Nutritional optimization of Arthrospira platensis for starch and Total carbohydrates production.

Present study aims to optimize the production of starch and total carbohydrates from Arthrospira platensis. Growing concerns toward unprecedented environmental issues associated with plastic pollution has created a tremendous impetus to develop new biomaterials for the production of bioplastic. Starch-based biopolymers from algae serve as sustainable feedstock for thermoplastic starch production due to their abundant availability and low cost. A. platensis was cultivated in Zarrouk's medium at 32 ± 1°C and exposed to red light with a photoperiod of 12:12 hr light/dark. Growth kinetics studies showed that the maximum specific growth rate (µmax ) obtained was 0.059 day-1 with the doubling time (td ) of 11.748 days. Subsequently, Zarrouk's medium with different concentrations of sulfur, phosphorus and nitrogen was prepared to establish the nutrient-limiting conditions to enhance the accumulation of starch and total carbohydrates. In this study, the highest starch accumulated was 6.406 ± 0.622 mg L-1 under optimized phosphorus limitation (0.025 g L-1 ) conditions. Nitrogen limitation (0.250 g L-1 ) results demonstrated significant influenced (p < 0.05) on total carbohydrates (67.573 ± 2.893 mg L-1 ) accumulation in A. platensis. The starch accumulation in A. platensis was significantly affected (p < 0.05) by phosphorus limitation (0.0025 g L-1 ). Subsequently, the optimized phosphorus concentration was coupled with mixotrophic cultivation to further enhance the starch accumulation. The results obtained indicated that, the starch (11.426 ± 0.314 mg L-1 ) and carbohydrates (43.053 ± 2.986 mg L-1 ) concentration obtained was significantly high (p < 0.05) under mixotrophic cultivation. Therefore, it shown that nutrient limitation and mixotrophic cultivation are viable strategies to enhance the accumulation of starch and total carbohydrates in A. platensis.

Performance of the mixed LED light quality on the growth and energy efficiency of Arthrospira platensis.

The effect of mixed light quality with red, blue, and green LED lamps on the growth of Arthrospira platensis was studied, so as to lay the theoretical and technical basis for establishing a photo-bioreactor lighting system for application in space. Meanwhile, indexes, like morphology, growth rate, photosynthetic pigment compositions, energy efficiency, and main nutritional components, were measured respectively. The results showed that the blue light combined with red light could decrease the tightness of filament, and the effect of green light was opposite. The combination of blue light or green light with red light induced the filaments to get shorter in length. The 8R2B treatment could promote the growth of Arthrospira platensis significantly, and its dry weight reached 1.36 g L-1, which was 25.93% higher than the control. What's more, 8R2B treatment had the highest contents of carbohydrate and lipid, while 8R2G was rich in protein. 8R0.5G1.5B had the highest efficiency of biomass production, which was 161.53 mg L-1 kW-1 h-1. Therefore, the combination of red and blue light is more conducive to the growth of Arthrospira platensis, and a higher biomass production and energy utilization efficiency can be achieved simultaneously under the mixed light quality with the ratio of 8R0.5G1.5B.

Effect of Ultrasonic Extraction on Production and Structural Changes of C-Phycocyanin from Marine Spirulina maxima.

This work first showed that very high amounts of phycocyanins, such as 11.3 mg/mL C-phycocyanin (C-PC), 3.1 mg/mL allophycocyanin (APC), and 0.8 mg/mL phycoerythrin (PE), can be obtained using an ultrasonic extraction process (UE) with a 60 kHz frequency and 3 h of process time at 25 °C, without any other pretreatments. These yields were higher than those from most conventional water extractions at 4 °C for 24 h (Control condition) or at 25 °C for 24 h (WE), namely, 9.8 and 5.7 mg/mL C-PC, 2.3 and 1.2 mg/mL APC, and 0.7 and 0.3 mg/mL PE, respectively. These yields were also shown to be even higher than yields from other reported data. Structural changes in C-PC in the extracts were also found for the first time, according to extraction conditions, showing that the total concentration of C-PC and of the α-subunit of C-PC in the UE were much higher than in the WE, with little difference in the amount of ß-subunit of C-PC in the UE or WE. It was also shown that the structural changes in C-PC in the WE decreased both antioxidant and anti-inflammation activities-29.83% vs. 32.09% of α,α-diphenyl-ß-picrylhydrazyl (DPPH) scavenging activity and 8.21 vs. 7.25 µM of NO production for the WE and UE, respectively-while the UE, with similar patterns to standard C-PC, showed very high biological effects, which may suggest that the biologically active part is the α-subunit of C-PC, not the ß-subunit.

