Optimization of lipase production from yeasts strains isolated from olive mil wastewater

Dissertação submetida à Universidade de Lisboa, Faculdade de Ciências para a obtenção do Grau de Mestre em Microbiologia Aplicada.

Phytoplankton versus macrophyte contribution to primary production and biogeochemical cycles of a coastal mesotidal system. A modelling approach

This study presents an assessment of the contributions of various primary producers to the global annual production and N/P cycles of a coastal system, namely the Arcachon Bay, by means of a numerical model. This 3D model fully couples hydrodynamic with ecological processes and simulates nitrogen, silicon and phosphorus cycles as well as phytoplankton, macroalgae and seagrasses. Total annual production rates for the different components were calculated for different years (2005, 2007 and 2009) during a time period of drastic reduction in seagrass beds since 2005. The total demand of nitrogen and phosphorus was also calculated and discussed with regards to the riverine inputs. Moreover, this study presents the first estimation of particulate organic carbon export to the adjacent open ocean. The calculated annual net production for the Arcachon Bay (except microphytobenthos, not included in the model) ranges between 22,850 and 35,300 tons of carbon. The main producers are seagrasses in all the years considered with a contribution ranging from 56% to 81% of global production. According to our model, the -30% reduction in seagrass bed surface between 2005 and 2007, led to an approximate 55% reduction in seagrass production, while during the same period of time, macroalgae and phytoplankton enhanced their productions by about +83% and +46% respectively. Nonetheless, the phytoplankton production remains about eightfold higher than the macroalgae production. Our results also highlight the importance of remineralisation inside the Bay, since riverine inputs only fulfill at maximum 73% nitrogen and 13% phosphorus demands during the years 2005, 2007 and 2009. Calculated advection allowed a rough estimate of the organic matter export: about 10% of the total production in the bay was exported, originating mainly from the seagrass compartment, since most of the labile organic matter was remineralised inside the bay.

Effect of Inorganic and Organic Carbon Enrichments (DIC and DOC) on the Photosynthesis and Calcification Rates of Two Calcifying Green Algae from a Caribbean Reef Lagoon

Coral reefs worldwide are affected by increasing dissolved inorganic carbon (DIC) and organic carbon (DOC) concentrations due to ocean acidification (OA) and coastal eutrophication. These two stressors can occur simultaneously, particularly in near-shore reef environments with increasing anthropogenic pressure. However, experimental studies on how elevated DIC and DOC interact are scarce and fundamental to understanding potential synergistic effects and foreseeing future changes in coral reef function. Using an open mesocosm experiment, the present study investigated the impact of elevated DIC (pHNBS: 8.2 and 7.8; pCO2: 377 and 1076 μatm) and DOC (added as 833 μmol L-1 of glucose) on calcification and photosynthesis rates of two common calcifying green algae, Halimeda incrassata and Udotea flabellum, in a shallow reef environment. Our results revealed that under elevated DIC, algal photosynthesis decreased similarly for both species, but calcification was more affected in H. incrassata, which also showed carbonate dissolution rates. Elevated DOC reduced photosynthesis and calcification rates in H. incrassata, while in U. flabellum photosynthesis was unaffected and thalus calcification was severely impaired. The combined treatment showed an antagonistic effect of elevated DIC and DOC on the photosynthesis and calcification rates of H. incrassata, and an additive effect in U. flabellum. We conclude that the dominant sand dweller H. incrassata is more negatively affected by both DIC and DOC enrichments, but that their impact could be mitigated when they occur simultaneously. In contrast, U. flabellum can be less affected in coastal eutrophic waters by elevated DIC, but its contribution to reef carbonate sediment production could be further reduced. Accordingly, while the capacity of environmental eutrophication to exacerbate the impact of OA on algal-derived carbonate sand production seems to be species-specific, significant reductions can be expected under future OA scenarios, with important consequences for beach erosion and coastal sediment dynamics.

Potential of Trichoderma harzianum for control of banana leaf fungal pathogens when applied with a food source and an organic adjuvant

Trichoderma isolates were obtained from diseased leaves and fruit collected from plantations in the main banana production area in Northern Queensland. Phylogenetic analyses identified the Trichoderma isolates as T. harzianum and T. virens. The Trichoderma spp. were found to be antagonistic against the banana leaf pathogens Mycosphaerella musicola, Cordana musae, and Deight-oniella torulosa in vitro. Several products used by the banana industry to increase production, including molasses, Fishoil and Seasol, were tested as food source for the Trichoderma isolates. The optimal food substrate was found to be molasses at a concentration of 5 %, which when used in combination with a di-1-p-menthene spreader-sticker enhanced the survivability of Trichoderma populations under natural conditions. This formulation suppressed D. torulosa development under glasshouse conditions. Furthermore, high sensitivity was observed towards the protectant fungicide Mancozeb but Biopest oil (R), a paraffinic oil, only marginally suppressed the growth of Trichoderma isolates in vitro. Thus, this protocol represents a potential to manage banana leaf pathogens as a part of an integrated disease approach.

