Effects of phosphorus concentration and light intensity on the biomass composition of Arthrospira (Spirulina) platensis

This paper presents the effects of various phosphorus concentrations (10, 50, 250 and 500 mg l(-1) K(2)HPO(4)) on the biomass production and composition of Arthrospira (Spirulina) platensis in relation to light intensity (24, 42 and 60 μE m(-2) s(-1)). The maximum biomass production was 3,592 ± 392 mg l(-1) and this was observed in 250 mg l(-1) K(2)HPO(4) at 60 μE m(-2) s(-1) light intensity after 32 days of cultivation. A maximum specific growth rate (μ(max)) of 0.55 d(-1) was obtained in 500 mg l(-1) K(2)HPO(4) at 60 μE m(-2) s(-1). The protein, lipid and chlorophyll contents of the biomass varied from 33.59 to 60.57 %, 5.34 to 13.33 % and 0.78 to 2.00 %, respectively. The most significant finding was that phosphorus limitation (10 mg l(-1) K(2)HPO(4)) caused a drastic increase of the carbohydrate content (59.64 %). The effect of phosphorus limitation on the carbohydrate content was independent of the light intensity. The accumulated carbohydrates are proposed to be used as substrate for biofuel generation via one of the appropriate biomass energy conversion technologies. Also, it was observed that phosphorus removal is a function of biomass density, phosphorus concentration and light intensity.     

Influence of ammonium chloride feeding time and light intensity on the cultivation of Spirulina (Arthrospira) platensis

This study dealt with the influence of both the feeding time and light intensity on the fed-batch culture of the cyanobacterium Spirulina (Arthrospira) platensis using ammonium chloride as a nitrogen source. For this purpose, a 2(2) plus star central composite experimental design combined with response surface methodology was employed, and the maximum cell concentration (X(m)), the cell productivity (P(X)), and the yield of biomass on nitrogen (Y(X/N)) were selected as the response variables. The optimum values of X(m) (1,833 mg L(-1)) and Y(X/N) (5.9 g g(-1)) estimated by the model at light intensity of 13 klux and feeding time of 17.2 days were very close to those obtained experimentally under these conditions (X(m) = 1,771 +/- 41 mg L(-1); Y(X/N) = 5.7 +/- 0.17 g g(-1)). The cell productivity was a decreasing function of the ammonium chloride feeding time and a quadratic function of the light intensity. The protein and lipid contents of dry biomass collected at the end of cultivations were shown to decrease with increasing light intensity.     

Arthrospira (Spirulina) platensis: An effective biosorbent for nutrients

Arthrospira (Spirulina) platensis was tested for biosorption properties. Preliminary experiments concerning biosorption kinetics were performed on Cr(III) ions. Equilibrium of biosorption was tested for Cr(III), Mn(II) and Mg(II) ions, since these elements are crucial for animals with metabolic disorders. In our study, Spirulina was proposed as a feed additive for animals suffering from diseases characterized by insulin dysregulation, abnormal adipose distribution and a high risk for laminitis. Maximum biosorption capacity of A. platensis, determined from Langmuir equation, was 45.2 for Cr(III), 44.3 for Mn(II) and 42.0 mg/g for Mg(II) ions. Biosorption of Mg(II) ions by microalga has never been studied so far. Finally, the raw and enriched microalgal biomass was examined by ICP-OES to determine its multielamental analysis before and after biosorption, FTIR to indicate functional groups that participated in biosorption and SEM-EDX to illustrate the binding of metal ions on the surface of algal biomass. ICP-OES showed that the content of elements significantly increased in the enriched A. platensis. FTIR spectroscopy evidenced that biosorption of metal ions was mainly due to carboxylate groups present on the microalgal cell wall. SEM analysis clearly showed that biosorption occurred. Arthrospira platensis turned out to be a good biosorbent of metal ions.     

First description of a cyanophage infecting the cyanobacterium Arthrospira platensis (Spirulina)

Cyanobacteria constitute a versatile group of photosynthetic bacteria of immense commercial and ecological importance. Some species of this group are cultivated and sold as food because of their high nutritional value. This is typically the case for Arthrospira platensis. We describe, for the first time, a virus infecting this economically important filamentous cyanobacterium isolated from culture pools located in the South of France. This virus could be observed and discriminated easily from other particles with flow cytometry. Based on morphology and molecular investigation, it was proposed that the virus belongs to the cyanopodovirus group with a capsid and short tail of about 120 and 20 nm, respectively. Finally, the virus appeared to be highly specific (very narrow host range) to A. platensis.     

