Morpho-physiological aspects of Scenedesmus acutus PVUW12 cultivated with a dairy industry waste and after starvation

Among green microalgae, Scenedesmus sp. is known for its potential in wastewater remediation and lipid production, especially under starvation. Moreover, it is often characterised by a mixotrophic metabolism. In this work, we cultivated S. acutus PVUW12 in the presence of a liquid fraction of scotta (LFS), a cheese whey by-product, as source of nutrients. Subsequently, cultures were starved to evaluate lipid production. Cells were analysed to obtain information about growth, nutrient consumption during LFS cultivation, morphology and photosynthetic efficiency. We found that the alga boosted its growth when cultured in presence of LFS. Production of stromatic starch grains, polyphosphate granules, cell wall enlargement and reduction of the photosynthetic efficiency were also induced. Massive lipid accumulation was observed only during starvation, which also induced a strong slowdown of growth, loss of polyphosphate grains and further decrease in photosynthetic efficiency. This study demonstrates that S. acutus PVUW12 can be involved in a two-step cultivation, first by promoting growth using a by-product from cheese industry and second by transferring the microalgae on starvation to induce lipid accumulation for bioenergetics purposes.     

Biomass production and fatty acid profile of a Scenedesmus rubescens-like microalga

This investigation examined the effects of nitrogen–phosphate combined deficiency on the biomass yield, fatty acid methyl esters (FAME) production and composition from Scenedesmus rubescens-like microalga. A 15-day indoor culture was performed as a 3 × 3 factorial design (NaNO₃ levels: 3, 10 and 20 mM; KH₂PO₄ levels: 20, 50 and 150 μM). The algae grown under medium nitrogen concentration (10 mM) and high phosphate concentration (150 μM) reached the highest biomass (1223.5 ± 152.5 mg/L). Both nitrogen and phosphate had a significant influence on the FAME yield (P < 0.05 and P < 0.0001, respectively). The FAME yield from algae grown under low nitrogen (3 mM) and phosphate concentration (20 μM) increased throughout the experiment and the highest FAME yield (42.2 ± 2.5% of AFDW) as well as C16 and C18 content (95.8 ± 1.6% of AFDW) was achieved under these conditions. Algae grown under medium nitrogen concentration (10 mM) and low phosphate concentration (20 μM) had the highest FAME productivity (426.0 mg/L ± 135.0 mg/L). Thus, the lower nitrogen concentration (3 mM–10 mM) and low phosphate concentration (20 μM) would be an optimal combination tested to produce the most FAME from S. rubescens-like algae.     

Studies on Scenedesmus acutus growth II. Effect of autotrophic and mixotrophic growth on the amino acid and the carbohydrate composition of Scenedesmus acutus

As a general rule an increase in carbohydrates occurs during the light phase of the cell cycle and that of protein during phase, although variations were found in these components under autotrophic and mixotrophic growth conditions. The results are based on the quantitative determination of carvohydrates as trimethylsilyl (TMS) derivatives and amino acids as N-trifluoroacetyl-n-butyl (TAB) esters in algal cells cultured in light and dark periods by gas-liquid chromatography (LC). Cells harvested during the dark period contained more amino acids as compared to similar cultures harvested during the light phase. In light, the production of amino acids of the aspartate family increased in cells cultivated with glucose and carbon dioxide. With glucose as sole carbon source, the carbohydrate content was higher in the dark than in the light period. Under continuous light conditions, in the presence of carbon dioxide, there was a decrease in the carbohydrate content also. Gas-liquid chromatography analysis of the extract of the purified cell walls showed that they are made up of 0.076% carbohydrates and 0.28% amino acids on the dry weight (DW) basis of whole cells. The results on the metabolism of cells, under autotrophic and mixotrophic conditions, are discussed in this article.     

