Enhancement of acid resistance of Scenedesmus dimorphus by acid adaptation

When using flue gas as carbon source for microalgae cultivation, the resulting acidic environment caused by SO _X and NO _X can inhibit microalgal growth. In this study, Scenedesmus dimorphus acquired increased acid resistance by prior exposure to sublethal acid stress; a process defined as acid adaptation. Among the five algal species tested, S. dimorphus showed the highest level of acid tolerance to extreme acid challenge (exposure to pH 3.0). Non-adapted and acid-adapted exponential algal cells were used as inocula for tubular photobioreactors aerated with 2 % CO₂. Previously adapted at pH 4.0 for 1 h, S. dimorphus developed highest growth rate under extreme acidic condition, and the maximum biomass concentration and specific growth rate at pH 3.0 (3.638?±?0.074 g?L^?1 and 1.037?±?0.008 d^?1, respectively) were respectively 14.22 and 10.79 % higher than those of non-adapted cells. Moreover, acid-adapted cells could tolerate lower pH of 2.5, at which the growth of non-adapted cells was totally inhibited. All the results indicated that acid adaptation was an effective approach for the acid resistance enhancement of microalgae.     

Effects of different nitrogen, phosphorus, and iron concentrations and salinity on lipid production in newly isolated strain of the tropical green microalga, Scenedesmus dimorphus KMITL

The fatty acid composition, the effect of different concentrations of nitrogen (16.5-344 mg ?L^?1), phosphorus (9–45 mg? L^?1), iron (9–45 mg? L^?1) and salinity levels (0–20 psu) on lipid production in the green microalga Scenedesmus dimorphus KMITL, a new strain isolated from a tropical country, Thailand, were studied. The alga was isolated from a freshwater fish pond, and cultured in Chlorella medium by varying one parameter at a time. The main fatty acid composition of this strain was C16–C18 (97.52 %) fatty acids. A high lipid content was observed in conditions of 16.5 mg? L^?1-N, or 22 mg ?L^?1-P, or 45 mg ?L^?1-Fe, or 5 psu salinity, which accumulated lipids to 20.3?±?0.4, 19.4?±?0.2, 24.7?±?0.5, and 14.3?±?0.2 % of algal biomass, respectively. Increasing lipid content and lipid productivity was noted when the alga was cultured under high iron concentration and high salinity, as well as under reduced phosphorus conditions, whereas nitrogen limitation only resulted in an increased lipid content.     

Development of genetic transformation methodologies for an industrially-promising microalga: Scenedesmus almeriensis

The development of the microalgal industry requires advances in every aspect of microalgal biotechnology. In this regard, the availability of genetic engineering tools for industrially-promising species is key. As Scenedesmus almeriensis has promise for industrial use, we describe here an Agrobacterium-based methodology that allows stable genetic transformation of it for the first time, thus opening the way to its genetic manipulation. Transformation was accomplished using two different antibiotic resistance genes [hygromicine phophotransferase (hpt) and Shble] and it is credited by PCR amplification of both hpt/Shble and GUS genes and by the β-glucuronidase activity of transformed cells. Nevertheless, the single 35S promoter seems unable to direct gene expression to a convenient level in S. almeriensis as suggested by the low GUS enzymatic activity. Temperature was critical for the transformation efficiency.     

Effects of temperature and pH on growth and antioxidant content of the microalga Scenedesmus obliquus

