by Bohlin in 1897, its classification within the tree of life has been controversial. It was in 1958 when Lewin, using oval and fusiform morphotypes, described multiple characteristic features of this species that resemble diatoms structure, the debate to whether classify P. tricornutum as a member of Bacillariophyceae was ended. To this point three morphotypes (oval, fusiform and triradiate) of Phaeodactylum tricornutum have been observed. Over the course of approximately 100 years, from 1908 till 2000, 10 strains of Phaeodactylum tricornutum (referred to asecotypes) have been collected and stored axenically as cryopreserved stocks at various stock centers. Various cellular and molecular tools have been established to dissect and understand the physiology and evolution of P. tricornutum, and/or diatoms in general. It is because of decades of research and efforts by many laboratories that now P. tricornutum is considered to be a model diatom species. My thesis majorly focuses in understanding the genetic and epigenetic makeup of P. tricornutum genome to decipher the underlying morphological and physiological diversity within different ecotype populations. To do so, I established the epigenetic landscape within P. tricornutum genome using various histone post-translational modification marks (chapter 1 and chapter 2) and also compared the natural variation in the distribution of some key histone PTMs between two ecotype populations (chapter 4). We also generated a genome-wide genetic diversity map across 10 ecotypes of P. tricornutum revealing the presence of a species-complex within the genus Phaeodactylum as aconsequence of ancient hybridization (Chapter 3). Based on the evidences from many previous reports and similar observations within P. tricornutum, we propose natural hybridization as a strong and potential foundation for explaining unprecedented species diversity within the diatom clade. Moreover, we updated the functional and structural annotations of P. tricornutum genome (Phatr3, chapter 2) and developed a user-friendly software algorithm to fetch CRISPR/Cas9 targets, which is a basis to perform knockout studies using CRISPR/Cas9 genome editing protocol, in 13 phytoplankton genomes including P. tricornutum (chapter 5). To accomplish all this, I used various state-of-the-art technologies like Mass-Spectrometry, ChIPsequencing, Whole genome sequencing, RNA sequencing, CRISPR genome editing protocols and several computational softwares/pipelines. In brief, the thesis work provides a comprehensive platform for future epigenetic, genetic and functional molecular studies in diatoms using Phaeodactylum tricornutum as a model. The work is an addon value to the current state of diatom research by answering questions that have never been asked before and opens a completely new horizon and demand of epigenetics research that underlie the ecological success of diatoms in modern-day ocean.

La protection intégrée, qui vise à réduire l’usage des pesticides, est un défi majeur pour l’agriculture du XXIème siècle. Le développement de nouvelles approches agronomiques qui concilient environnement et agriculture est une condition indispensable pour l’agriculture de demain. Dans ce contexte, l’utilisation d’éliciteurs capables de mimer une attaque pathogène et de promouvoir un état de résistance chez les plantes face à des maladies représente une alternative naturelle à la lutte chimique. Ces éliciteurs sont nommés les stimulateurs de défense des plantes (SDP). Ils peuvent provenir de différentes sources et être extraits à partir de macroalgues comme c’est le cas des SDP à base de polysaccharides d’algues tels que la laminarine utilisée pour stimuler l’immunité de plantes agronomiques. Toutefois, l’exploitation de ces ressources dans leur milieu naturel et les difficultés de production liées à leur cycle de développement constituent des freins à leur utilisation. La valorisation des microalgues comme source de SDP pourrait permettre de s’affranchir de ces contraintes. Cependant la recherche et de molécules SDP chez les microalgues est encore peu abordée. Au cours de ce travail, le potentiel d’une culture de microalgue, Phaeodactylum tricornutum, à induire des réactions de défense chez les plantes a été évalué. Un broyat cellulaire a été appliqué sur des plantules d’Arabidopsis thaliana. Le caractère éliciteur de ce broyat a été testé et caractérisé par des approches microscopiques, physiologiques et moléculaires. Les résultats ont montré que les plantes traitées présentaient des niveaux d’expression des gènes PR-1, PAD3, ACS6 et WRKY40 et un niveau de protection contre la bactérie Pseudomonas syringae DC3000 (Pst) plus élevés que les plantes non traitées. De plus, un effet bactéricide in vitro sur la bactérie Pst a été observé. Ces résultats offrent de nouvelles perspectives pour le développement de produits SDP d’origine naturelle capables de protéger les cultures. / Integrated plant protection, which aims to reduce the use of