Biomass and nutritive value of Spirulina (Arthrospira fusiformis) cultivated in a cost-effective medium

Introduction

Cultivation of spirulina at commercial-scales relies on analytical grade–based media, which are expensive and so are the product.

Purpose

This study assessed the biomass, proximate composition, and other useful compounds in Spirulina (Arthrospira fusiformis) produced with a cost-effective culture medium (LCMA), and the results were compared with those from a standard Zarrouk medium–grown spirulina.

Methods

The LCMA medium was formulated by using a commercial NPK10-20-20 fertilizer as a source of the three major nutrients for spirulina growth, and other three ingredients from Zarrouk medium. The experiment was conducted for 28 days in the glass aquaria under indoor conditions. Standard analytical methods were applied for the determination of proximate composition, chlorophyll, minerals, and vitamins in the spirulina biomass.

Result

The LCMA medium showed the best growth conditions by accumulating higher chlorophyll content (0.99 ± 0.02%) and dry weight (0.75 ± 0.01 g/100 ml) as well as attaining higher optical density (2.06 at day 15) earlier than the Zarrouk medium. The results of the proximate analysis for spirulina cultured in the LCMA medium were of good quality, with the protein contributing more than 50% of its dry matter. It was further noticed that the LCMA was an ideal medium for optimization of vitamins and some minerals since it recorded a significant amount of most of the analyzed vitamins together with the minerals sodium and potassium compared with the Zarrouk medium.

Conclusion

It is suggested that LCMA medium could be used as the alternative and cheap medium for maximization of biomass and production of useful biochemical compounds in spirulina species.

     

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.     

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).     

Response of Spirulina platensis (= Arthrospira fusiformis) from Lake Chitu, Ethiopia, to salinity stress from sodium salts

Spirulina platensis (=Arthrospira fusiformis) was isolated from Lake Chitu, a soda crater lake in the Ethiopian Rift Valley, where it formsa dense and almost unialgal population. Growth experiments were run in turbidostats under constant light, to assess growth response and tolerance to salinity, as well as to the component anions. Salinity was tested over the range 13–88 g L^-1 using additions of NaHCO3, NaCl or Na2SO4. A maximum specific growth rate (μ_max d^-1) of 2.14 was achieved at the lowest salinity, but quantum yield (Φ%) was highest between 33 to 51 g L^-1. Increasing salinity of the medium reduced the specific growth rate (μ) to a minimum of 0.33 d^-1, and Φ to < 0.5%. Growth response in terms of μ and Φ was best in HCO ₃ ^- , less in Cl^-, and least in SO ₄ ^2- series. Cultures showed obvious differences in cellular morphology, pigment, nitrogen and phosphorus contents in response to treatment with the different anions. Results indicate that the species has a wide range of tolerance to salinity from NaHCO3. Some degree of tolerance is also shown to high concentrations of Cl^- and SO ₄ ² , but with an overall lower performance of cells in terms of growth rate, light utilization efficiency, and nutrient status to cells grown in high HCO ₃ ^- concentrations and the same levels of salinity and light. This revised version was published online in August 2006 with corrections to the Cover Date.     

Morphological variability of Arthrospira (Spirulina) fusiformis (Cyanophyta) in relation to environmental variables in the tropical soda lake Chitu,Ethiopia

Although variability of Arthrospira morphology, widely used in taxonomy and other valuable aspects, as a response to changes in environmental variables has been studied in laboratory cultures, little is known about such responses in the natural habitats. In this study, Arthrospira fusiformis was characterized morphologically in relation to selected environmental variables in the natural ecosystem. Three distinct morphotypes, tightly coiled (H-type), spiral or loosely coiled (S-type), and intermediately coiled (C-type) were observed and described. They varied largely in the degree of coiling, trichome and helix dimensions, coils, shape of trichome end and trichome abundance. The H-type was the most dominant (50%) followed by the S-type (40%). The dominance of H-type and tightening of helix pitch were strongly associated with NO₃ ^? and HCO₃ ^? deficiencies, and high levels of photosynthetically active radiation (PAR) and temperature of the lake. The abundance of S-type was more strongly but negatively related to the high salinity of the lake, probably indicating that decreasing salinity may favor the S-type. In conclusion, the variability in abundance and morphology seem to suggest that A. fusiformis could show morphological modifications in response to environmental stresses in their natural soda lakes, resulting in occurrence of various morphotypes of the same species.     