Reactive forms of oxygen can damage DNA (among other molecules), thus triggering, e.g., atherogenesis and carcinogenesis. However, such dietary antioxidants as lutein and β-carotene can effectively inactivate them; these compounds were found to high levels in a novel strain (M2-1) of the microalga Scenedesmus obliquus. The independent and combined effects of pH and temperature on its rates of growth and production of antioxidants were experimentally assessed, via a full factorial experimental design; the effects of each parameter independently, and of their interactions were accordingly quantified by ANOVA. Our results indicated that temperature plays a more important role on the maximum specific growth rate than pH; in terms of antioxidant content, pH and, to a lesser extent, temperature also have relevant effects. Consequently, the highest rate of biomass specific growth (0.294 ± 0.013 day(-1)) and biomass productivity (0.837 ± 0.054 mg L(-1) day(-1)) were associated with relatively low pH (6) and relatively high temperature (30°C). Conversely, the antioxidant production rate increased with pH; hence, the highest productivity (0.638 mg L(-1) day(-1)) was attained at pH 8 and 30°C. At the best operating conditions for antioxidant content, the levels of lutein and β-carotene were 203.57 ± 1.41 and 18.20 ± 0.33 mg mL(-1), respectively; the maximum production of either one occurred at the early exponential phase.     

Growth and lipid accumulation properties of a freshwater microalga Scenedesmus sp. under different cultivation temperature

Microalgal lipid is a promising feedstock for biodiesel production. Effect of cultivation temperature on the growth and lipid accumulation properties of a freshwater microalga Scenedesmus sp. LX1 was studied. Scenedesmus sp. LX1 could grow in a wide range of temperature (10～30°C), and the growth activation energy was 49.3 kJ·mol(-1). The optimal temperature to produce microalgal biomass and lipid was 20°C, and after 15 days of batch cultivation the productivities of 313.3 g biomass·(g P)(-1), 112 g lipid (g P)(-1) and 14.7 g TAGs·(g P)(-1) were obtained. The content of polyunsaturated fatty acids decreased with the increase of cultivation temperature. The reactive oxygen species (ROS) levels at 10°C and 20°C were higher than that under higher temperature. For the first time the cultivation temperature, ROS level, specific growth rate and lipid content per microalgal biomass were correlated together.     

Cultivation of Scenedesmus dimorphus using anaerobic digestate as a nutrient medium

Bioprocess and Biosystems Engineering - In this study, the microalga Scenedesmus dimorphus was cultivated phototrophically using unsterilized anaerobic digestate as a nutrient medium. A bench-scale...     

Waste utilization and biodiesel production by the green microalga Scenedesmus obliquus

Scenedesmus obliquus was cultivated in three types of waste discharges to couple waste treatment with biodiesel production. The lipid pool accumulation was boosted to 1.0 g liter(-1) against 0.1 g liter(-1) for the control. The waste-grown S. obliquus showed an increase in the content of the saturated fatty acid pool, which is desirable for good-quality biodiesel.     

Heavy metals removal by the microalga Scenedesmus incrassatulus in continuous cultures

The microalga Scenedesmus incrassatulus was grown under continuous regime in the presence of chromium(VI), cadmium(II) and copper(II), as single metal species and as mixtures of two or three metals, in a laboratory scale system. We used an artificial wastewater with low free ion activities (as determined by MINEQL+) due to the presence of EDTA (a strong chelating agent) but with total concentrations not suitable for acceptable environments. Chromium(VI) and cadmium(II) had positive interaction that increased the removal percentages of both these metals; we could not, however, detect any interaction with copper(II). S. incrassatulus was able to remove all the tested metals to some extent (25-78%), but bivalent metals were not removed as efficiently as reported in batch cultures, probably due to the high pH values there recorded. Chromium(VI) was more efficiently removed in continuous cultures than in batch culture, because the uptake of chromate could be favored by actively growing algae.     

Regiospecific reduction of adrenosterone to 11-ketotestosterone by microalga T76 Scenedesmus quadricauda

Summary Adrenosterone is reduced to 11-ketotestosterone by cultures of microalga Scenedesmus quadricauda. The best bioconversion yields (62%) were obtained in cultures containing 3×10⁶ cells/mL of alga and 100 mg of adrenosterone in 400 mL of medium after 10 days at 24°C with a photoperiod light - dark 16:8 h.     