pesticide, is a major challenge for the agriculture of the 21st century. The development and application of new agronomic approaches is a prerequisite for crop protection in a sustainable agriculture system. In this context, the use of elicitors capable of mimicking a pathogenic attack and promoting a plant resistance state against diseases is a natural alternative to the use of agro-chemicals. These elicitors are also called plant defense stimulators (PDS). These can be obtained from different sources including macroalgae as it the case for the polysaccharide-based PDS laminarin that is currently used for the protection of a number of crops. However, the exploitation of these natural resources and the difficulties of their production due to their development cycle do hamper their use at a large scale. One of the possibilities to overcome these difficulties is the use of microalgae as a source of PDS. But this possibility and the potential of microalgaederived PDS for crop protection are currently under investigated. In the present work, we have used a cell extract from the microalgae Phaeodactylum tricornutum and assessed its defense response-eliciting activities on Arabidopsis thaliana seedlings by using microscopic, physiological and molecular approaches. The results show that treated plants exhibit higher levels of expression of the PR-1, PAD3, ACS6 and WRKY40 genes and a higher level of protection against the pathogenic bacterium Pseudomonas syringae DC3000 (Pst) than nontreated plants. An In vitro antibacterial activity on the Pst bacteria was also observed. Our findings suggest that P. tricornutum cell extracts are able to activate plant immune responses and offer new perspectives for the development of novel plant defense stimulators.

Actuellement, plus de 70% des biomédicaments commercialisés sont des glycoprotéines recombinantes. Les coûts élevés de production de ces biomédicaments ont poussé les scientifiques à développer des organismes de production alternatifs. Récemment, les microalgues ont été proposées en tant que potentiel système de production compte-tenu de leur rapidité de croissance et de leurs faibles coûts de production. Cependant, avant de produire des biomédicaments industriels chez les microalgues, il est impératif de s’assurer que les modifications post-traductionnelles, comme la N-glycosylation, soit conservées et compatibles avec une utilisation thérapeutique. Dans ce contexte, l’étude de la Nglycosylation de deux microalgues modèles, Chlamydomonas reinhardtii (microalgue verte) et Phaeodactylum tricornutum (diatomée) a été réalisée. Dans un premier temps, l’ingénierie de la N-glycosylation de C. reinhardtii a été initiée en exprimant une Nacétylglucosaminyltransférase I (GnT I) hétérologue. Les résultats obtenus ont permis de réévaluer les voies de N-glycosylation de C. reinhardtii et de montrer que cette microalgue synthétise une structure glycannique linéaire qui n’est pas substrat de la GnT I. Dans un second temps, un protocole d’extraction et de caractérisation des précurseurs glycanniques de C. reinhardtii et P. tricornutum a été développé et appliqué pour déterminer la structure des précurseurs glycanniques dans ces espèces. Enfin, la caractérisation de deuxxylosyltransférases potentielles (XTA et XTB) de C. reinhardtii a été menée en utilisant des mutants d’insertion et des analyses des N-glycannes par spectrométrie de masse. Cette étude a confirmé les rôles spécifiques de XTA et XTB dans la voie de N-glycosylation de C. reinhardtii. / Currently, more than 70% of the commercialized biopharmaceuticals are glycoproteins. The high production costs lead scientists to develop alternative organisms suitable for such production. Recently, microalgae emerged as a potential interesting production system thanks to their quick growth rate and low production costs. However, prior to start industrial glycoproteins production in microalgae, protein post-translational modifications like Nglycosylation, must be carefully controlled. This PhD thesis focused on the analysis of the Nglycosylation pathway of two different microalgae, Chlamydomonas reinhardtii (greenmicroalgae) and Phaeodactylum tricornutum (diatom). In order to start N-glycan engineering, heterologous N-acetylglucosaminyltransferase I (GnT I) sequences were expressed in C.reinhardtii. This study demonstrated that C. reinhardtii synthetize a linear N-glycan unsuitable for GnT I activity and allows the reinvestigation of the C. reinhardtii N-glycosylation pathway. A second chapter of this work focus on the optimization of a protocol suitable for analyzing the structure of the Dolichol N-linked precursors of C. reinhardtii and P. tricornutum. Lastly, two potential xylosyltransferases (XTA and XTB) from C. reinhardtii were characterized using insertional mutants and N-glycomic analyses by mass spectrometry approaches. This work allows us to propose specific involvement of XTA and XTB in the xylosylation processing of C.reinhardtii N-glycans.