Composition and functional property of photosynthetic pigments under circadian rhythm in the cyanobacterium Spirulina platensis.

Circadian rhythm is an important endogenous biological signal for sustainable growth and development of cyanobacteria in natural ecosystems. Circadian effects of photosynthetically active radiation (PAR), ultraviolet-A (UV-A) and ultraviolet-B (UV-B) radiations on pigment composition have been studied in the cyanobacterium Spirulina platensis under light (L)/dark (D) oscillation with a combination of 4/20, 8/16, 12/12, 16/8, 20/4 and 24/24 h time duration. Circadian exposure of PAR + UV-A (PA) and PAR + UV-A + UV-B (PAB) showed more than twofold decline in Chl a, total protein and phycocyanin (PC) in light phase and significant recovery was achieved in dark phase. The fluorescence emission wavelength of PC was shifted towards lower wavelengths in the light phase of PAB in comparison to P and PA whereas the same wavelength was retrieved in the dark phase. The production of free radicals was accelerated twofold in the light phase (24 h L) whereas the same was retrieved to the level of control during the dark phase. Oxidatively induced damage was alleviated by antioxidative enzymes such as catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and ascorbate peroxidase (APX) in the light phase (0-24-h L) whereas the dark phase showed significant inhibition of the same enzymes. Similar characteristic inhibition of free radicals and recovery of PC was observed inside cellular filament after circadian rhythm of 24/24 h (L/D). Circadian exposure of P, PA and PAB significantly altered the synthesis and recovery of pigments that could be crucial for optimization and sustainable production of photosynthetic products for human welfare.

Dairy farm wastewater treatment and lipid accumulation by Arthrospira platensis.

Dairy cattle treated wastewaters are potential resources for production of microalgae biofuels. A study was conducted to evaluate the capability of Arthrospira platensis cultivated in dairy farm wastewater for biodiesel production. The biomass of Arthrospira platensis was found to be 4.98 g L-1 and produced 30.23 wt% lipids to dry biomass cultivated in wastewater which was found nitrogen stressed in photo bioreactor. The extracted lipid displayed a suitable fatty acid profile for biodiesel, although the content of linolenic acid was found a little higher than the standard EN14214. It was found that nitrogen stressed medium increase the total lipid content but temperature and intensities of light were the most important factors to control the quantity of linolenic acid and hence the quality of biodiesel, while the optimum CO2 helped to achieve maximum biomass and triacylglycerols. The Arthrospira platensis offer a good option for the treatment of wastewater before final discharge.

Higher adsorption capacity of Spirulina platensis alga for Cr(VI) ions removal: parameter optimisation, equilibrium, kinetic and thermodynamic predictions.

This study discusses about the biosorption of Cr(VI) ion from aqueous solution using ultrasonic assisted Spirulina platensis (UASP). The prepared UASP biosorbent was characterised by Fourier transform infrared spectroscopy, X-ray diffraction, Brunauer-Emmet-Teller, scanning electron spectroscopy and energy dispersive X-ray and thermogravimetric analyses. The optimum condition for the maximum removal of Cr(VI) ions for an initial concentration of 50 mg/l by UASP was measured as: adsorbent dose of 1 g/l, pH of 3.0, contact time of 30 min and temperature of 303 K. Adsorption isotherm, kinetics and thermodynamic parameters were calculated. Freundlich model provided the best results for the removal of Cr(VI) ions by UASP. The adsorption kinetics of Cr(VI) ions onto UASP showed that the pseudo-first-order model was well in line with the experimental data. In the thermodynamic study, the parameters like Gibb's free energy, enthalpy and entropy changes were evaluated. This result explains that the adsorption of Cr(VI) ions onto the UASP was exothermic and spontaneous in nature. Desorption of the biosorbent was done using different desorbing agents in which NaOH gave the best result. The prepared material showed higher affinity for the removal of Cr(VI) ions and this may be an alternative material to the existing commercial adsorbents.

Biochemical composition and antioxidant activities of Arthrospira (Spirulina) platensis in response to gamma irradiation.

Arthrospira (Spirulina) platensis is a blue-green alga, rich with bioactive components and nutrients. To evaluate effect of gamma irradiation, A. platensis was exposed to different doses of 0.0, 0.5, 1.0, 1.5, 2.0 and 2.5kGy. The data showed that the phenolic and proline contents significantly increased with the increase of gamma irradiation doses up to 2.0kGy, above which a reduction was observed. The soluble proteins and malondialdehyde (MDA) contents were stimulated by all tested irradiation doses. Furthermore, the vitamins (A, K and B group) and mineral contents (N, P, Na, K, Ca, Mg and Fe) were stimulated by the irradiation doses compared with the control. The activities of some N-assimilating and antioxidant enzymes were significantly increased with the irradiation doses up to 2.0kGy. This study suggests the possible use of gamma irradiation as a stimulatory agent to raise the nutritive value and antioxidant activity of A. platensis.