Studies on Scenedesmus acutus growth. The incorporation of 14C-glucose by Scenedesmus acutus under light and dark periods

Qualitative and quantitative estimations of alcohol-extractable compounds from (14)C-glucose-incorporated Scendesmus acutus cells were performed at various intervals of time. After two hours of incubation with (14)C-glucose in light, amino acids were synthesized at 65% and sucrose at 26% of the total input label. In the dark incorporation, 30% of the total radioactivity was found in amino acids and 46% in sucrose. Leucine and valine were detected only during the oxidation of glucose in light. The concentration of serine increased more in the presence of light, as compared to dark. These results on the oxidation of glucose are discussed in this article.     

Optimization of biomass and fatty acid productivity of Scenedesmus obliquus as a promising microalga for biodiesel production

Nowadays, microalgae are discussed as a promising feedstock for biodiesel production. The present study examines the possibility of enhancement of fatty acid productivity of Scenedesmus obliquus by modifications of the culture medium composition. The effect of different concentrations of sodium bicarbonate, salinity, potassium nitrate, glycerol and sugarcane molasses on the enhancement of biomass and esterified fatty acids production was studied. NaHCO₃ caused an increase in the biomass productivity at low concentrations (0.5 g L^?1), while negatively affected fatty acid productivity at all tested concentrations. Increase of salinity enhanced both biomass and fatty acid productivity. The optimum NaCl concentration and sea water ratio were 0.94 g L^?1 and 25 % which resulted in 56 and 39 % increase in fatty acid productivity, respectively. Nitrogen deficiency showed increase in fatty acid content by 54 % over control but fatty acid productivity was decreased as a result of growth inhibition. Nitrogen-free cultures and cultures treated with ?50 % concentrations of KNO₃ showed 96 and 42 % decrease in EFA productivity, respectively, as compared with the control. Addition of 0.05 and 0.1 M of glycerol increased the biomass productivity by 6 and 5 %, respectively but showed no significant effect on fatty acid productivity as a result of decrease in fatty acid content. Finally, usage of sugarcane molasses stimulated both biomass and fatty acid content. The increase in fatty acid productivity was 32, 65 and 73 % above the control level at 1, 3 and 5 g L^?1 of sugarcane molasses, respectively.     

Studies on Scenedesmus acutus growth. I effect of autotrophic and mixotrophic conditions on the growth of Scenedesmus acutus

Among sugars, glucose and mannose were found to be the most suitable substrates for mixotrophic growth, uptake of galactose and its influence on growth was negligible, and sucrose and fructose occupied intermediary positions. The optimum temperature for sugar uptake was 30 degrees C, both under light and in darkness. Enhancement in the photosynthetic oxygen-evolution rate, based on the utilization of substrates, was foremost in the presence of glucose, followed by mannose, sucrose, and fructose. Industrial by-products such as sugarcane molasses also were utilized to increase the algal growth under mixotrophic conditions. A maximum yield in biomass was obtained subsequent to the combined supply of sugarcane molasses with carbon dioxide to indoor as well as outdoor mixotrophic cultures. Doubling the carbon dioxide supply alone above a certain level, under autotrophic and mixotrophic outdoor conditions, did not produce a pronounced increase in the algal growth rate. The results on autotrophic and mixotrophic growth variations are discussed in the article.     

Biomass, lipid content, and fatty acid composition of freshwater Chlamydomonas mexicana and Scenedesmus obliquus grown under salt stress

Two freshwater microalgae including Chlamydomonas mexicana and Scenedesmus obliquus were grown on Bold Basal Medium (BBM) with different levels of salinity up to 100 mM NaCl. The dry biomass and lipid content of microalgae were improved as the concentration of NaCl increased from 0 to 25 mM. Highest dry weight (0.8 and 0.65 g/L) and lipid content (37 and 34 %) of C. mexicana and S. obliquus, respectively, were obtained in BBM amended with 25 mM NaCl. The fatty acid composition of the investigated species was also improved by the increased NaCl concentration. At 50 mM, NaCl palmitic acid (35 %) and linoleic acid (41 %) were the dominant fatty acids in C. mexicana, while oleic acid (41 %) and α-linolenic acid (20 %) were the major fractions found in S. obliquus.     

