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C-phycocyanin extraction from Spirulina platensis wet biomass
Moraes, C. C.;Sala, Luisa;Cerveira, G. P.;Kalil, S. J.;
Brazilian Journal of Chemical Engineering , 2011, DOI: 10.1590/S0104-66322011000100006
Abstract: c-phycocyanin is a natural blue dye used in food and pharmaceutical industry. in the present study, a simple and efficient method to extract c-phycocyanin from spirulina platensis wet biomass is reported. the extractions were carried out using six different methods, including chemical (organic and inorganic acid treatment), physical (freezing and thawing, sonication, homogenization) and enzymatic (lysozyme treatment) methods. the extraction using ultrasonic bath in the presence of glass pearls in the biomass proved to be the most efficient method, 56% higher than using freezing and thawing (the method most frequently used), and presented a extraction yield of 43.75 mg.g-1 and a c-phycocyanin concentration of 0.21 mg.ml-1.
Effects of Chemical Parameters on Spirulina platensis Biomass Production: Optimized Method for Phycocyanin Extraction
P. Soundarapandian,B. Vasanthi
International Journal of Zoological Research , 2008,
Abstract: The micro alga, Spirulina is a rich source of protein, which is used as a protein supplement for humans, chicks and also in aquaculture. Among the cultures, CS-1 registered maximum biomass production and S-20 showed highest biomass production among the local isolates. Optimum temperature of 35C was the best for maximum biomass production of S. platensis cultures. Among the cultures CS-1 culture, put forth maximum biomass production at 35C. The biomass production of all S. platensis cultures was maximum at pH 10.0. Among the cultures, CS-1 recorded maximum biomass at pH 10.0. S. platensis culture S-20 showed highest biomass production among the local isolates. S. platensis cultures were grown under different light wave lengths ranging from 340-700 nm and observed that it grows best in red light but later on there was no significant difference between the biomass produced under red and normal white lights. S. platensis culture CS-1 has shown the highest chlorophyll, carotenoids and phycocyanin and protein contents. When the extracted protein was resolved on a 15% SDS-PAGE gel, the cultures have polypeptide subunits ranging from the molecular weights 20 to 95 kDa. The liquid nitrogen method was found to be the best by extraction higher quantity of phycocyanin from all S. platensis cultures. Among the cultures, S. platensis culture CS-1 recorded the highest phycocyanin content and among the local isolates SM-2 showed the highest pigment content. SDS-PAGE analysis of phycocyanin pigment revealed two characteristic bands with a molecular weights of 14.3 and 20.1 kDa approximately for a and subunits.
Study of phycocyanin production from Spirulina platensis under different light spectra
Walter, Alfredo;Carvalho, Júlio Cesar de;Soccol, Vanete Thomaz;Faria, Ana Bárbara Bisinella de;Ghiggi, Vanessa;Soccol, Carlos Ricardo;
Brazilian Archives of Biology and Technology , 2011, DOI: 10.1590/S1516-89132011000400005
Abstract: the aim of this work was to investigate the production of phycocyanin by spirulina platensis under different spectra of light. the dependent variables evaluated were the amount of phycocyanin obtained and its purity, demonstrating that there might be a restructuring of phycobilisomes, especially when the culture was subjected to red light, which increased the purity level up to 33% with a reduction of 16% in phycocyanin content, but with higher photosynthetic efficiency compared to natural light.
Combinational biosynthesis and characterization of a fluorescent 82β-phycocyanin of Spirulina platensis
XiangYu Guan,WeiJie Zhang,XiaoYuan Chi,HanZhi Lin,JinFeng Wang,Song Qin
Chinese Science Bulletin , 2012, DOI: 10.1007/s11434-012-5264-2
Abstract: To biosynthesize fluorescent Spirulina platensis (Sp) β-phycocyanin (PC) in Escherichia coli, a BLASTP search for homologs of the cpeS gene, a chromophore lyase, was performed against the Synechocystis sp. PCC 6803 (S6) proteome. A highly homologous gene, slr2049, was obtained from the S6 genome. Sites 82 and 153 in β-phycocyanin of Sp were modified by site-directed mutagenesis. Two recombinant expression vectors were constructed and transformed into E. coli BL21: (i) pCDF-cpcB (C153A)-slr2049-sll0583-ho1-pcyA; and (ii) pCDF-cpcB (C82I)-slr2049-sll0583-ho1-pcyA. Lyases encoded by the genes slr2049 and sll0583 catalyzed the linking of Sp 82β-PC to phycocyanobilin (PCB), and fluorescent CpcB (C153A)-PCB was generated. We present a strategy for the co-expression of multiple genes in a single expression vector to identify the function of an unknown gene. Recombinant phycobiliproteins produced on a large scale are promising fluorescent tags for diagnostics and pharmacology.
Experimental design as a tool for optimization of C-phycocyanin purification by precipitation from Spirulina platensis
Silva, Lorena A.;Kuhn, Kátia R.;Moraes, Caroline C.;Burkert, Carlos A. V.;Kalil, Susana J.;
Journal of the Brazilian Chemical Society , 2009, DOI: 10.1590/S0103-50532009000100003
Abstract: c-phycocyanin is a natural blue dye that has been used as an additive in food and can be used to produce medications. the major goal of the present study was to obtain c-phycocyanin under the best operational conditions for high c-phycocyanin recovery and purity using the precipitation technique. crude c-phycocyanin from spirulina platensis was used. optimization of the purification was carried out using experimental design. the effect of ammonium sulfate concentration, volume and of ph for resuspension were evaluated. subsequently an ammonium sulfate fractionation study was carried out using the most suitable conditions found in the experimental design. the best purification condition was ammonium sulfate fractionation at 0-20%/20-50%, in relation to a resuspension volume/initial volume of 0.52 in a 7.0 ph buffer. under these conditions, in an one-step purification only, the purity increased 70% compared to the initial extract, with an 83.8% recovery.
Extraction and purification of C-phycocyanin from Spirulina platensis in conventional and integrated aqueous two-phase systems
Antelo, Francine S.;Anschau, Andréia;Costa, Jorge A. V.;Kalil, Susana J.;
Journal of the Brazilian Chemical Society , 2010, DOI: 10.1590/S0103-50532010000500022
Abstract: crude c-phycocyanin extracts from spirulina platensis, with and without cell debris, were purified in aqueous two-phase systems ( atps) varying the molar mass of the peg, at ph 6, more promising than at ph 7 . for the assays carried out with each molar mass, with distinct percentages of peg and salt, the concentrations (mg ml-1), purities (od 620/od 280) and c-phycocyanin partition coefficients of each system were determined and compared. the process for the primary purification of intracellular c-phycocyanin with cell debris (integrated), composed of 15% of peg 1500 and 13% of potassium phosphate and 5% of peg 4000 and 18% of salt, produced c-phycocyanin concentrations of, respectively, 1.60 and 2.67 mg ml-1 in the top phase. the purities were of 0.73 and 0.79 respectively, values higher than those obtained in the systems without cells.
Purification, crystallization and preliminary crystallographic investigations of selenium-containing phycocyanin from selenium-rich algae (Spirulina platensis)
Lenong Li,Jiping Zhang,Tao Jiang,Baojiang Guo,Wenrui Chang,Dongcai Liang
Science China Life Sciences , 2001, DOI: 10.1007/BF02879600
Abstract: The selenium-containing phycocyanin from the selenium-rich algae (Spirulina platensis) has been crystallized in two crystal forms by the hanging-drop vapor diffusion techniques. A chromatographic procedure of gel filtration and anion exchange was used for purification. Form I crystal with space group P21 and cell parameters a =108.0 , b= 117.0 , c = 184.0 , β= 90.2° and 12(αβ) units in the asymmetric unit was obtained by using (NH4)2SO4 as precipitant. These crystals diffract up to 2.8 . Form II crystal obtained by using PEG4000 as precipitant belongs to space group P63 with unit cell constants a = 155.0 , c = 40.3 , γ =120.0° and one(αβ) unit in the asymmetric unit. The crystals diffract beyond 2.9 . The possible stacking forms of phycocyanin molecules in the first crystal form were discussed.
Purification, crystallization and preliminary crystallographic investigations of selenium-containing phycocyanin from selenium-rich algae (Spirulina platensis)