Mass cultivation of UV-B adapted Arthrospira platensis RRGK under open raceway pond for the production of Poly-ß-hydroxy butyrate.

Six different strains of cyanobacteria were isolated from the freshwater lake, Arakkonam, India. Staining of cells with Nile Red showed the presence of large quantities of PHB granules in the cell cytoplasm of Arthrospira. Molecular identification of the strain was carried out using 16S rRNA analysis and their systematic position was ascertained as Arthrospira platensis RRGK. Studies were carried out on random mutagenesis approach using UV-B radiation for enhancing the production of PHB. Further, Response Surface Methodology was used for optimization of pH, temperature, and sodium bicarbonate for higher biomass and PHB production. Under open raceway pond A. platensis RRGK produced biomass concentration of 2.2±0.13gL-1 and 131±0.36gL-1 of PHB content. It was chemically characterized through FTIR, DSC, TGA and XRD analyses. Hence, PHB can be produced from cyanobacteria by sequestering harmful CO2. It can also be used as a substitute for synthetic polymers in tissue engineering.

Adenylate cyclase in Arthrospira platensis responds to light through transcription.

Cyclic 3',5' adenosine monophosphate (cAMP) is a ubiquitous signaling molecule, but its role in higher plants was in doubt due to its very low concentration. In this study we wanted to look at the flux of cAMP in response to light in algae, considered to be the more primitive form of photosynthetic organisms. While it did not fluctuate very much in the tested green algae, in the cyanobacterium Arthrospira platensis its level was closely linked to exposure to light. The expression from cyaC, the major isoform of adenylate cyclase was strongly influenced by exposure of the cells to light. There was about 300 fold enhancement of cyaC transcripts in cells exposed to light compared to the transcripts in cells in the dark. Although post-translational regulation of adenylate cyclase activity has been widely known, our studies suggest that transcriptional control could also be an important aspect of its regulation in A. platensis.

Effect of light intensity on the degree of ammonia toxicity on PSII activity of Arthrospira platensis and Chlorella vulgaris.

Herein the effect of increasing light intensity on the degree of ammonia toxicity and its impact on the photosynthetic performance of Arthrospira and Chlorella was investigated using Chl fluorescence as a technique to characterize their photosystem II (PSII) activity. The results revealed that the increase of light intensity amplifies the ammonia toxicity on PSII. Chl fluorescence transients shown that at a given free ammonia (FA) concentration (100mg-N/L), the photochemistry potential decreased by increasing light intensity. The inhibition of the PSII was not reversible either by re-incubating the cells under dark or under decreased FA concentration. Moreover, the decrease of photochemical and non-photochemical quenching (NPQ) of fluorescence suggest that ammonia toxicity decreases the open available PSII centers, as well the inability of PSII to transfer the generated electrons beyond QA. The collapse of NPQ suggests that ammonia toxicity inhibits the photoprotection mechanism(s) and hence renders PSII more sensitive to photoinhibition.

A new bioenergetic and thermodynamic approach to batch photoautotrophic growth of Arthrospira (Spirulina) platensis in different photobioreactors and under different light conditions.

Photobioreactor configuration, mode of operation and light intensity are known to strongly impact on cyanobacteria growth. To shed light on these issues, kinetic, bioenergetic and thermodynamic parameters of batch Arthrospira platensis cultures were estimated along the time at photosynthetic photon flux density (PPFD) of 70µmolm(-2)s(-1) in different photobioreactors with different surface/volume ratio (S/V), namely open pond (0.25cm(-1)), shaken flask (0.48cm(-1)), horizontal photobioreactor (HoP) (1.94cm(-1)) and helicoidal photobioreactor (HeP) (3.88cm(-1)). Maximum biomass concentration and productivity remarkably increased with S/V up to 1.94cm(-1). HoP was shown to be the best-performing system throughout the whole runs, while HeP behaved better only at the start. Runs carried out in HoP increasing PPFD from 40 to 100µmolm(-2)s(-1) revealed a progressive enhancement of bioenergetics and thermodynamics likely because of favorable light distribution. HoP appeared to be a promising configuration to perform high-yield indoor cyanobacterial cultures.

Impact of natural light on growth and biopigment profile of cyanobacteria Spirulina platensis.