Endurance in high-fat-fed rats: effects of carbohydrate content and fatty acid profile

The purpose ofthis experiment was to study endurance performance and substratestorage and utilization in fat- or carbohydrate-fed rats. Ninety-ninerats were randomly divided into three groups and over 4 wk were fedeither a carbohydrate-rich [CHO; 10% total energy content in the diet(E%) fat, 20 E% protein, 70 E% carbohydrate] diet or one of twofat-rich diets (65 E% fat, 20 E% protein, 15 E% carbohydrate)containing either saturated (Sat) or monounsaturated fatty acids(Mono). Each dietary group was randomly assigned to a trained (6 days/wk, progressive to 60 min, 28 m/min at a 10% incline) or asedentary group. Rats were killed either before or after a treadmillendurance run to exhaustion. Training increased endurance (206%), butdiet composition did not affect endurance in either trained orsedentary rats. -Hydroxyacyl-CoA dehydrogenase activity wasincreased in fat-fed but not carbohydrate-fed rats (P < 0.05). Respiratory exchangeratio during the initial phase of exercise was lower after the Monocompared with the Sat diet (P < 0.05) and higher after the CHO than the Sat diet(P < 0.05). Thus adaptation to ahigh-fat diet containing a moderate amount of carbohydrates did notinduce enhanced endurance in either trained or untrained rats; however,substrate utilization was modulated by both amount and type of dietaryfat during the initial stage of exercise in trained and sedentary rats.

     

Production technology of Scenedesmus acutus in the Indian context

Summary The West German Dortmund production procedure for the green alga Scenedesmus acutus has been practically evaluated under Indian conditions utilising 179 m² of cultivable area to produce about 26.7 kg of dried algae per week. For a 16 day out-door production schedule, cleaning of cultivation tanks takes 27% of the total man hours required, and indoors inoculum preparation takes 20%. Electrical energy consumption depends mainly on the drying mode used but irrespective of any change in the system the cost structure remains high for Indian contexts.     

Electron-dense Regions in Dividing Mitochondria of a Green Alga, Scenedesmus acutus

The dividing and the constricted regions of mitochondria ina green alga, Scenedesmus acutus , were studied by electronmicroscopy. Electron-dense substances were always visible inthese specific regions of mitochondria that had been fixed byfreeze-substitution. Thin fibres, which seemed to surround theconstricted regions, were also seen. The possibility that theelectron-dense regions constitute part of the dividing apparatusof the mitochondria is discussed.Copyright 1993, 1999 AcademicPress Division apparatus, division of mitochondria, electron microscopy, mitochondria, Scenedesmus     

Triacylglycerol fatty acid and sterol composition of sediment microorganisms from McMurdo Sound,Antarctica

Summary The triacylglycerol fatty acid and sterol profiles of microorganisms from three McMurdo Sound sediment sites, collected during the austral summer of 1984–1985, were determined using gas chromatography and gas chromatography-mass spectrometry. Comparison of the three sites indicated that Cape Evans contained the greatest concentration of triacylglycerol (TG) (220 nmoles/gram dry weight (gdw) of sediment), approximately six to seven times that determined for sediment microorganisms from the Cape Armitage and New Harbor sites. The relative proportion of triacylglycerolderived polyunsaturated fatty acids (PUFA) revealed a somewhat different trend. New Harbor sediment contained the greatest relative proportion of PUFA (22% of triacylglycerol fatty acids), followed by Cape Evans (16%) and Cape Armitage (11%). The proportion of unsaturated fatty acids (poly-and monounsaturated) was relatively constant and ranged from 63% to 71% of the triacylglycerol fatty acids for the three sites. Sterol concentrations varied from 610 pmoles/gdw at Cape Evans, to 370 and 240 pmoles/gdw for Cape Armitage and New Harbor respectively, and was approximately 1% of the total determined lipid. Cholesterol was the major sterol component detected, occurring at similar relative levels (29%) for all three sites. Other sterols present in decreasing order of abundance were 22-dehydrocholesterol, brassicasterol, 24-ethylcholesterol and 24-methylcholesterol. 5-stanols were only minor components of the three sediments, indicating that in situ biohydrogenation of stenols was not a major sterol transformation process in these recent surface oxic sediments.Part 3 in the series: Microbial Ecology in Antarctic Sea-Ice and Benthic Communities     

