The Effect of Trophic Modes on Biomass and Lipid Production of Five Microalgal Strains

Five microalgae strains, namely Isochrysis galbana, Microchloropsis gaditana, Scenedesmus obliquus, Nannochloropsis oculata and Tetraselmis suecica, were selected as potential candidates for polyunsaturated fatty acids’ production, evaluating biomass productivity and their capacity to accumulate high lipid contents under different trophic modes. Microalgae strains were cultivated in the presence of 1% glucose using mixotrophic and heterotrophic conditions, while autotrophic cultures served as control experiments. The results demonstrate that S. obliquus performed the highest biomass productivity that reached 0.13 and 0.14 g L−1 d−1 under mixotrophic and heterotrophic conditions, respectively. I. galbana and S. obliquus utilized elevated contents of glucose in mixotrophy, removing 55.9% and 95.6% of the initial concentration of the carbohydrate, respectively, while glucose consumption by the aforementioned strains also remained high under heterotrophic cultivation. The production of lipids was maximal for I. galbana in mixotrophy and S. obliquus in heterotrophy, performing lipid productivities of 24.85 and 22.77 mg L−1 d−1, respectively. The most abundant saturated acid detected for all microalgae strains evaluated was palmitic acid (C16:0), while oleic and linolenic acids (C18:1n9c/C18:3n3) comprised the most abundant unsaturated fatty acids. I. galbana performed the highest linoleic acid (C18:2n6c) content under heterotrophic nutrition, which reached 87.9 mg g−1 of ash-free dry weight. Among the microalgae strains compared, the biomass and lipid production monitored for I. galbana and S. obliquus confirm that both strains could serve as efficient bioproducers for application in algal biorefineries.

Palm Oil Mill Effluent for Lipid Production by the Diatom Thalassiosira pseudonana

Biomass and lipid production by the marine centric diatom Thalassiosira pseudonana were characterized in media based on palm oil mill effluent (POME) as a source of key nutrients. The optimal medium comprised 20% by volume POME, 80 µM Na2SiO3, and 35 g NaCl L−1 in water at pH ~7.7. In 15-day batch cultures (16:8 h/h light–dark cycle; 200 µmol photons m−2 s−1, 26 ± 1 °C) bubbled continuously with air mixed with CO2 (2.5% by vol), the peak concentration of dry biomass was 869 ± 14 mg L−1 corresponding to a productivity of ~58 mg L−1 day−1. The neutral lipid content of the biomass was 46.2 ± 1.1% by dry weight. The main components of the esterified lipids were palmitoleic acid methyl ester (31.6% w/w) and myristic acid methyl ester (16.8% w/w). The final biomass concentration and the lipid content were affected by the light–dark cycle. Continuous (24 h light) illumination at the above-specified irradiance reduced biomass productivity to ~54 mg L−1 day−1 and lipid content to 38.1%.

The Effect of Nutrients Mixture on The Biomass and Lipid Production from Microalgae Botryococcus braunii Mutated by UV-C Rays

Nutrient is one of the most important factors in the growth of microalgae. This research was conducted to study the effect of nutrient mixture on the biomass and lipid production of Botryococcus braunii. Microalgae B. braunii was cultivated in the commercial nutrient medium of agricultural fertilizer combinations of ammonium sulphate (ZA), urea, and triple superphosphate (TSP). Before the cultivation process, B. braunii was exposed to UV-C rays (254 nm) for 3 minutes. The concentration and type of fertilizer as a nitrogen source divided into four types of mixtures, namely FM-1, FM-2, FM-3, and FM-4 were compared with Walne nutrients to study their effects on microalgae growth and lipids. FM-1 consisting of 150 mg/L of ZA, 7.5 mg/L of urea, and 25 mg/L of TSP led to the best growth for native and mutated microalgae strains compared to Walne nutrients and other nutrient mixtures. The mutated microalgae showed less growth than the native microalgae strains. However, the mutation process significantly increased the lipid content in the microalgae. In native microalgae strains, FM-4 consisting of 136.3 mg/L of urea and 50 mg/L of TSP produced the lowest lipid at 8.96%. After being exposed to UV-C rays, the lipids in FM-4 medium increased to 55.11%. The results show that the use of commercial fertilizers and exposure to UV-C rays on microalgae have high potential in preparing lipids as raw material for biodiesel which can be effectively applied in large-scale microalgae cultivation.