Production responses of reproducing ewes to a by-product-based diet inoculated with the probiotic Bacillus amyloliquefaciens strain H57

The potential application of the spore-forming probiotic Bacillus amyloliquefaciens strain H57 (H57) as a novel probiotic for ruminants was evaluated in reproducing ewes. Performance responses were determined by delivering H57 in a pelleted diet based mainly on palm kernel meal (PKM) and sorghum grain. PKM is an agro-industrial by-product with a reputation for poor palatability and the availability of the starch in sorghum grain can be limited in ruminants. The hypothesis was that H57 improves the feeding value of a relatively low quality concentrate diet. Twenty-four first-parity white Dorper ewes were fed PKM-based pellets manufactured with or without H57 (109 cfu/kg pellet) in late pregnancy. During this phase of late pregnancy, the H57 ewes ate 17% more dry matter (1019 vs 874 g/day, P = 0.03), gained more weight (194 vs 30 g/day, P = 0.008) and retained more nitrogen (6.13 vs 3.34 g/day, P = 0.01), but produced lambs with a similar birthweight (4.1 vs 4.2 kg, P = 0.73). Rumen fluid collected from H57 ewes in late pregnancy had higher pH (7.1 vs 6.8, P = 0.07), acetate : propionate ratio (3.4 vs 2.7, P = 0.04), lower ammonia (69 vs 147 mmol/L, P = 0.001) and total volatile fatty acid concentrations (40 vs 61 mg/L, P = 0.02). The digestibility of dry matter, organic matter and fibre were similar between the two groups. The lambs of the H57 ewes grew faster than those of the Control ewes for the first 21 days of lactation (349 vs 272 g/day, P = 0.03), but not thereafter. H57 can improve feed intake and maternal liveweight gain in late pregnancy of first-parity ewes fed a diet based on PKM.

Effects of organic fertilizers on the vegetative, nutritional, and productive parameters of blueberries ‘Corona’, ‘Legacy’, and ‘Liberty’

Organic farming does not allow using certain inputs, such as N, which differ in nutrient release rates and dynamics. To evaluate the effect of different organic fertilizers on the vegetative, nutritional, and productive parameters of blueberries ( Vaccinium corymbosum L.), a pot experiment was conducted in three consecutive seasons in a sandy soil of south-central Chile using ‘Corona’, ‘Legacy’ and ‘Liberty’. The following fertilizers were evaluated: compost (CM), Purely Grow (PG), Purely Lysine (PL), Fertil (F), blood meal (BM), lupine meal (LM), along with a control treatment without fertilization (C) and two conventional treatments with urea (CF) and sodium nitrate (S). Results indicate that vegetative growth and leaf N concentration prior to senescence were different among cultivars in the three evaluated seasons. The highest leaf N concentration was recorded in ‘Corona’ followed by ‘Legacy’ and ‘Liberty’ while levels tended to increase in the seasons. Quick-release N sources had greater effects on these parameters but with differences among cultivars. Fruit yield and weight were higher in ‘Corona’ followed by ‘Legacy’ and ‘Liberty’. Fruit yield was generally higher when using LM and F and showed no effect on fruit weight. Leaf chlorophyll content was higher in ‘Corona’ followed by ‘Legacy’ and ‘Liberty’, which increased when using CF, LM, BM, and PG. Finally, the organic fertilizer and blueberry cultivar that obtained the highest values for most of the evaluated parameters were LM and Corona, respectively.