Evaluation of growth and biomass production of Arthrospira (Spirulina) fusiformis in laboratory cultures using waters from the Ethiopian soda lakes Chitu and Shala

The cost of nutrient media is the major challenge for biomass production of Spirulina. Although much effort has been made to use enriched seawater for the cultivation of this microalga, little attention has been given to the potential of water of soda lakes. In this study, growth (μ, day^?1) and biomass production (B) of Arthrospira fusiformis cultivated using waters of the soda lakes Chitu and Shala with or without supplementation were evaluated. Comparable μ and B values were achieved in both Lake Chitu water-based media (CBM) and Lake Shala water-based media (SBM), with slightly higher values in the latter. Both CBM and SBM supplemented with the standard Spirulina medium (SM) by 25 % and 50 % supported considerably higher μ and B. The pH and salinity of the cultures showed significant variations (P?μ and B. The observed higher μ and B were probably associated with the reduction in pH and salinity of the supplemented media due to addition of bicarbonate–carbonates and dilution, and provision of the limiting nutrient nitrogen. The higher μ and B in SBM may have resulted from some of their aggregate chemical parameters, which were closer to those in the SM, and abundant PO₄-P. This seems to suggest that Lake Shala water is more conducive to Arthrospira. We contend that 25 % and 50 % supplemented Lake Shala water can be preferably used to produce Arthrospira biomass, thereby reducing the cost of nutrients by 75 % and 50 %, respectively.     

Purification and immunomodulating activity of C-phycocyanin from Spirulina platensis cultured using power plant flue gas

In this study, flue gas from a power plant smokestack was applied to culture Spirulina platensis microalgae. Our results will not only achieve the fixation of carbon from the emissions, products can also be produced from the algal biomass that possess physiological activities which could be beneficial to human health. An improved one-step process of chromatography was used to produce high-purity C-phycocyanin with a PC ratios >3.5. Adding different concentrations of ammonium sulfate produced different amounts of C-phycocyanin, with 40% generating the highest yield, followed by 35% and 30% concentrations. Immunomodulating activities were evaluated in the murine macrophage cell line J774A.1. We found that C-phycocyanin had the capability to induce secretion of inflammatory cytokines, including TNF-α, IL-1β, and IL-6, and that these results were not due to contamination with LPS. Treatment with C-phycocyanin also increased proIL-1β and COX-2 protein expression dose-dependently. Furthermore, C-phycocyanin rapidly stimulated phosphorylation of inflammatory-related signaling molecules, including ERK, JNK, p38 and IκB. In addition, although C-phycocyanin decreased production of LPS-induced ROS, it did not inhibit LPS-induced inflammatory cytokines in J774A.1 cells. This is the first report to show that C-phycocyanin exhibited a detailed molecular mechanism of bioactivity by boosting immunomodulation performance.     

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.     

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.     

Spirulina (Arthrospira): potential application as an animal feed supplement

Spirulina has been used as human food supplement for over 20 years, but its use as an animal feed supplement is relatively recent. This paper reviews earlier studies on its more established effects on growth, survival and tissue quality in a whole range of animals and more recent studies on its immunomodulatory, anti-viral and anti-cancer effects. The basis for potential application as an animal feed supplement is discussed.     

Crystal structure of a light-harvesting protein C-phycocyanin from Spirulina platensis

The crystal structure of C-phycocyanin, a light-harvesting phycobiliprotein from cyanobacteria (blue-green algae) Spirulina platensis has been solved by molecular replacement technique. The crystals belong to space group P2(1) with cell parameters a = 107.20, b = 115.40, c = 183.04 A; beta = 90.2 degrees. The structure has been refined to a crystallographic R factor of 19.2% (R(free) = 23.9%) using the X-ray diffraction data extending up to 2.2 A resolution. The asymmetric unit of the crystal cell consists of two (alphabeta)6-hexamers, each hexamer being the functional unit in the native antenna rod of cyanobacteria. The molecular structure resembles that of other reported C-phycocyanins. However, the unique form of aggregation of two (alphabeta)6-hexamers in the crystal asymmetric unit, suggests additional pathways of energy transfer in lateral direction between the adjacent hexamers involving beta155 phycocyanobilin chromophores.     

