Optimization of Spirulina’s Phycocyanin Extraction Yield Using Response Surface Method

Besides cultivation, extraction is also a critical stage in enhancing the yield of phycocyanin production - a highly valuable compound from Spirulina biomass. In this study, the combined effect of three important variables in the ultrasonic-assisted extraction process on phycocyanin extraction yield, namely extraction temperature, sonication time, and solvent pH were investigated through a central composite design experiment. Furthermore, the response surface method was applied in order to define an optimal condition to achieve the highest extraction yield. The results showed that when temperature ranged from 35ºC to 45ºC, sonication time from 20 to 50 minutes, and solvent pH from 6 to 8, the average yield of 30.135±1.552 mg/g was obtained with an average purity of 0.871±0.043. A regression model was also successfully developed, which allowed a good prediction of extraction yield based on the three mentioned variables. On the other hand, an optimal condition for extraction was also proposed with sonication time = 43.57 minutes, extraction temperature = 37.6oC, and solvent pH = 6.7. These results were practically valuable for the improvement of phycocyanin extraction from Spirulina biomass

Effects of Spirulina phycocyanin on antioxidant capacity of ethanol-induced oxidative stress mice / 中国生物制品学杂志

@#Objective To investigate the effect of Spirulina phycocyanin on antioxidant capacity of ethanol-induced oxidative stress mice.Methods Forty-eight female KM mice were selected and randomly divided into four groups:The blank group,model group,low-dose group and high-dose group,12 for each group. Mice in low-dose and high-dose group were given0. 15 g/kg and 0. 30 g/kg phycocyanin by intragastric administration,once a day,continuously for 42 d,and the body mass of mice were weighed. Fasting for 16 h(overnight)after the last intragastric administration,50% ethanol was given once at 12 mL/kg body mass. The mice in model group were only given 50% ethanol by intragastric administration,while the mice in blank group were not given intragastric administration. After 6 h,the blood samples were collected and the sera were separated and detected for the content of 8-isoprostane by corresponding kit. The liver tissues of mice in each group were taken aseptically. After grinding and centrifuging,the supernatant was taken and detected for the contents of protein carbonyl,glutathione peroxidase(GSH-Px)and reduced glutathione(GSH)with corresponding kits. The correlation between mouse body mass and GSH was analyzed.Results Compared with the blank group,the body mass of mice in the model group increased,while the difference was not significant(F = 1. 585,P > 0. 05);the contents of 8-isoprostane in serum,protein carbonyl and GSH in liver tissue significantly increased(F = 11. 697,13. 582 and 17. 213 respectively,each P < 0. 05);the content of GSH-Px in liver tissue decreased,while the difference was not significant(F = 5. 978,P > 0. 05). Compared with the model group,the body mass of mice in low-dose and high-dose groups decreased significantly(F = 4. 125 and 18. 842 respectively,each P < 0. 05);the contents of 8-isoprostane in serum and protein carbonyl in liver tissue decreased significantly(F = 10. 695～40. 512,each P < 0. 01);the contents of GSH-Px and GSH in liver tissue significantly increased(F = 42. 65～76. 379,each P < 0. 01). The content of GSH in liver tissue was negatively correlated with the body mass of mice(R2= 0. 013 49,P > 0. 05).Conclusion Phycocyanin reduced the oxidative damage,improved the antioxidant capacity,and reduced the body mass of ethanol-induced oxidative stress mice.

Extraction and partial characterization of exopolysaccharides and pigments from cyanobacterium Oscillatoria pseudogeminata G.Schmid

Cyanobacterial pigments such as chlorophyll, carotenoids and phycobiliproteins are considered as ecofriendly natural colourants used in food and food additives as they have nutraceutical value, non-toxic and non-carcinogenic. The present study focused on extraction, partial characterization of extracellular polysaccharides (EPS), phycocyanin and mycosporin like amino acid (MAA) from a freshwater cyanobacterium Oscillatoria pseudogeminata G.Schmid.The purified isolate of O.pseudogeminatawas deposited and maintained in the NRMC-F, Bharathidasan University. The EPS was extracted withthe help of double volume of ice-cold acetone and confirmed by UV (260 and 280 nm), FTIR and XRD analyses.Phycocyanin and MAA were also extracted from O. pseudogeminata biomass using double distilled water. The UV-VisSpectrophotometric results revealed that 520 and 230 nm peaks represent phycocyanin and MAA, respectively. Further, theprocess of scaling-up the biomass and increase the productivity of EPS, Phycocyanin and MAA from Oscillatoriapseudogeminata is under raceway pond system, already initialed in our laboratory.