Improving photofermentative hydrogen production through metabolic engineering and DOE (Design of Experiments)

A l’heure actuelle, les biocarburants renouvelables et qui ne nuit pas à l'environnement sont à l'étude intensive en raison de l'augmentation des problèmes de santé et de la diminution des combustibles fossiles. H2 est l'un des candidats les plus prometteurs en raison de ses caractéristiques uniques, telles que la densité d'énergie élevée et la génération faible ou inexistante de polluants. Une façon attrayante pour produire la H2 est par les bactéries photosynthétiques qui peuvent capter l'énergie lumineuse pour actionner la production H2 avec leur système de nitrogénase. L'objectif principal de cette étude était d'améliorer le rendement de H2 des bactéries photosynthétiques pourpres non sulfureuses utilisant une combinaison de génie métabolique et le plan des expériences. Une hypothèse est que le rendement en H2 pourrait être améliorée par la redirection de flux de cycle du Calvin-Benson-Bassham envers du système de nitrogénase qui catalyse la réduction des protons en H2. Ainsi, un PRK, phosphoribulose kinase, mutant « knock-out » de Rhodobacter capsulatus JP91 a été créé. L’analyse de la croissance sur des différentes sources de carbone a montré que ce mutant ne peut croître qu’avec l’acétate, sans toutefois produire d' H2. Un mutant spontané, YL1, a été récupéré qui a retenu l'cbbP (codant pour PRK) mutation d'origine, mais qui avait acquis la capacité de se développer sur le glucose et produire H2. Une étude de la production H2 sous différents niveaux d'éclairage a montré que le rendement d’YL1 était de 20-40% supérieure à la souche type sauvage JP91. Cependant, il n'y avait pas d'amélioration notable du taux de production de H2. Une étude cinétique a montré que la croissance et la production d'hydrogène sont fortement liées avec des électrons à partir du glucose principalement dirigés vers la production de H2 et la formation de la biomasse. Sous des intensités lumineuses faibles à intermédiaires, la production d'acides organiques est importante, ce qui suggère une nouvelle amélioration additionnel du rendement H2 pourrait être possible grâce à l'optimisation des processus. Dans une série d'expériences associées, un autre mutant spontané, YL2, qui a un phénotype similaire à YL1, a été testé pour la croissance dans un milieu contenant de l'ammonium. Les résultats ont montré que YL2 ne peut croître que avec de l'acétate comme source de carbone, encore une fois, sans produire de H2. Une incubation prolongée dans les milieux qui ne supportent pas la croissance de YL2 a permis l'isolement de deux mutants spontanés secondaires intéressants, YL3 et YL4. L'analyse par empreint du pied Western a montré que les deux souches ont, dans une gamme de concentrations d'ammonium, l'expression constitutive de la nitrogénase. Les génomes d’YL2, YL3 et YL4 ont été séquencés afin de trouver les mutations responsables de ce phénomène. Fait intéressant, les mutations de nifA1 et nifA2 ont été trouvés dans les deux YL3 et YL4. Il est probable qu'un changement conformationnel de NifA modifie l'interaction protéine-protéine entre NifA et PII protéines (telles que GlnB ou GlnK), lui permettant d'échapper à la régulation par l'ammonium, et donc d'être capable d'activer la transcription de la nitrogénase en présence d'ammonium. On ignore comment le nitrogénase synthétisé est capable de maintenir son activité parce qu’en théorie, il devrait également être soumis à une régulation post-traductionnelle par ammonium. Une autre preuve pourrait être obtenue par l'étude du transcriptome d’YL3 et YL4. Une première étude sur la production d’ H2 par YL3 et YL4 ont montré qu'ils sont capables d’une beaucoup plus grande production d'hydrogène que JP91 en milieu d'ammonium, qui ouvre la porte pour les études futures avec ces souches en utilisant des déchets contenant de l'ammonium en tant que substrats. Enfin, le reformage biologique de l'éthanol à H2 avec la bactérie photosynthétique, Rhodopseudomonas palustris CGA009 a été examiné. La production d'éthanol avec fermentation utilisant des ressources renouvelables microbiennes a été traitée comme une technique mature. Cependant, la plupart des études du reformage de l'éthanol à H2 se sont concentrés sur le reformage chimique à la vapeur, ce qui nécessite généralement une haute charge énergetique et résultats dans les émissions de gaz toxiques. Ainsi le reformage biologique de l'éthanol à H2 avec des bactéries photosynthétiques, qui peuvent capturer la lumière pour répondre aux besoins énergétiques de cette réaction, semble d’être plus prometteuse. Une étude précédente a démontré la production d'hydrogène à partir d'éthanol, toutefois, le rendement ou la durée de cette réaction n'a pas été examiné. Une analyse RSM (méthode de surface de réponse) a été réalisée dans laquelle les concentrations de trois facteurs principaux, l'intensité lumineuse, de l'éthanol et du glutamate ont été variés. Nos résultats ont montré que près de 2 moles de H2 peuvent être obtenus à partir d'une mole d'éthanol, 33% de ce qui est théoriquement possible.