Lenong Li,Jiping Zhang,Tao Jiang,Baojiang Guo,Wenrui Chang,Dongcai Liang,

中国科学C辑(英文版) , 2001,
Abstract: The selenium-containing phycocyanin from the selenium-rich algae (Spirulina platensis) has been crystallized in two crystal forms by the hanging-drop vapor diffusion techniques. A chromatographic procedure of gel filtration and anion exchange was used for purification. Form I crystal with space group P21 and cell parameters a =108.0 , b= 117.0 , c = 184.0 , β= 90.2° and 12(αβ) units in the asymmetric unit was obtained by using (NH4)2SO4 as precipitant. These crystals diffract up to 2.8 . Form II crystal obtained by using PEG4000 as precipitant belongs to space group P63 with unit cell constants a = 155.0 , c = 40.3 , γ =120.0° and one(αβ) unit in the asymmetric unit. The crystals diffract beyond 2.9 . The possible stacking forms of phycocyanin molecules in the first crystal form were discussed.
New research on extraction and purification of phycocyanin from Spirulina platensis
钝顶螺旋藻藻蓝蛋白提取纯化新工艺

LI Bing,ZHANG Xue-cheng,GAO Mei-hua,ZHU Xian-ming,
李冰
,张学成,高美华,褚现明

海洋科学 , 2007,
Abstract: 对钝顶螺旋藻(Spirulina platensis)中藻蓝蛋白的提取和纯化方法进行了改进。用磷酸盐缓冲液循环冻融联合超声波破碎法,50%硫酸铵沉淀获得藻蓝蛋白(phycocyanin,PC),提取率达到13.1%。粗蛋白提取液再经过两次羟基磷灰石柱(HA)层析和sephacryl-HR-200凝胶层析对其进行纯化,纯度达到4.71%。纯化后的PC在12%十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)中得到两条带,分别是α和β两个亚基,分子质量分别为21.4ku和17.0ku。结果表明,通过上述分离纯化过程得到了较高提取率和电泳纯度的藻蓝蛋白。
Effect of Hg(II) and Pb(II) Ions on C-Phycocyanin (Spirulina platensis)  [PDF]
Eteri Gelagutashvili, Ketevan Tsakadze
Optics and Photonics Journal (OPJ) , 2013, DOI: 10.4236/opj.2013.31020
Abstract:

Influence of Hg(II) and Pb(II) ions on C-Phycocyanin (C-PC) from cyanobacteria Spirulina platensis was investigated using Fluorescence spectroscopy. Fluorescence measurements demonstrate quenching of C-PC emission by Hg(II) and Pb(II), and blue shifts in the fluorescence spectra. The effect of DNA on the fluorescence of Hg(II)-and Pb(II)-C-PC (from Spirulina platensis) complexes was also studied. It was shown that the fluorescence intensity of Hg-C-PC after addition of DNA gave rise to the fluorescence buildup. At the same time, addition of DNA to the Pb(II)-C-PC complexes showed no such effect. In the case of Hg(II)-C-PC, fluorescence intensity significantly decreases in time, while for Pb(II)-C-PC, decrease of the fluorescence intensity is not significant, but blue shift of the peak takes place.

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