Interaction of Pb(II) ions with C-phycocyanin from Spirulina platensis: effect of ionic strength

The effect of Pb(II) ions on C-phycocyanin from Spirulina platensis was studied by the method of fluorescence spectroscopy. The efficiency of quenching was analyzed using the Stern-Volmer relation. Isotherms of adsorption were constructed, and the constants of binding of Pb(II) ions to C-phycocyanin in solutions of varying ionic strength were determined by equilibrium dialysis and atomic absorption spectroscopy. The constants of binding of Pb(II) ions to C-phycocyanin in 2, 20, and 50 mM NaN03 solutions were estimated to be 4.79 x 10(5), 3.63 x 10(5) and 2.82 x 10(5), respectively. It was found that the interaction of Pb(II) ions with C-phycocyanin has a cooperative character at all values of ionic strengths studied.     

Binding of Cd(II) ions by C-phycocyanin during Spirulina platensis cell growth

Modern effective methods of preparative biochemistry were used to obtain a highly purified preparation of the globular protein C-phycocyanin from Cyanophyceae strain IPPAS B-256. It was shown that C-phycocyanin accumulates endogeneous Cd(II) during the growth of Spirulina platensis cells.     

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).     

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.     

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。     

Physiological behaviour of Arthrospira (Spirulina) maxima during acclimation to changes in irradiance

The physiological behaviour of Arthrospira (Spirulina) maxima during acclimation to sudden changes in irradiance from high (HL) to low light (LL) and vice versa was studied by following parameters concerning growth rate, pigment, carbohydrate and protein cell contents. Applying first order kinetics, the specific acclimation rates for the parameters considered were calculated. During HL to LL shift, pigments increased to compensate for a reduction in growth irradiance in order to maintain relatively high growth rates, whereas carbohydrates decreased at the highest rate. The synthesis of phycobiliproteins proceeded at a rate similar or little higher than that of chlorophyll a, indicating their importance in the light harvesting at low irradiance. During LL to HL shift, carbohydrate biosynthesis was increased, whereas pigment and protein cell contents decreased. The kinetic analysis suggested that the pigment decrease could be accounted for both by dilution through growth and in vivo degradation. During this transition, the initially high cell pigment content gave rise to a very heavy carbohydrate synthesis, which for a short time, after the shift to HL conditions, overshot the final steady-state. In the same period the specific growth rate also increased notably, overshooting the μmax. The acclimation rates of the measured parameters were faster during LL to HL transition then during the reverse. The physiological response of A. maxima during the acclimation to sudden irradiance shifts points out the ability of this cyanobacterium to alter light harvesting and highlights again the key role of carbohydrates when the cells underwent an energy crisis during down-shift. This revised version was published online in August 2006 with corrections to the Cover Date.     

Large-scale recovery of C-phycocyanin from Spirulina platensis using expanded bed adsorption chromatography

C-phycocyanin was purified on a large scale by a combination of expanded bed adsorption, anion-exchange chromatography and hydroxyapatite chromatography from inferior Spirulina platensis that cannot be used for human consumption. First, phycobiliproteins were extracted by a simple, scaleable method and then were recovered by Phenyl-Sepharose chromatography in an expanded bed column. The purity (the A(620)/A(280) ratio) of C-phycocyanin isolated with STREAMLINE column was up to 2.87, and the yield was as high as 31 mg/g of dried S. platensis. After the first step, we used conventional anion-exchange chromatography for the purification steps, with a yield of 7.7 mg/g of dried S. platensis at a purity greater than 3.2 and with an A(620)/A(650) index higher than 5.0. The fractions from anion-exchange chromatography with a level of purity that did not conform to the above standard were subjected to hydroxyapatite chromatography, with a C-PC yield of 4.45 mg/g of dried S. platensis with a purity greater than 3.2. The protein from both purification methods showed one absolute absorption peak at 620 nm and a fluorescence maximum at 650 nm, which is consistent with the typical spectrum of C-phycocyanin. SDS-PAGE gave two bands corresponding to 21 and 18 kDa. In-gel digestion and LC-ESI-MS showed that the protein is C-phycocyanin.     

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.     

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...