Physicochemical degradation of phycocyanin and means to improve its stability:A short review / 药物分析学报

The cyanobacterium Arthrospira platensis,spirulina,is a source of pigments such as phycobiliprotein and phycocyanin.Phycocyanin is used in the food,cosmetics,and pharmaceutical industries because of its antioxidant,anti-inflammatory,and anticancer properties.The different steps involved in extraction and purification of this protein can alter the final properties.In this review,the stability of phycocyanin(pH,temperature,and light)is discussed,considering the physicochemical parameters of kinetic modeling.The optimal working pH range for phycocyanin is between 5.5 and 6.0 and it remains stable up to 45℃;however,exposure to relatively high temperatures or acidic pH decreases its half-life and increases the degradation kinetic constant.Phycobiliproteins are sensitive to light;preservatives such as mono-and di-saccharides,citric acid,or sodium chloride appear to be effective stabilizing agents.Encapsulation within nano-or micro-structured materials such as nanofibers,microparticles,or nanoparticles,can also pre-serve or enhance its stability.

C-phycocyanin inhibits epithelial-mesenchymal transformation ofTGF-β1-induced cervical cancer Caski cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of C-phycocyanin (C-PC) on the epithelial-mesenchymal transition (EMT) of cervical cancer Caski cells induced by transforming growth factor beta1 (TGF-β1). Methods: According to different treatment methods, Caski cells were divided into three groups: 10 ng/ml TGF-β1 treatment group, 10 ng/ml TGF-β1+300 μg/ml C-PC co-treatment group and control group (untreated). After 24 h of treatment, the morphological changes of Caski cells were observed, and the effects of TGF-β1 and C-PC on the migration and invasion of Caski cells were detected by Scratch test and Transwell test, respectively. Western blotting was used to detect the effect of C-PC on the expression of epithelial phenotypic marker protein E-cadherin and stromal phenotypic marker protein N-cadherin in TGF-β1-induced Caski cells, and qPCR was used to detect the mRNA expressions of EMT related factors Snail, Zeb1 and Twist. Results: Caski cells in the TGF-β1 treatment group lost the characteristics of the original epithelial phenotype, while the cells in the TGF-β1+C-PC co-treatment group maintained the characteristics of normal epithelial phenotype; the migration rate ([60.0±1.4]% vs [33.5±2.2]%, [40.0±2.8]%, both P<0.05) and the number of invasive transmembrane cells ([108.2±6.2] vs [25.2±3.1], [39.8±5.4], both P<0.01]) of Caski cells in the TGF- β1 treatment group were significantly higher than those in the co-treatment group and the control group. Compared with the control group, the expression of E-cadherin in Caski cells treated with TGF-β1 decreased significantly (P<0.05), while the mRNA expressions of Twist, Snail and Zeb1 increased significantly (all P<0.05); However, co-treatment with C-PC reversed above changes (P<0.05 or P<0.01), and significantly decreased the protein expression level of N-cadherin (P< 0.05). Conclusion: C-PC treatment can inhibit the invasion and metastasis ability of Caski cells induced by TGF-β1 and further affects the EMT process. The mechanism may be related to the decrease of mRNAexpressions of Twist, Snail and Zeb1 by C-PC treatment. ·

Targeted antitumor effects of C-PC/CMC-CD55sp nanospheres on cervical carcinoma Caski cells / 中国肿瘤生物治疗杂志

@# Objective: To prepare a new type of phycocyanin/carboxymethyl chitosan-CD55 ligand peptide (CPC/CMC-CD55sp) nanospheres, and to study its targeted therapeutic effect on cervical cancer Caski cells. Methods: The novel CPC/CMC-CD55sp nanospheres (CPC/CMC-CD55sp) were synthesized by ionic cross-linking method, and the properties of nanospheres were observed by transmission electron microscopy (DLS) and fourier transform infrared spectroscopy (FTIR). The expression of CD55 on the surface of Caski and fibroblast (L-929) cells was detected by Western blotting and flow cytometry. The effect of nanospheres on the proliferation of Caski cells was detected by CCK-8. Flow cytometry and fluorescence microscopy were used to detect the uptake of microspheres by Caski cells; Western blotting and flow cytometry were used to detect the effect of CPC/CMC-CD55sp on expressions of apoptosis-related proteins and apoptosis rate in Caski cells; the hemolysis test was used to determine the biological safety of the drug. Results: CPC/ CMC-CD55sp was successfully prepared with good morphology and uniform diameter; and CD55 was highly expressed on the surface of Caski cells but low expressed on the surface of L-929 cells (P<0.01). CPC/CMC-CD55sp could targeted and efficiently reach Caski cells and be ingested into the cells. It exhibited weak hemolysis effect on human peripheral blood, which was in the safe range. CPC/ CMC-CD55sp displayed obvious inhibitory effect on Caski cell proliferation, and could induce cell apoptosis (P<0.05 or P<0.01). Conclusion: The new CPC/CMC-CD55sp can targeted inhibit the growth of cervical cancer Caski cells via inducing its apoptosis and has good bio-safety, which provides a new idea for the research and development of anti-tumor marine drugs.