Factors indicating culture status during cultivation of Spirulina (Arthrospira) platensis

Factors indicating culture status of two Spirulina platensis strains were monitored in a batch mode cultivation for 36 days. Changing mode in all factors showed a common turning point, indicating shift of cell or culture status. Mean biomass productivity was highly sustained until day 22, chlorophyll a concentration peaked on day 22, pH value was >12 on day 22, coil number was abruptly shortened on day 22, and floating activity was sustained at greater than 79% after day 22, indicating that day 22 is a criterion reflecting phase-transfer in cell physiology in a batch culture system. Many of these changes may have been caused by increased pH, suggesting that pH control is essential for mass production of S. platensis. Fluctuations in floating activity were likely induced by the number of cellular gas vacuoles. Consequently, coil number per trichome and floating activity of S. platensis could readily act as simple indicators for determination of culture status or harvesting time of cells.     

Photosynthetic and respiratory electron transport in the alkaliphilic cyanobacterium Arthrospira (Spirulina) platensis

Photosynthetic and respiratory electron transport and their interplay with ion transport have been studied in Arthrospira platensis, a filamentous alkaliphilic cyanobacterium living in hypersaline lakes. As typical for alkaliphiles, A. platensis apparently does not maintain an outward positive pH gradient at its plasma membrane. Accordingly, sodium extrusion occurs via an ATP-dependent primary sodium pump, in contrast to the Na⁺/H⁺ antiport in most cyanobacteria. A. platensis is strongly dependent on sodium/bicarbonate symport for the uptake of inorganic carbon. Sodium extrusion in the presence of the Photosystem II inhibitor diuron indicates that a significant amount of ATP is supplied by cyclic electron transport around Photosystem I, the content of which in A. platensis is exceptionally high. Plastoquinol is oxidized by two parallel pathways, via the cytochrome b ₆ f complex and a putative cytochrome bd complex, both of which are active in the light and in the dark. This revised version was published online in August 2006 with corrections to the Cover Date.     

鄂尔多斯高原碱湖的钝顶螺旋藻光合生理研究

鄂尔多斯高原碱湖的钝顶螺旋藻光合色素含量高低排列为藻胆素>叶绿素a>类胡萝卜素;各色素具有特定的吸收光谱,活体吸收光谱体现出了各色素的吸收;各色素的荧光发射主峰波长约长于活体的13￣35nm,相对荧光强度约是活体的11倍。其光合速率的日变化呈单峰曲线,13:00时达到最高;光补偿点为28￣30μmol.m-2.s-1;光饱和点为220￣235μmol.m-2.s-1;光合作用的最适温度为35℃。呼吸速率日变化随温度的升高呈缓慢上升的趋势。     

Simultaneous use of urea and potassium nitrate for Arthrospira (Spirulina) platensis cultivation

Urea has been considered as a promising alternative nitrogen source for the cultivation of Arthrospira platensis if it is possible to avoid ammonia toxicity; however, this procedure can lead to periods of nitrogen shortage. This study shows that the addition of potassium nitrate, which acts as a nitrogen reservoir, to cultivations carried out with urea in a fed-batch process can increase the maximum cell concentration (X(m) ) and also cell productivity (P(X) ). Using response surface methodology, the model indicates that the estimated optimum X(m) can be achieved with 17.3 mM potassium nitrate and 8.9 mM urea. Under this condition an X(m) of 6077 ± 199 mg/L and a P(X) of 341.5 ± 19.1 mg L(-1) day(-1) were obtained.     

Purification and characterization of phycoferritin from the blue-green alga, Arthrospira (Spirulina) platensis

Phycoferritin from the nutritionally important blue-green alga Arthrospira platensis has been isolated, by application of conventional biochemical techniques. The molecular mass, yield, iron and total neutral carbohydrate contents of the purified protein were 470 kDa, 0.044 mg g⁻¹ of Arthrospira, 1.4 and 20%, respectively. The iron content was much lower when compared to bacterial and mammalian ferritins. The P: Fe ratio of Arthrospira phycoferritin was 1: 3.5, a value akin to bacterioferritins. Native gel-electrophoresis revealed the presence of isoforms. Subunit analysis by SDS-PAGE and Western blotting showed a protein subunit with an apparent molecular mass of 18 kDa. Oligomeric forms of the protein subunit were also present. The phycoferritin exhibited cross-reactivity with anti-pea seed ferritin suggesting phylogenetic relationship with that of higher plants. Carbohydrate analysis of phycoferritin by GC-MS revealed the presence of sugars such as galactose, glucose and mannose similar to that of mammalian ferritins. Interestingly, the analysis also revealed sugars such as rhamnose, xylose and talose, which has not been reported in the structure of ferritins. Except for very low histidine content in phycoferritin, the rest of the amino acid composition resembled to ferritins of other species. UV-visible spectral analysis of the phycoferritin revealed the presence of haem groups, a property characteristic of bacterioferritins. The fluorescence intensity of phycoferritin was higher than equine spleen ferritin. Circular dichroic spectra revealed a lower degree of helicity.     