Non chemical natural organic matter removal with a contact TiO2 process

Dissertação de Mestrado, Tecnologia dos Alimentos, Instituto Superior de Engenharia, Universidade do Algarve, 2014

A survey on the composition of wines made with grapes produced by an organic system.

Summary There is concern about the health problems caused by pesticides in humans, which has led some grape producers to adopt organic procedures in their vineyards, and a certain amount of these grapes are directed to winemaking. Despite the approval awarded to this organic grape production by the certified organizations, there has been a demand to carry out a survey to determine the physicochemical composition of the wine derived from these products. Some of these wines were made from a single grape variety and others from more than one. For this survey, the samples consisted of five bottles of each type of wine, acquired from wineries and supermarkets in the Serra Gaúcha region, RS, Brazil. The analyses were carried out by physicochemical methods: volatile compounds by gas chromatography; minerals and trace elements by inductively coupled plasma optical emission spectrometry; and pesticide residues by liquid chromatography-mass spectrometry. The results showed that in general the physicochemical composition of these wines was within the limits established by Brazilian legislation. The mineral and trace element concentrations were very low and pesticide residues were not detected (MRL = 10 μg.kg?1) in any of the wines. Resumo: Há preocupação em relação a problemas causados por pesticidas no ser humano, o que levou uma parcela de viticultores a adotar procedimentos de agricultura orgânica em seus vinhedos. Assim, devido a essa preocupação, eles estão produzindo uva pelo sistema orgânico, sendo que uma parte dessa produção é direcionada à elaboração de vinho. Apesar de essas uvas terem sido aprovadas por entidades certificadoras, houve demanda para a realização de um levantamento para determinar a composição dos vinhos delas derivados. Portanto, cinco garrafas de cada tipo de vinho foram coletadas em vinícolas e supermercados da Serra Gaúcha, Rio Grande do Sul, sendo uma parte de vinhos varietais e outra, de cortes de diferentes variedades. As análises foram feitas por métodos físico-químicos: compostos voláteis, por cromatografia gasosa; minerais e elementos-traço, por espectrometria de emissão ótica com plasma acoplado indutivamente, e resíduos de pesticidas, por cromatografia líquida-espectrometria de massa. Os resultados mostram que a composição físico-química desses vinhos e dos minerais situou-se, em geral, dentro dos limites da legislação brasileira. As concentrações de minerais e de elementos-traço foram muito baixas, e não foram detectados resíduos de pesticidas (LMR = 10 μg.kg?1) em nenhum vinho.

Potential for agricultural production on disturbed soils mined for apatite using legumes and beneficial microbe

Christmas Island has been mined for rock phosphate for over 100 years, and as mining will finish in the next few decades there is a need to develop alternative economies on the island, such as high value crop production. However, to conserve the unique flora and fauna on the island, only land previously mined will be considered for this purpose. As these soils have been severely perturbed by mining, strategies to improve soil quality parameters need to be undertaken before plant based industries can be considered. For instance, legumes and beneficial microbes have demonstrated a positive role in the remediation of degraded soils. Therefore, this study aimed to establish the scientific basis upon which agriculture can effectively be developed on s oils post phosphate mining. Six legume species (Glycine max (Soybean), Vigna radiata (Mungbean), V. unguiculata (Cowpea), Phaseolus vulgaris (Navybean), Cajanus cajan (Pigeon pea), and Lablab purpureus (Lablab)) were sown onto a two ha rehabilitated site t hat had previously been mined for rock phosphate. The soil had a pH of 7.0, and was high in P but low in Bo, Cu, K, Mg, N and S and had low organic C. The legumes were inoculated with their respective rhizobial inoculant or co-inoculated with the rhizobia and a plant growth promoting bacteria (PGPB) at three different fertilizer rates (nil, a low rate, and five times the low rate). With the exception of P. vulgaris, all the legume species survived. The application of fertilizer was essential for maximum biomass yields 18 weeks after sowing, however the lower fertilizer rate was sufficient to obtain maximum yields for some cultivars. The PGPB increased yields and nodulation of some of the legumes at different fertilizer levels. Although the legumes (except P. vulgaris) grew in the Christmas Island environment, selection of appropriate legume cultivars and inoculants plus optimization of the fertilizer regime is required for reliable agricultural productivity on the island.