Cyanobacteria are economically important microorganisms and good source of natural pigments such as chlorophyll, carotenoids and phycobilliproteins. The present research work showed the optimum combination of photophase and scotophase of Spirulina platensis on biomass and chlorophyll-a, carotenoids, phycocyanin, allophycocyanin, and phycoerythrin contents. The study revealed that among all six light conditions tested, the cultures placed at west facing window, receiving natural day light at temperature 30°C showed extremely significant higher biomass (O.D. 3.46 ± 0.17%) and biopigment accumulation Chlorophyll a 8.94 ± 0.43%, Carotenoid 1.62 ± 0.18%, phycocyanin 2.26 ± 0.14%, allophycocyanin 2.66 ± 0.18% and phycoerythrin 1.32 ± 0.31% as compared to the standard (Full day natural light), which might be beneficial for large scale production of biopigment.

Temporal Gene Expression of the Cyanobacterium Arthrospira in Response to Gamma Rays.

The edible cyanobacterium Arthrospira is resistant to ionising radiation. The cellular mechanisms underlying this radiation resistance are, however, still largely unknown. Therefore, additional molecular analysis was performed to investigate how these cells can escape from, protect against, or repair the radiation damage. Arthrospira cells were shortly exposed to different doses of 60Co gamma rays and the dynamic response was investigated by monitoring its gene expression and cell physiology at different time points after irradiation. The results revealed a fast switch from an active growth state to a kind of 'survival modus' during which the cells put photosynthesis, carbon and nitrogen assimilation on hold and activate pathways for cellular protection, detoxification, and repair. The higher the radiation dose, the more pronounced this global emergency response is expressed. Genes repressed during early response, suggested a reduction of photosystem II and I activity and reduced tricarboxylic acid (TCA) and Calvin-Benson-Bassham (CBB) cycles, combined with an activation of the pentose phosphate pathway (PPP). For reactive oxygen species detoxification and restoration of the redox balance in Arthrospira cells, the results suggested a powerful contribution of the antioxidant molecule glutathione. The repair mechanisms of Arthrospira cells that were immediately switched on, involve mainly proteases for damaged protein removal, single strand DNA repair and restriction modification systems, while recA was not induced. Additionally, the exposed cells showed significant increased expression of arh genes, coding for a novel group of protein of unknown function, also seen in our previous irradiation studies. This observation confirms our hypothesis that arh genes are key elements in radiation resistance of Arthrospira, requiring further investigation. This study provides new insights into phasic response and the cellular pathways involved in the radiation resistance of microbial cells, in particularly for photosynthetic organisms as the cyanobacterium Arthrospira.

In vivo estimation of pigment composition and optical absorption cross-section by spectroradiometry in four aquatic photosynthetic micro-organisms.

The objective of the present study was to estimate in vivo pigment composition and to retrieve absorption cross-section values, a(∗), of photosynthetic micro-organisms using a non-invasive technique of reflectance spectrometry. To test the methodology, organisms from different taxonomical groups and different pigment composition were used (Spirulina platensis a Cyanophyta, Porphyridium cruentum a Rhodophyta, Dunaliella tertiolecta a Chlorophyta and Entomoneis paludosa a Bacillariophyta) and photoacclimated to two different irradiance levels: 25 µmol photonm(-2)s(-1) (Low Light, LL) and 500 µmol photonm(-2)s(-1) (High Light, HL). Second derivative spectra from reflectance were used to identify pigment in vivo absorption bands that were linked to specific pigments detected by high performance liquid chromatography. Whereas some absorption bands such as those induced by Chlorophyll (Chl) a (416, 440, 625 and around 675 nm) were ubiquous, others were taxonomically specific (e.g. 636 nm for Chl c in E. paludosa) and/or photo-physiological dependent (e.g. 489 nm for zeaxanthin in the HL-acclimated S. platensis). The optical absorption cross-section, a(∗), was retrieved from reflectance data using a radiative transfer model previously developed for microphytobenthos. Despite the cellular Chl a decrease observed from LL to HL (up to 88% for S. platensis), the a(∗) increased, except for P. cruentum. This was attributed to a 'package effect' and to a greater absorption by photoprotective carotenoids that did not contribute to the energy transfer to the core Chl a.

A novel photobioreactor structure using optical fibers as inner light source to fulfill flashing light effects of microalgae.