Analysis of the promoter region of the RuBisCO gene of Arthrospira (Spirulina) platensis

Ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO), a key enzyme involved with photosynthetic carbon assimilation, catalyzes the carboxylation and oxidization of ribulose-1,5-bisphosphate. Interestingly, the promoter region of this gene from Arthrospira platensis could drive the expression of a downstream gene in Escherichia coli. In this study, using green fluorescent protein as a reporter of gene expression, the structure and function of the promoter region of the RuBisCO gene of A. platensis was analyzed. There are three hypothetical promoter elements predicted in the 200 bp upstream of the open reading frame (ORF) of RuBisCO. Through deletion analysis of the promoter, it was demonstrated that one of these elements was the active promoter, which was located between ?94 and the ORF of RuBisCO. The ?35 box of the RuBisCO promoter (TTGACT) was very similar to that of the rpoD gene (TTGACA) of E. coli, with only the sixth nucleotide divergent. Site-directed mutational analysis showed that when the sixth nucleotide (T) was changed to A, the activity of the promoter remained unchanged. However, when the first and second Ts were mutated, the activities of the promoters decreased drastically. Determining the structure and function of promoters would help elucidate the molecular mechanisms of the gene expression and regulation.     

Effects of light intensity and dilution rate on the semicontinuous cultivation of Arthrospira (Spirulina) platensis. A kinetic Monod-type approach

Semicontinuous cultures were carried out at different dilution rates (D) and light intensities (I) to determine the maximum productivity of Arthrospira platensis cultivated in helicoidal photobioreactor up to the achievement of pseudo-steady-state conditions. At I=108 μmol photons m(-2) s(-1), the semicontinuous regime ensured the highest values of maximum cell concentration (X(m)=5772±113 mg L(-1)) and productivity (P(XS)=1319±25 mg L(-1) d(-1)) at the lowest (D=0.1 day(-1)) and the highest (D=0.3 day(-1)) dilution rates, respectively. A kinetic model derived from that of Monod was proposed to determine the relationship between the product of light intensity to dilution rate (ID) and the cell productivity, which were shown to exert a combined influence on this parameter. This result put into evidence that pseudo-steady-state conditions could be modified according to circumstances, conveniently varying one or other of the two independent variables.     

Polyhydroxyalkanoates and pigments coproduction by Arthrospira (Spirulina) platensis cultivated in crude glycerol

Journal of Applied Phycology - Valorization of industrial waste as an alternative carbon source for microalgae mixotrophic cultivation and co-production of high-value compounds are strategies to...     

Repeated fed-batch cultivation of Arthrospira (Spirulina) platensis using urea as nitrogen source

Arthrospira (Spirulina) platensis (Nordstedt) Gomont was autotrophically cultivated for biomass production in repeated fed-batch process using urea as nitrogen source, with the aim of making large-scale production easier, increasing cell productivity and then reducing the production costs. It was investigated the influence of the ratio of renewed volume to total volume (R), the urea feeding time (t_f) and the number of successive repeated fed-batch cycles on the maximum cell concentration (X_m), cell productivity (P_x), nitrogen-to-cell conversion yield (Y_x/n), maximum specific growth rate (μ_m) and protein content of dry biomass. The experimental results demonstrated that R = 0.80 and t_f = 6 d were the best cultivation conditions, being able to simultaneously ensure, throughout the three fed-batch cycles, the highest average values of three of the five responses (X_m = 2101 ± 113 mg L^?1, P_x = 219 ± 13 mg L^?1 d^?1 and Y_x/n = 10.3 ± 0.8 g g^?1).     

Growth inhibition of the cyanobacterium Spirulina (Arthrospira) platensis by 1.7 MHz ultrasonic irradiation

Ultrasonic waves of high frequency (1.7 MHz) and low intensity (0.6 W cm^–2) were employed to prevent cyanobacterial cells from growing fast and the effects of this growth inhibition were investigated. At least five minutes of ultrasonic irradiation was essential for effective inhibition. The growth rate of irradiated cells was reduced to 38.9% of the control during short-term culture. Longer exposure did not significantly enhance the inhibition. For a particular level of energy input, distributed ultrasonic exposure (more short intermittent exposures) was more effective in inhibiting growth than fewer, but longer exposures. For instance, the final biomass decreased to 30.1% of the control after ultrasonic irradiation for 4 minutes every 3 days, whereas it only decreased to 60% of the control with exposure for 12 minutes every 11 days. It is suggested that distributed ultrasonic irradiation is a practical method to prevent cyanobacterial cells from fast growth. A possible explanation for the inhibition is discussed in relation to cell structure, the absorption spectrum of intact cells, chlorophyll level and oxygen evolution.     

