Content of Phycobilin Pigments in Two Strains of Cyanobacteria of The

www.symbiosisonline.org Symbiosis www.symbiosisonlinepublishing.com Short Communication SOJ Microbiology & Infectious Diseases Open Access Content of Phycobilin Pigments in Two Strains of Cyanobacteria of the Atacama Desert, Chile Iris Pereira1*, Iván Razmilic2 and Jeffrey Johansen3 1Institute of Biological Sciences, Universidad de Talca, Talca, Chile 2Department of Chemistry, Institute of Natural Resources, University of Talca, Talca, Chile, Chile 3Department Biology, John Caroll University, Cleveland, USA Received: February 24, 2014; Accepted: June 24, 2014; Published: June 27, 2014 *Corresponding author: Iris Pereira, Institute of Biological Sciences, Universidad de Talca, 2 Norte 685, Talca, Chile, E-mail: [email protected] The phycobilin pigments have been used successfully in the Abstract location of tumor cells in the treatment of cancer and it is known that the C-phycocyanin ameliorates experimental autoimmune Cyanobacteria with the encephalomyelitis and induces regulatory T-cells [1]. The light The aim of this study was to assess qualitative and quantitatively Thephycobilin studied pigments strains wereof two Nostoc nitrogen-fixing commune and Tolypothrix tenuis purpose to optimize in the future, the culture conditions to high scale. that phycobilins may be used as chemical “tags”. The pigments produced by this fluorescence is so distinctive and reliable, Chile. They were obtained from a total of twelve transects made, which were gotten from cryptogamic crusts of the Atacama Desert, solution of cells. Also these molecules are used in biotechnology [2]are chemicallyand in food bonded due to to its antibodies, antioxidant which activity. are then Based put on into the a min.between After La they Serena were and inoculated Iquique, inin thePetri north plates of Chile.and then Soil samplesisolated of each station were activated in 250 ml Erlenmeyer flasks for 30 they were cultured for 3 weeks or one month in Petri Plates on agar and massed to a low scale. Once the soil samples were activated, obtained information, we proposed to assess qualitative and quantitatively the content of these pigments in two nitrogen- conditionsZ8-N medium and and continuous later the irradiance.inoculates wereChemical transferred determination to 250 mlof fixers cyanobacteria strains from Atacama Desert. Therefore, pigmentsErlenmeyer and flasks their inconcentrations Z8-N liquid medium were made under by spectrophotometry. room temperature withthe following the purpose objectives to optimize were the proposed: conditions a) toof assesstheir culture quali- andin a quantitatively the phycobilin pigments that these strains possess the C-phycocianin. Strain of Tolypothrix high scale. In both strains, C-phycoerythrin was found in bigger proportion than Materials and Methods other hand The Nostoc presents more quantity of phycobilin pigments: 0.66 mg/ml PE and 0.32 mg/ml PC. On the Collection of the Samples strain, showed 0.080 mg/ml PE and 0.037 determinationmg/ml PC. Due of theseto the pigments high commercial in others strains value ofof cyanobacteriathese pigments to Soil samples came from sites established along different in the market, it is suggested to continue with the detection and assess the possibility of their mass culture and exploitation. Keywords: Nostoc commune Bornet & Flahault came from the transect ATA Pigments Content altitudinal transects made° in the Atacama° Desert in July 2009. Atacama Desert; Cryptogamic crusts; Phycobilin Tolypothrix tenuis 11-6B° (north Calama,° 20 56’074”S, 68 57’440”O, 2.964 m) and Introduction Kütz. ex Bornet & Flahault of ATA 2-3 (Vallenar, Cyanobacteria 29Activation22’454’’, 70 and58’936’’O, Incubation 446 m). of the Soil Samples are organisms capable to fix carbon only as that is transformed in ammonium and in this way it is liberated and waterfalls. Previous to the taxonomical determination of higher plants. Also they can fix atmospheric nitrogen, a compound Soil samples were obtained from cryptogamic crusts, pools biofertilizer. They also have phycobilin pigments as: phycocyanin to the soil, enriching it with this element, which acts as natural and phycoerythrin. Cyanobacteria are part of numerous the species, the soil samples of each site of the transects were sieved and then activated in a shaker for 30 min contained 200 ml of distilled water. Aliquots were plated in Petri plates in agar ecosystems both freshwater and marine, but also of desert soil all Z8 and Z8-N media and were grown under controlled conditions: stress.over the Their world. high On deserttolerance ecosystems, to these these conditions microorganisms allows us are to continuous light, room temperature for a period of 3 weeks to a supportcontinuously that these submitted organisms to a strong could