The production and potential of biofertilizers to improve crop yields

Extensive interactions of plant roots with soil microorganisms affect plant nutrition either directly by influencing mineral nutrient availability or indirectly through enhanced uptake efficiency via plant root growth promotion. Beneficial microbial interactions with roots may be either endophytic or associative and can be symbiotic, mutualistic, or incidental in nature. The increased understanding of the role of root – or rhizosphere –associated with microbes in the nutrition and/or yield of agricultural crops in particular has resulted in promotion of their use in agricultural production as alternatives or supplements to mineral or organic fertilizers. Despite this, there is an obvious lack of market penetration of microbial inoculants. This review specifically focuses on microbial inoculants, collectively termed biofertilizers, used to improve nutrition and yields of grain, legume, oil, tuber, and other crops. A vast number of commercial biofertilizers are available worldwide; however, the quality and efficacy of many of them are not proven or tested. In the absence of efficacious biofertilizers of good and consistent quality, the dependence on the use of mineral fertilizers is not likely to decrease. Thus the availability of high-quality biofertilizers must be priority particularly in countries where crop plant production plays a key role in the economy and food security.

Biomethane production from macroalgae

Irish brown seaweeds have been identified as a potential bio-resource with potentially high specific methane yields. Anaerobic digestion is deemed the most feasible technology due to its commercial viability for handling such wet feedstock. However, the biomethane potential of seaweed is highly dependent on its chemical composition which can vary by species type, cultivation method, and time of harvest. This study aims to investigate and optimize the process for the production of biomethane from Irish brown seaweeds focusing on the key technology bottlenecks including for seaweed characterization, biomethane potential assessment, optimization of long-term anaerobic digestion and suitable pre-treatment technologies to enhance potential gas yields. Laminaria digitata and Ascophyllum nodosum were tested for seasonal variation. From the characterization and batch digestion of L. digitata, August was found to be the optimal month for harvest due to high organic matter content, low level of ash and ultimately highest biomethane yield. The specific methane yield of 53 m3 CH4 t-1 wwt in August was 4.5 times higher than the yield in December (12 m3 CH4 t-1 wwt), with ash content the key factor in seasonal variation. For A. nodosum, the optimal harvest month was October with polyphenol content found to be a more influential factor than ash. The gross energy yields from both species were evaluated in the range of 116-200 GJ ha-1 yr-1. Continuous digestion trials were subsequently designed for S. latissima and L. digitata to optimize the key digestion parameters. Results from mono-digestion and co-digestion with dairy slurry revealed that both seaweeds could be digested at maximum biomethane efficiency to a loading rate of 4 kg VS m-3 d-1. Accumulation of salt in the digesters was a concern for long term digestion and it was reasoned that suitable pretreatment may be required prior to digestion. Various pre-treatments were subsequently tested on L. digitata to enhance the gas yield. It was found that maceration after hot water washing yielded 25% more specific methane and up to 54% salt removal as compared to untreated L. digitata. The experiments undertaken aim to assist in providing a basic guideline for feasible design and operation of seaweed digesters in Ireland.

Debating food production at dinner time: Mainstream audiences are invited to debate the ethics of agricultural practices in Brazil

In the rural areas of Brazil, a farmer runs his agricultural empire with a fierce hand: he exploits his workers and the land to their limits. Lack of sustainable land management leads to the pollution of rivers, changes in rain patterns, and exhaustion of the soil.

Potential mitigation of midwest grass-finished beef production emissions with soil carbon sequestration in the United States of America

Beef production can be environmentally detrimental due in large part to associated enteric methane (CH4) production, which contributes to climate change. However, beef production in well-managed grazing systems can aid in soil carbon sequestration (SCS), which is often ignored when assessing beef production impacts on climate change. To estimate the carbon footprint and climate change mitigation potential of upper Midwest grass-finished beef production systems, we conducted a partial life cycle assessment (LCA) comparing two grazing management strategies: 1) a non-irrigated, lightly-stocked (1.0 AU/ha), high-density (100,000 kg LW/ha) system (MOB) and 2) an irrigated, heavily-stocked (2.5 AU/ha), low-density (30,000 kg LW/ha) system (IRG). In each system, April-born steers were weaned in November, winter-backgrounded for 6 months and grazed until their endpoint the following November, with average slaughter age of 19 months and a 295 kg hot carcass weight. As the basis for the LCA, we used two years of data from Lake City Research Center, Lake City, MI. We included greenhouse gas (GHG) emissions associated with enteric CH4, soil N2O and CH4 fluxes, alfalfa and mineral supplementation, and farm energy use. We also generated results from the LCA using the enteric emissions equations of the Intergovernmental Panel on Climate Change (IPCC). We evaluated a range of potential rates of soil carbon (C) loss or gain of up to 3 Mg C ha-1 yr-1. Enteric CH4 had the largest impact on total emissions, but this varied by grazing system. Enteric CH4 composed 62 and 66% of emissions for IRG and MOB, respectively, on a land basis. Both MOB and IRG were net GHG sources when SCS was not considered. Our partial LCA indicated that when SCS potential was included, each grazing strategy could be an overall sink. Sensitivity analyses indicated that soil in the MOB and IRG systems would need to sequester 1 and 2 Mg C ha-1 yr-1 for a net zero GHG footprint, respectively. IPCC model estimates for enteric CH4 were similar to field estimates for the MOB system, but were higher for the IRG system, suggesting that 0.62 Mg C ha-1 yr-1 greater SCS would be needed to offset the animal emissions in this case.