This thesis reports the rheological properties of algae slurries as a function of cell concentration for three microalgae species: Nannochloris sp.,Chlorella vulgaris, and Phaeodactylum tricornutum. Rheological properties ofalgae slurries have a direct impact on the agitation and pumping power requirements as well as process design for producing algal biofuels. This study measures the rheological properties of eight diff erent concentrations of each species ranging from 0.5 to 80 kg dry biomass/m³. Strain-controlled steady rate sweep tests were performed for each sample with an ARES-TA rheometer using a double wall couette cup and bob attachment. Shear rates ranged from 5 - 270 s⁻¹, corresponding to typical expected conditions. The results showed that Nannochloris sp. slurry behaved as a Newtonian fluid for concentrations up to 20 kg/m³. Samples with concentrations above 40 kg/m³ behaved as a shear thinning non-Newtonian fluid. The effective viscosity increased with increased biomass concentration for a maximum value of 3.3x10⁻³ Pa-s. Similarly, C. vulgaris slurry behaved as a Newtonian fluid with concentrations of up to 40 kg/m³, above which it displayed a shear thinning non-Newtonianf behavior and a maximum eff ective viscosity of 3.5x10⁻² Pa-s. On the other hand, P. tricornutum slurry demonstrated solely Newtonian fluid behavior, with the dynamic viscosity increasing with increasing biomass concentration for a maximum value of 3.2x10⁻³ Pa-s. The maximum observed e ffective viscosity occurred at a concentration of 80 kg/m³ for all three species. Moreover, an energy analysis was performed where a non-dimensional bioenergy transport e ffectiveness was de termined as the ratio of the energy content of the transported algae biomass to the sum of the required pumping power and the harvesting power. The results show that the increase in major losses due to increase in viscosity was overcompensated by the increase in the transported biomass energy. Also, cultivating a more concentrated slurry requires less dewatering power and is the preferred option. The largest bioenergy transport eff ectiveness was observed for the slurries with the largest initial dry biomass concentrations. Finally, the relative viscosity of algae slurries was modeled using a Kelvin-Voit based model for dilute and concentrated viscoelastic par- ticle suspensions. The model, which depends primarily on the packing factor of the algae species, agrees with the measured viscosity with an average error of 18%, while the concentrated particle suspension model was slightly more accurate than the dilute suspension model. / text

Microalgae are gaining more attention for several reasons such as being potential producers of sustainable fuel, for use as health supplements and in skincare. Simris Alg is a Swedish company that produces Omega 3 supplements from a primary producer of these fatty acids - the algal diatom Phaeodactylum tricornutum, which is a sustainable alternative to Omega 3 derived from fish. Omega 3 fatty acids constitute a small fraction of the total algal biomass, and to increase profitability and utilize all of the biomass, the purpose of this thesis project is to present potential applications for the residual material that is left after oil extraction. A general composition study was made of Simris Alg algal residue material, and results are compared to those found in previous studies of P. tricornutum biomass. An optimization of the fractionation is needed to separate the storage carbohydrate chrysolaminarin and cell wall component glucuronomannan, followed by analysis for confirmation. Also, it would be interesting to separate chitin, if there is any, since despite the presence of chitin synthases, it is unclear whether the diatom actually produces chitin. When gathering information, no actual experimental characterization of carbohydrate active enzymes involved in synthesis of the main carbohydrates investigated were found. Such information would be useful to increase production of the carbohydrate of interest, if valuable applications are found. Potential applications of various cell components, such as carbohydrates, in skincare would be interesting to investigate, as well as optimizing fucoxanthin extraction for use as an additional high value product next to Omega 3.