Biodiesel production by the green microalga Scenedesmus obliquus in a recirculatory aquaculture system

Biodiesel production was examined with Scenedesmus obliquus in a recirculatory aquaculture system with fish pond discharge and poultry litter to couple with waste treatment. Lipid productivity of 14,400 liter ha(-1) year(-1) was projected with 11 cultivation cycles per year. The fuel properties of the biodiesel produced adhered to Indian and international standards.     

Physiological responses of the green microalga Acutodesmus dimorphus to temperature induced oxidative stress conditions

Temperature is the most critical factor that directly affects the physiological functioning and metabolic activities of any organism. With rising global temperature, understanding the heat stress response of an organism is critically important. In the present study, we investigated differences in the early changes occurring upon heat stress in the green microalga Acutodesmus dimorphus, a potential strain for biofuel production. The cells were heat-stressed at 45 and 50°C for 24 h and the temporal response of cells in terms of growth, pigments content, levels of oxidative stress biomarkers i.e., reactive oxygen species (ROS) and the response of enzymatic and non-enzymatic antioxidant scavengers were evaluated. The results revealed that after 24 h of heat stress at 45°C, the accumulations of chlorophyll a and carotenoids remained stable; all three ROS increased with the higher activities of various enzymatic and non-enzymatic antioxidants. On the contrary, at a higher temperature of 50°C, the accumulations of chlorophyll a, carotenoids and non-enzymatic antioxidants reduced drastically while the accumulations of all three ROS and the response of enzymatic antioxidants were significantly higher than those at 45°C. These results suggest that the cells utilize several stress acclimatization mechanisms to cope up the heat stress. There was a dramatic difference in the physiological changes and cellular antioxidant mechanism upon heat stress at 45 and 50°C. The cellular defense response of A. dimorphus gets impaired after heat stress at 50°C but remains active at 45°C, exhibiting the heat resistance and, thus, the thermotolerance.     

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.     

Characterization,extraction and purification of lutein produced by an indigenous microalga Scenedesmus obliquus CNW-N

This study aimed to improve the commercial viability of microalgae-based lutein production using an isolated microalga Scenedesmus obliquus CNW-N possessing a high lutein content of over 0.25%. Effective lutein extraction protocols, appropriate storage methods, and purification procedures were developed. Disruption of microalgae cells was most efficient with a bead-beater. The conventional saponification step was modified to reduce the overall extraction time by 24 h. Diethyl ether exhibited the best lutein extraction efficiency. Storage of the lutein extract at low temperature (4 or −20 °C) with antioxidant addition (around 0.01% BHT) can maintain 90% lutein stability after 80 days. Addition of a suitable amount of the antioxidant could promote the stability of lutein extracts under the exposure of light. The protocol developed in this work allows efficient lutein extraction from S. obliquus CNW-N at a lower cost. Further purification was employed to elevate the purity of lutein and its commercial value.     

Use of the microalga Scenedesmus obliquus to remove cadmium cations from aqueous solutions

The ability of a wild strain of Scenedesmus obliquus, isolated from a heavy metal-contaminated environment, to remove Cd²⁺ from aqueous solutions was studied at several initial concentrations. Viable biomass removed metal to a maximum extent of 11.4 mg_Cd/g at 1 mg_Cd/l, with most Cd²⁺ being adsorbed onto the cell surface. A commercially available strain (ACOI 598) of the same microalga species was also exposed to the same Cd concentrations, and similar results were obtained for the maximum extent of metal removal. Heat-inactivated cells removed a maximum of 6.04 mg_Cd/g at 0.5 mg_Cd/l. The highest extent of metal removal, analyzed at various pH values, was 0.09 mg_Cd/g at pH 7.0. Both strains of the microalga tested have proven effective in removing a toxic heavy metal from aqueous solutions, hence supporting their choice for bioremediation strategies of industrial effluents.     