Production de biodiesel à partir de microalgues par catalyses homogène et hétérogène

Résumé : Au Canada, près de 80% des émissions totales, soit 692 Mt eq. CO[indice inférieur 2], des gaz à effet de serre (GES) sont produits par les émissions de dioxyde de carbone (CO[indice inférieur 2]) provenant de l’utilisation de matières fossiles non renouvelables. Après la Conférence des Nations Unies sur les changements climatiques, COP21 (Paris, France), plusieurs pays ont pour objectif de réduire leurs émissions de GES. Dans cette optique, les microalgues pourraient être utilisées pour capter le CO[indice inférieur 2] industriel et le transformer en biomasse composée principalement de lipides, de glucides et de protéines. De plus, la culture des microalgues n’utilise pas de terre arable contrairement à plusieurs plantes oléagineuses destinées à la production de biocarburants. Bien que les microalgues puissent être transformées en plusieurs biocarburants tels le bioéthanol (notamment par fermentation des glucides) ou le biométhane (par digestion anaérobie), la transformation des lipides en biodiesel pourrait permettre de réduire la consommation de diesel produit à partir de pétrole. Cependant, les coûts reliés à la production de biodiesel à partir de microalgues demeurent élevés pour une commercialisation à court terme en partie parce que les microalgues sont cultivées en phase aqueuse contrairement à plusieurs plantes oléagineuses, ce qui augmente le coût de récolte de la biomasse et de l’extraction des lipides. Malgré le fait que plusieurs techniques de récupération des lipides des microalgues n’utilisant pas de solvant organique sont mentionnées dans la littérature scientifique, la plupart des méthodes testées en laboratoire utilisent généralement des solvants organiques. Les lipides extraits peuvent être transestérifiés en biodiesel en présence d’un alcool tel que le méthanol et d’un catalyseur (catalyses homogène ou hétérogène). Pour la commercialisation du biodiesel à partir de microalgues, le respect des normes ASTM en vigueur est un point essentiel. Lors des essais en laboratoire, il a été démontré que l’extraction des lipides en phase aqueuse était possible afin d’obtenir un rendement maximal en lipides de 36% (m/m, base sèche) en utilisant un prétraitement consistant en une ébullition de la phase aqueuse contenant les microalgues et une extraction par des solvants organiques. Pour l’estérification, en utilisant une résine échangeuse de cations (Amberlyst-15), une conversion des acides gras libres de 84% a été obtenue à partir des lipides de la microalgue Chlorella protothecoïdes dans les conditions suivantes : température : 120°C, pression autogène, temps de réaction : 60 min, ratio méthanol/lipides: 0.57 mL/g et 2.5% (m/m) Amberlyst-15 par rapport aux lipides. En utilisant ces conditions avec une catalyse homogène (acide sulfurique) et une seconde étape alcaline avec de l’hydroxyde de potassium (température : 60°C ; temps de réaction : 22.2 min; ratio catalyseur microalgue : 2.48% (m/m); ratio méthanol par rapport aux lipides des microalgues : 31.4%), un rendement en esters méthyliques d’acides gras (EMAG) de 33% (g EMAG/g lipides) a été obtenu à partir des lipides de la microalgue Scenedesmus Obliquus. Les résultats démontrent que du biodiesel peut être produit à partir de microalgues. Cependant, basé sur les présents résultats, il sera necessaire de mener d’autre recherche pour prouver que les microalgues sont une matière première d’avenir pour la production de biodiesel.