Principales propiedades inmunomoduladoras y antinflamatorias de la ficobiliproteína C-ficocianina / Main immunomodulatory and anti-inflamatory properties of phycobiliproteins C-phycocyanin

Las ficobiliproteínas son proteínas solubles en agua, que funcionan como pigmentos fotosintéticos accesorios en diferentes organismos tales como las cianobacterias, las algas rojas y las criptomonadas. En el alga verdeazul Spirulina platensis, una de las ficobiliproteínas más abundantes es la C-ficocianina, la cual tiene unido tres cromóforos ficocianobilina mediante un enlace tioéter a cisteínas específicas. La ficocianobilina es un tetrapirrol lineal asociado a la captación de energía solar en estos organismos. La C-ficocianina ha sido empleada en diferentes investigaciones biomédicas como biomarcador, por sus propiedades fluorescentes, y como posible agente terapéutico para el tratamiento de enfermedades asociadas al estrés oxidativo, por sus propiedades antioxidantes, inmunomoduladoras y antinflamatorias. Se ha demostrado que esta proteína aumenta la liberación de interferón gamma en células mononucleares de sangre periférica y modula la producción de citocinas inflamatorias como el factor de necrosis tumoral alfa, entre otras. Además, se ha encontrado que la C-ficocianina tiene efecto inmunomodulador de citocinas que potencian la activación de las células del sistema inmune, como la IL-6 y la IL-1ß, así como la regulación de aproximadamente 190 genes implicados en la inmunidad(AU)

Phycobiliproteins are water-soluble proteins that function as accessory photosynthetic pigments in different organisms such as cyanobacteria, red algae and cryptomonads. In the blue-green algae Spirulina platensis one of the most abundant phycobiliproteins is the C-phycocyanin, which has three phycocyanobilin chromophores linked through a thioether bond to specific cysteine. The phycocyanobilin is a linear tetrapyrrole associated with solar energy absorption in these organisms. The C-phycocyanin has been used in several biomedical researches as a biomarker, for their fluorescence properties, and as a possible therapeutic agent for the treatment of diseases associated with oxidative stress for its antioxidant, anti-inflammatory and immunomodulatory properties. It has been shown that this protein increases the release of interferon gamma in peripheral blood mononuclear cells, and modulates the production of inflammatory cytokines such as tumor necrosis factor among others. Furthermore it has been found that the C-phycocyanin has immunomodulatory effect on cytokines that enhance the activation of immune cells, such as IL-6 and IL-1ß, and the regulation of about 190 genes involved in immunity(AU)

Role of pH on antioxidants production by Spirulina (Arthrospira) platensis

Abstract Algae can tolerate a broad range of growing conditions but extreme conditions may lead to the generation of highly dangerous reactive oxygen species (ROS), which may cause the deterioration of cell metabolism and damage cellular components. The antioxidants produced by algae alleviate the harmful effects of ROS. While the enhancement of antioxidant production in blue green algae under stress has been reported, the antioxidant response to changes in pH levels requires further investigation. This study presents the effect of pH changes on the antioxidant activity and productivity of the blue green alga Spirulina (Arthrospira) platensis. The algal dry weight (DW) was greatly enhanced at pH 9.0. The highest content of chlorophyll a and carotenoids (10.6 and 2.4 mg/g DW, respectively) was recorded at pH 8.5. The highest phenolic content (12.1 mg gallic acid equivalent (GAE)/g DW) was recorded at pH 9.5. The maximum production of total phycobiliprotein (159 mg/g DW) was obtained at pH 9.0. The antioxidant activities of radical scavenging activity, reducing power and chelating activity were highest at pH 9.0 with an increase of 567, 250 and 206% compared to the positive control, respectively. Variation in the activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) was also reported. While the high alkaline pH may favor the overproduction of antioxidants, normal cell metabolism and membrane function is unaffected, as shown by growth and chlorophyll content, which suggests that these conditions are suitable for further studies on the harvest of antioxidants from S. platensis.