Effect of acetate on growth and ammonium uptake in the microalga Scenedesmus obliquus

The effect of acetate on growth and rate of ammonium uptake in Scenedesmus obliquus (UTEX 78) was investigated under light-limiting conditions. Addition of acetate to autotrophic cells with a growth constant of 0.71 day⁻¹ resulted in an increase in the growth rate (mixotrophy, k = 1.3 day⁻¹), and in the presence of acetate, growth occurred in the dark (heterophy, k = 0.44 day⁻¹). The rate of ammonium uptake in autotrophy (17.8 amol cell⁻¹ min⁻¹) was similar to that in heterotrophy (17.4 amol cell⁻¹ min⁻¹) but was 3.7 times lower than that in mixotrophy (65.9 amol cell⁻¹ min⁻¹). In general, mixotrophic cells showed optimum ammonium uptake at the acetate concentration at which they were grown. In autotrophy, uptake of ammonium leveled off at about 12.5 μ M while no saturation was observed in mixotrophic cells. An increase in the rate of uptake of ammonium was observed in autotrophic cells within 1 h after the addition of acetate. The activity of isocitrate lyase (EC 4.1.3.1), a key enzyme for the regulation of the glyoxylate cycle responsible for acetate catabolism, showed a 3.9-fold increase in activity after 24 h in the dark in the presence of acetate. The level of isocitrate lyase activity in cells grown for 24 h in the dark in the presence of 0–20 m M acetate also increased as a function of acetate concentration.     

Ultrahigh-cell-density heterotrophic cultivation of the unicellular green microalga Scenedesmus acuminatus and application of the cells to photoautotrophic culture enhance biomass and lipid production

Although production of biodiesels from microalgae is proved to be technically feasible, a commercially viable system has yet to emerge. High-cell-density fermentation of microalgae can be coupled with photoautotrophic cultivation to produce oils. In this study, by optimizing culturing conditions and employing a sophisticated substrate feed control strategy, ultrahigh-cell-density of 286 and 283.5 g/L was achieved for the unicellular alga Scenedesmus acuminatus grown in 7.5-L bench-scale and 1,000-L pilot-scale fermenters, respectively. The outdoor scale-up experiments indicated that heterotrophically grown S. acuminatus cells are more productive in terms of both biomass and lipid accumulation when they are inoculated in photobioreactors for lipid production as compared to the cells originally grown under photoautotrophic conditions. Technoeconomic analysis based on the pilot-scale data indicated that the cost of heterotrophic cultivation of microalgae for biomass production is comparable with that of the open-pond system and much lower than that of tubular PBR, if the biomass yield was higher than 200 g/L. This study demonstrated the economic viability of heterotrophic cultivation on large-scale microalgal inocula production, but ultrahigh-productivity fermentation is a prerequisite. Moreover, the advantages of the combined heterotrophic and photoautotrophic cultivation of microalgae for biofuels production were also verified in the pilot-scale.     

A series of mutant strains of Scenedesmus obliquus with abnormal carotenoid compositions

Several mutant strains of Scenedesmus obliquus (Chlorophyta) have been isolated which, when cultured heterotrophically, are pale green or yellow, in contrast to the dark green of the wild type. On the basis of their carotenoid compositions, four groups of pale-green strains have been delineated. These accumulate, respectively, no carotenoid, phytoene, mainly -carotene and mainly -carotene together with some neurosporene and lycopene. All these strains synthesized no chlorophyll b and only small amounts of chlorophyll a. A further group of yellow strains produced the normal Scenedesmus obliquus range of cyclic carotenes and xanthophylls, but no chlorophyll. Most of the pale-green strains were killed by exposure to light, but two strains, PG1 and 1E, which accumulated predominantly -carotene when grown in the dark, survived exposure to the light and developed photosynthetically active chloroplasts containing the normal pigments.The possible biosynthetic implications of the carotenoid composition of these mutant strains, and the relationship between the carotenoid composition and protection of the cells from photooxidative destruction are discussed.Non-Standard Abbreviations TLC thin-layer chromatography