Bioenergetic analysis of mixotrophic growth in Chlorella vulgaris and Scenedesmus acutus

The specific growth rate of Chlorella vulgaris in the presence of glucose (mixotroph) was always larger than that in autotroph when the light intensity was less than 10 klux. However, Scenedesmus acutus behaved differently, i.e., when the light intensity was more than 6 klux, the specific growth rate in mixotroph became smaller than that in autotroph. Cellular contents of chlorophyll a and b in Scenedesmus acutus that deteriorated more markedly in mixotroph than those of Chlorella vulgaris could account for these different growth behaviors. In other words, mechanisms relevant to the photosynthesis and the oxidative phosphorylation of glucose seem to function independently with respect to Chlorella Vulugaris.     

Growth,lipid content,productivity, and fatty acid composition of tropical microalgae for scale‐up production

Biomass and lipid productivity, lipid content, and quantitative and qualitative lipid composition are critical parameters in selecting microalgal species for commercial scale‐up production. This study compares lipid content and composition, and lipid and biomass productivity during logarithmic, late logarithmic, and stationary phase of Nannochloropsis sp., Isochrysis sp., Tetraselmis sp., and Rhodomonas sp. grown in L1‐, f/2‐, and K‐medium. Of the tested species, Tetraselmis sp. exhibited a lipid productivity of 3.9–4.8 g m^?2 day^?1 in any media type, with comparable lipid productivity by Nannochloropsis sp. and Isochrysis sp. when grown in L1‐medium. The dry biomass productivity of Tetraselmis sp. (33.1–45.0 g m^?2 day^?1) exceeded that of the other species by a factor 2–10. Of the organisms studied, Tetraselmis sp. had the best dry biomass and/or lipid production profile in large‐scale cultures. The present study provides a practical benchmark, which allows comparison of microalgal production systems with different footprints, as well as terrestrial systems. Biotechnol. Bioeng. 2010;107: 245–257. © 2010 Wiley Periodicals, Inc.     

两步法培养对丰富栅藻脂肪酸和氨基酸的影响

为了提高丰富栅藻的油脂产量和评估培养条件对其营养成分的影响,对丰富栅藻的培养基正交优化后,对其进行两步法培养并分析培养前后的脂肪酸和氨基酸的变化。结果表明：两步法培养后丰富栅藻的生物量、油脂含量和油脂产量分别达到23.57 g/L、33.49%和7.58 g/L; 丰富栅藻的单不饱和脂肪酸（MUFAs）特别是C18：1（n-9）的相对含量有所上升,但是饱和脂肪酸（SFAs）和多不饱和脂肪酸（PUFAs）的相对含量有所下降; 氨基酸特别是必需氨基酸的总量降低; 但丝氨酸和蛋氨酸的含量却略有增加。     

Lipid productivity, settling potential and fatty acid profile of 11 microalgal species grown under nitrogen replete and limited conditions