Application of Arthrospira (Spirulina) platensis biomass for silver removal from aqueous solutions

The cyanobacterium Arthrospira (Spirulina) platensis was used to study the process of silver biosorption. Effects of various parameters such as contact time, dosage of biosorbent, initial pH, temperature, and initial concentration of Ag(I) were investigated for a batch adsorption system. The optimal biosorption conditions were determined as pH 5.0, biosorbent dosage of 0.4 g, and initial silver concentration of 30 mg/L. Equilibrium adsorption data were analyzed by the Langmuir and Freundlich models – however, the Freundlich model provided a better fit to the experimental data. The kinetic data fit the pseudo-second-order model well, with a correlation coefficient of 0.99. The analysis of thermodynamic parameters (ΔG°, ΔH° and ΔS°) revealed that the adsorption process of silver ion by spirulina biomass was exothermic and spontaneous (ΔG° < 0), and exothermic (ΔH° < 0) process. The biosorption capacity of biomass A. platensis serves as a basis for the development of green technology for environmental remediation.     

Optimum growth conditions and light utilization efficiency of Spirulina platensis (= Arthrospira fusiformis) (Cyanophyta) from Lake Chitu, Ethiopia

Spirulina platensis (= Arthrospira fusiformis) was isolated from Lake Chitu, a saline, alkaline lake in Ethiopia, where it forms an almost unialgal population. Optimum growth conditions were studied in a turbidostat. Cultures grown in modified Zarrouk's medium and exposed to a range of light intensities (20–500 µmol photons m^–2s^–1) showed a maximum specific growth rate (µ_max) of 1.78 d^–1. Quantum yield for growth (µ) was 3.8% at the optimum light for growth of 330 µmol photons m^–2s^–1, and ranged from 2.8 to 9.4%. With increase in irradiance, the chlorophyll a concentration decreased, and the carotenoids/chlorophyll a ratio increased by a factor of 2.4. The phosphorus to carbon ratio (P/C) showed some variation, while the nitrogen to carbon ratio (N/C) remained relatively constant, thus causing fluctuations in the N:P ratio (7–11) of cells. An optimum N:P ratio of about 7 was attained in cells growing at the optimum light for growth. Results from the continuous culture experiments agreed well with maximum values of photosynthetic efficiency given in the literature for natural populations of S. platensis in the soda lakes of East Africa, Lake Arenguade (Ethiopia), and Lake Simbi (Kenya).     

Induction of axenic culture of Arthrospira (Spirulina) platensis based on antibiotic sensitivity of contaminating bacteria

Arthrospira platensis SAG 21.99 and the isolated bacteria (Halomonas spp., Staphylococcus sp., etc.) from the culture of A. platensis SAG 21.99 were treated with five antibiotics to determine the minimal lethal concentrations. The combination of a washing step and a consecutive treatment with antibiotics, imipenem (100 μg ml⁻¹), neomycin (100 μg ml⁻¹) and cycloheximide (20 μg ml⁻¹), treatment step was highly effective in eliminating bacteria. An axenic culture of A. platensis SAG 21.99 could be induced within 3 days using this method. This technique is a simple and rapid method for obtaining axenic cultures of filamentous cyanobacteria.     

Characterization of the iron-containing superoxide dismutase and its response to stress in cyanobacterium Spirulina (Arthrospira) platensis

Superoxide dismutase (SOD) is considered a primary antioxidant which defends against reactive oxygen species that are induced by environmental stress. In this study, we examined changes in SOD activity and expression in the cyanobacterium Spirulina (Arthrospira) platensis under iron and salinity stress; we characterized its induction under these stress conditions and we overexpressed the enzyme in a bacterial host for preliminary characterization. Analysis of SOD isoforms concludes that S. platensis was found to regulate only the iron-containing SOD isoform (FeSOD) in response to two types of stress that were tested. The FeSOD expression (on the level of both mRNA and enzyme activity) was induced by the stress conditions of salinity and iron levels. The FeSOD from S. platensis was overexpressed in Escherichia coli BL21. The recombinant FeSOD protein (about 23 kDa) was purified for characterization. It showed high specific activity and pH stability at 6.0–9.0, and it is relatively thermostable, retaining 45 % of its activity after 30 min at 90 °C. Phylogenetic analysis reveals that S. platensis FeSOD is grouped with the FeSODs from other cyanobacterial species and separated from those of the eukaryotic Chlorophyta, suggesting that the FeSOD gene may be used as a molecular marker in physiological, phylogenetic, and taxonomic studies. This study also suggests that the increased activity and expression of SOD may play a role in algal survival under stress conditions.