In this work, a novel photobioreactor structure using optical fibers being fixed vertically to culture flow direction as inner light source was proposed to fulfill flashing light effects (FLE) of microalgae, so as to obtain high light efficiency. Three types of optical-fiber photobioreactor fulfilling FLE of microalgae, i.e. air-driven panel, pump-driven panel and stirred tank type, were proposed and a 130 L airlift panel one was practically constructed on which both cold (light profile, liquid velocity) and hot model tests were carried out. Results demonstrated that it could produce uniformed light/dark frequencies being over 10 Hz and microalgae productivity increased by 43% and 38% for Spirulina platensis and Scenedesmus dimorphus respectively, compared with the control. This suggested the structure to be a viable and promising option for future photobioreactors.

Enhanced photosynthetic output via dichroic beam-sharing.

Microbial solar biofuels offer great promise for future sustainable food, fuels and chemicals but are limited by low productivities and a requirement for large land areas to harvest sunlight. A 71 % increase in combined photosynthetic activity was achieved by illuminating both Rhodobacter sphaeroides and Arthrospira (Spirulina) platensis from a single beam of simulated sunlight, divided using a dichroic mirror. Therefore, this technique is termed 'dichroic beam-sharing', in which the complementary action spectra of two different useful micro-organisms, belonging to green and purple groups, is exploited and allows a single beam of sunlight to be shared efficiently between separate photobioreactors. Because the action spectra of these two organisms are typical of large groups, this novel method could increase the productivity of photosynthetic micro-organisms in the production of diverse commodities.

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.

Kinetics and dynamics for light state transition in cyanobacterium Spirulina platensis cells.

Light state transition in oxygenic organisms was defined as the ability to equalize the excitation of the two photosystems for maximal photosynthetic efficiency. In cyanobacteria, extensive researches on state transition have continuously provided new knowledge in the past decades but the molecular mechanism and physiological significance are still ambiguous. In this work, kinetics and dynamics of the transition from state 1 to state 2 in cyanobacterium Spirulina platensis cells were studied at different intensity of orange light from 10 to 120 µmol m(-2) s(-1). It was revealed that the state transition worked constantly independent of light intensity while the rates varied. The synchronous fluorescence kinetics for phycobilisome (PBS) and photosystem components indicated that the state transition was entirely regulated by "mobile PBS", and continuously changed fluorescence amplitudes suggested a series of intermediate states were involved between state 1 and state 2. The dynamic property of PBS movement during the state transition was revealed by (1,0) distribution of photo-linkable PBSs, indicating a collective movement of all PBSs. The results suggest that state transition in cyanobacteria possesses not only physiological but also photochemical significance.

Design and optimization of photo bioreactor for O2 regulation and control by system dynamics and computer simulation.

In this paper, a valid kinetic model of photo bioreactor (PBR) used for highly-effective cultivation of blue algae, Spirulina platensis, was developed for fully describing the dynamic characteristics of O(2) concentration, then a closed-loop PBR with Linear-Quadratic Gaussian (LQG) servo controller was established and optimized via digital simulation and dynamic response optimization, and the effectiveness of the closed-loop PBR was further tested and accredited by real-time simulation. The result showed that the closed-loop PBR could regulate and control the O(2) concentration in its gas phase according to the reference with desired dynamic response performance, hence microalgae with unique characteristic could be selected as a powerful tool for O(2) regulation and control whenever O(2) concentration in Bioregenerative Life Support System (BLSS) deviates from the nominal level in emergencies, and greatly enhance safety and reliability of BLSS on space and ground missions.

D1 protein turnover is involved in protection of Photosystem II against UV-B induced damage in the cyanobacterium Arthrospira (Spirulina) platensis.

By using two strains of Arthrospira (Spirulina)platensis, an economically important filamentous cyanobacterium, we compared the impairment of PSII activity and loss of D1 protein content under UV-B radiation. Our study showed that UV-B radiation induced a gradual loss of the oxygen-evolving activity to about 56% after 180 min UV-B irradiation both in strains 439 and D-0083, which have been kept under indoor and an outdoor culturing conditions, respectively for a prolonged period of time. The loss of oxygen evolution was accelerated in both strains in the presence of lincomycin, an inhibitor of protein synthesis, and the amount of D1 protein showed a decrease comparable to that of oxygen evolution during the UV-B exposure. However, the UV-B induced loss of oxygen-evolving activity and D1 protein amount was largely prevented when A. platensis cells were exposed to UV-B irradiance supplemented with visible light. Comparison of the two strains also showed a smaller extent of D1 protein synthesis dependent PSII repair in the indoor strain. Our results show that turnover of the D1 protein is an important defense mechanism to counteract the UV-B induced damage of PSII in A. platensis, and also that visible light plays an important role in maintaining the function of PSII under simultaneous exposure to UV-B and visible light.