hydric,be easy thermic to culture and and nutritive in this month. After that time, the colonies were replicated in Petri plates with agar Z8 and Z8- N media with the purpose to obtain unialgal way achieve the commercialization of these molecules. inocula that afterwards were cultured in 250 ml Erlenmeyer flasks with liquid Z8-N media. Symbiosis Group *Corresponding author email: [email protected] Content of Phycobilin Pigments in Two Strains of Cyanobacteria of the Atacama Desert, Copyright: Chile © 2014 Pereira et al. Taxonomical Determination C-phycocyanin. found in major proportion than the C-phycocyanin. For the determination of the species, morphological and • In both the studied strains, the C-phycoerythrin was Tolypothrix strain had lucidareproductive camera characters and graduated were ocular.used. Measurements The obtained informationwere taken using a Nikon optic microscopy Optiphot model equipped with was submitted to keys and compared with descriptions presented • The Nostoc major quantity of phycobilin in the specialized bibliography such as [3-6]. pigments, with 0.66 mg/ml PE and 0.32 mg/ml of PC. On the other hand, the strain had 0.080 mg/ml PE and Treatment of Samples and Extraction of Phycobilin 0.037 mg/ml PC. Pigments • According to the obtained results, it can be speculated Cyanobacteria ofthat phycocyanin with a biomass can bebetween obtained 100 for and the 200 case mg/ml of Tolypothrix of moist samples had fresh weights, between 70-120 tenuis.weight of algae, 1 mg/ml of phycoerythrin and 0.5 mg/ml mg; 30 mg of sand were added to those samples. After that, they were washed with concentrated HCL, sieved, calcined, dried and market (phycocyanin about of US $535 the milligram macerated in a mortar with fine sand. To homogenize and extract • According to the high value of these pigments in the the pigments, 1 ml of buffer phosphate of pH 6.8 was added to the samples. Then, 2 ml of the treated samples were transferred to an itand is phycoerythrinsuggested to aroundcontinue of withUS $244 the thedetection milligram), and Eppendorf tube and the remnant solution was again washed with determinationdepending on ofthe these type pigments of species in other (Sigma-Aldrich), cyanobacteria two aliquots of 0.5 ml of buffer phosphate. Finally, the samples strains to assess the possibility of their mass culture and Determinationwere centrifuged in of a micro-the Concentration centrifuge at 1000 of rpm Phycobilin for 20 min. exploitation. Pigments The supernatant obtained was measured in a from the more rentable strain (Tolypothrix tenuis) the • To obtain one milligram of phycoerythrin and phycocyanin spectrophotometer at the following wavelength: 455, 565, absorbance were used for the calculation of the phycobilins andcommercial harvest. cost was of US $4 and US $7.5 respectively. pigments592, 618, concentrations645 and 730 nm(phycocyanin using a quartz and phycoerythrin) cell. Readings byof This value includes the cost of reagents, isolation, culture Conclusions three replicates Table 1. the equations of [7]. The results were expressed in an average of Results and Discussion strains of cyanobacteria different Based proportions on the obtained of phyocyanin results, we and can phycoerythrin.conclude that bothThe Both strains presented phycobilin pigments (C-phycoerythrin percentage of yields of thesefrom two the pigments Atacama could Desert be increased possess and C-phycocyanin). Tolypothrix tenuis was the strain that under different types of light allowing to increase the added value presented a major concentration of C- phycoerythrin and cyanobacteria possess. of these natural resources, due to the “chromatic adaptation” that Acknowledgments The authors thank to project 2009-2010 Biodiversity of forterrestrial the facilities cyanobacteria provided inof theAtacama development Desert, of Chile this study.financed by NSF (National Science Foundation) and to the Talca University References 1. Pentón-Rol G, Martínez-Sánchez G, Cervantes-Llanos M et al. Noctoc commune Tolypothrix tenuis C-Phycocyanin ameliorates experimental autoimmune encephalomyelitis Figure 1: Cyanobacteria strains used in obtaining of phycobilin pig- and induces regulatory T cells. Int. Immunopharmacology. 2011; 11: 29- ments. The black scale represents 1 mm. 38. 2. riksen NT. Production of phycocyanin- a pigment with applications in Table 1: Content of phycobilin pigments in the studied strains. biology, biotechnology, foods and medicine. Appl Microbiol Biotechnol. Phycobilin pigments 2008;E 80: -84. C- phycoerythrin C-phycocyanin 3. esikachary TV. Cyanophyta. New Delhi: Indian Council of Agricultural ( (PC) Research. 1959; 1-686. Cyanobacteria Strains/ Content mg/ml mg/ml D PE) Kryptogammenflora Nostoc commune von Deutschland, Osterreich, under

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