Microalgae are a promising alternative source of oil for biodiesel production. Identification of a species with desirable characteristics is a key component towards achieving economic feasibility for the process. This has been compromised by a lack of data allowing effective interspecies comparison. Eleven species of microalgae, selected on the basis of available literature data, were tested for lipid productivity, gravity sedimentation and the suitability of their fatty acid profiles for biodiesel production. The response to nitrogen limitation was species-specific. Lipid yields and productivity were higher at 150?mg?L^?1 nitrate than at 1,500?mg?L^?1 for all species tested except Spirulina platensis. The Chlorophyta, particularly Chlorella vulgaris and Scenedesmus, had the highest growth rates and showed the greatest increase in lipid content in response to nitrogen limitation. Cylindrotheca fusiformis, S. platensis, Scenedesmus and Tetraselmis suecica had the fastest settling rates and highest biomass recoveries after 24?h of gravity sedimentation. For most species, the fuel would need to be blended or culture conditions to be optimised to achieve the correct lipid profile in order for microalgal fuel to meet the European standards for biodiesel production (EN 14214). The most promising species overall were the freshwater algae Scenedesmus and C. vulgaris and the marine algae C. fusiformis and Nannochloropsis.     

Influence of light on plastocyanin formation in the alga Scenedesmus acutus

Scenedesmus acutus, pregrown autotrophically, forms high levels of plastocyanin during an initiation period of 24 hr in the light, after cupric ions have been added to depleted cells. In the dark, no plastocyanin formation is observed. This is in contrast to heterotrophic cells which, over the same period and under identical conditions, yield about half the plastocyanin level regardless of whether they are incubated in the dark or in the light. The ATP level is high in all cases when substantial formation of plastocyanin takes place, while the ³⁵S-sulfolipid level, the marker of thylakoid formation, is at variance. After the 24-hr plastocyanin formation period, the ³⁵S-sulfolipid level is high in illuminated autotrophic cells as well as in dark-incubated heterotrophic ones, but low in illuminated heterotrophic cells, all of them having substantial plastocyanin contents. Additional experiments on light dependence of copper uptake, influence of light quality, occurrence of apoplastocyanins, provide a set of data to assume that the role of light in plastocyanin formation is primarily through photophosphorylation, the ATP presumably being necessary for processing of plastocyanin precursors to holoprotein.     

Triacylglycerol content of arachidonic acid varies inversely with total triacylglycerol in liver and plasma

In both liver and plasma from rats, hamsters, guinea pigs and monkeys, the correlation coefficients between percentage composition of arachidonic acid and the total triacylglycerol varied from -0.43 to -0.81 (P less than 0.01). No such relationship was present between percentage compositions of any of the essential fatty acids in phospholipids and total phospholipids.     

Effects of eicosapentaenoic acid on synaptic plasticity,fatty acid profile and phosphoinositide 3-kinase signaling in rat hippocampus and differentiated PC12 cells

Placebo-controlled clinical studies suggest that intake of n-3 polyunsaturated fatty acids improves neurological disorders such as Alzheimer's disease, Huntington's disease and schizophrenia. To evaluate the impact of eicosapentaenoic acid (EPA), we orally administered highly purified ethyl EPA (EPA-E) to rats at a dose of 1.0 mg/g per day and measured long-term potentiation of the CA1 hippocampal region, a physiological correlate of synaptic plasticity that is thought to underlie learning and memory. The mean field excitatory postsynaptic potential slope of the EPA-E group was significantly greater than that of the control group in the CA1 region. Gene expression of hippocampal p85α, one of the regulatory subunits of phosphatidylinositol 3-kinase (PI3-kinase), was increased with EPA-E administration. Investigation of fatty acid profiles of neuronal and glia-enriched fractions demonstrated that a single administration of EPA-E significantly increased neuronal and glial EPA content and glial docosahexaenoic acid content, clearly suggesting that EPA was indeed taken up by both neurons and glial cells. In addition, we investigated the direct effects of EPA on the PI3-kinase/Akt pathway in differentiated PC12 cells. Phosphorylated-Akt expression was significantly increased in EPA-treated cells, and nerve growth factor withdrawal-induced increases in cell death and caspase-3 activity were suppressed by EPA treatment. These findings suggest that EPA protects against neurodegeneration by modulating synaptic plasticity and activating the PI3-kinase/Akt pathway, possibly by its own functional effects in neurons and glial cells and by its capacity to increase brain docosahexaenoic acid.