Highly Valuable Polyunsaturated Fatty Acids from Microalgae: Strategies to Improve Their Yields and Their Potential Exploitation in Aquaculture

Microalgae have a great potential for the production of healthy food and feed supplements. Their ability to convert carbon into high-value compounds and to be cultured in large scale without interfering with crop cultivation makes these photosynthetic microorganisms promising for the sustainable production of lipids. In particular, microalgae represent an alternative source of polyunsaturated fatty acids (PUFAs), whose consumption is related to various health benefits for humans and animals. In recent years, several strategies to improve PUFAs’ production in microalgae have been investigated. Such strategies include selecting the best performing species and strains and the optimization of culturing conditions, with special emphasis on the different cultivation systems and the effect of different abiotic factors on PUFAs’ accumulation in microalgae. Moreover, developments and results obtained through the most modern genetic and metabolic engineering techniques are described, focusing on the strategies that lead to an increased lipid production or an altered PUFAs’ profile. Additionally, we provide an overview of biotechnological applications of PUFAs derived from microalgae as safe and sustainable organisms, such as aquafeed and food ingredients, and of the main techniques (and their related issues) for PUFAs’ extraction and purification from microalgal biomass.

Triacyl Glycerols from Yeast-Catalyzed Batch and Fed-Batch Bioconversion of Hydrolyzed Lignocellulose from Cardoon Stalks

The lipogenic ability of the yeast Solicoccozyma terricola DBVPG 5870 grown on hydrolyzed lignocellulose obtained from cardoon stalks was evaluated. Data on cell biomass, lipid production, and fatty acid profiles of triacylglycerols obtained in batch and fed-batch experiments were carried out at the laboratory scale in a 5L fermenter, and at two different temperatures (20 and 25 °C) were reported. The higher production of total intracellular lipids (13.81 g/L) was found in the fed-batch experiments carried out at 20 °C. S. terricola exhibited the ability to produce high amounts of triacylglycerol (TAGs) with a characteristic fatty acids profile close to that of palm oil. The TAGs obtained from S. terricola grown on pre-treated lignocellulose could be proposed as a supplementary source of oleochemicals. Indeed, due to the rising prices of fossil fuels and because of the environmental-related issues linked to their employment, the use of TAGs produced by S. terricola grown on lignocellulose could represent a promising option as a supplementary oleochemical, especially for biodiesel production.

Tricarboxylate Citrate Transporter of an Oleaginous Fungus Mucor circinelloides WJ11: From Function to Structure and Role in Lipid Production

The citrate transporter protein (CTP) plays an important role in citrate efflux from the mitochondrial matrix to cytosol that has great importance in oleaginous fungi. The cytoplasmic citrate produced after citrate efflux serves as the primary carbon source for the triacylglycerol and cholesterol biosynthetic pathways. Because of the CTP's importance, our laboratory has extensively studied its structure/function relationships in Mucor circinelloides to comprehend its molecular mechanism. In the present study, the tricarboxylate citrate transporter (Tct) of M. circinelloides WJ11 has been cloned, overexpressed, purified, kinetically, and structurally characterized. The Tct protein of WJ11 was expressed in Escherichia coli, isolated, and functionally reconstituted in a liposomal system for kinetic studies. Our results showed that Tct has a high affinity for citrate with Km 0.018 mM. Furthermore, the tct overexpression and knockout plasmids were created and transformed into M. circinelloides WJ11. The mitochondria of the tct-overexpressing transformant of M. circinelloides WJ11 showed a 49% increase in citrate efflux, whereas the mitochondria of the tct-knockout transformant showed a 39% decrease in citrate efflux compared to the mitochondria of wild-type WJ11. To elucidate the structure-function relationship of this biologically important transporter a 3D model of the mitochondrial Tct protein was constructed using homology modeling. The overall structure of the protein is V-shaped and its 3D structure is dimeric. The transport stability of the structure was also assessed by molecular dynamics simulation studies. The activity domain was identified to form hydrogen bond and stacking interaction with citrate and malate upon docking. Tricarboxylate citrate transporter has shown high binding energy of −4.87 kcal/mol to citric acid, while −3.80 kcal/mol to malic acid. This is the first report of unraveling the structural characteristics of WJ11 mitochondrial Tct protein and understanding the approach of the transporting toward its substrate. In conclusion, the present findings support our efforts to combine functional and structural data to better understand the Tct of M. circinelloides at the molecular level and its role in lipid accumulation.