Inhibition of Fibrotic Contraction by C-Phycocyanin through Modulation of Connective Tissue Growth Factor and α-Smooth Muscle Actin Expression

The effects of C-phycocyanin (C-pc), a phycobiliprotein, on the expression of pro-fibrotic mediators in hyper-tropic scarring such as connective tissue growth factor (CTGF) and α-smooth muscle actins (α-SMA) were investigated in relation to trans-differentiation of fibroblast to myo-fibroblast, an icon of scar formation. C-pc was isolated from Spirulina Platensis extract using sonication method and C-pc concentration was determined by Bennet and Bogorad equation. α-SMA and CTGF levels in wounded primary human dermal fibroblasts were determined by western blot analysis and immuno-fluorescence confocal microscope was employed. Fibroblast contractility was examined by three-dimensional collagen lattice contraction assay. There was an elevation of α-SMA (121%) and CTGF (143%) levels in wound cells as compared with non-wound cells. The does-response profiles of down regulation demonstrated that the maximum inhibitions of α-SMA by 63% (p<0.05) and CTGF by 50% (p<0.1) were achieved by C-pc (6 nM) treated cells. In confocal assay, non-wound fibroblasts exhibited basal level of α-SMA staining, while wounded cells without C-pc treatment showed strong up-regulation of α-SMA by 147% (p<0.05). C-pc (6 nM) inhibited α-SMA expression by 70% (p<0.05) and reduced collagen contraction by 29% (p<0.05). C-pc seemed to lessen the over expression of CTGF, α-SMA, subsequently alleviating the fibrotic contracture. This study suggests the potential application of C-pc to regulation of the expression of pro-fibrotic mediators in scarring process and its potential usage as an efficient means for anti-fibrosis therapy.

Purification of C-phycocyanin from Spirulina platensis in aqueous two-phase systems using an experimental design

C-phycocyanin from Spirulina platensis was purified in aqueous two-phase systems (ATPS) of polyethylene glycol (PEG)/potassium phosphate, varying the molar mass of the PEG. Results using a full factorial design showed that an increase in the concentration of salt and decrease in the concentration of PEG caused an increment in the purification factor for all the ATPS studied. Optimization of the conditions of the purification was studied using a central composite rotatable design for each molar mass of PEG. The ATPS composed of 7% (w/w) PEG 1500 or 4% (w/w) PEG 8000 (g/gmol) and 23 or 22.5% (w/w) of phosphate resulted a purification factor of 1.6-fold for C-phycocyanin, with total and 57% recovery, respectively. Process conditions were optimized for the purification factor for the system with PEG 1500. The ATPS with 4% (w/w) PEG 4000 or 4% (w/w) PEG 6000 and 21% (w/w) phosphate resulted purification factors of 2.1 and 2.2-fold, recovering 100% and 73.5%, respectively of C-phycocyanin in the top phase.

Effects of phycocyanin on apoptosis of human laryngeal cancer HEP-2 cells / 中国病理生理杂志

[ ABSTRACT] AIM:To investigate the effect of phycocyanin on the apoptosis of human laryngeal cancer HEP-2 cells and to explore the inhibitory mechanism of phycocyanin to tumor.METHODS:Highly purified phycocyanin was ex-tracted from spirulina.The effects of phycocyanin at different concentrations on the growth of human laryngeal cancer HEP-2 cells were detected by MTT assay.In addition, the cell structures were observed under electron microscope.The cell ap-optosis was analyzed by flow cytometry.The production of reactive oxygen species ( ROS) was measured by flow cytometry. Enzymatic activities of caspase-3,-8 and-9 were measured by chemical colorimatry.The expression of Bax, Bcl-2, Fas, P53, caspase-3 and caspase-9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS:MTT test confirmed that phycocyanin inhibited the cell activity of HEP-2 cells with time and dose dependent manners.The result of electron microscope observation and flow cytometry indicated that phycocyanin induced the apoptosis of HEP-2 cells.The intracellular content of ROS was increased.The activities of caspase-3, -8 and -9 were increased.RT-PCR showed that the mRNA expression of Bax, Fas, P53, caspase-3, caspase-9 was increased and Bcl-2 was decreased.The results of Western blot were consistent with the results of RT-PCR.CONCLUSION:Phycocyanin might induce apoptosis of HEP-2 cells by down-regulating Bcl-2, up-regulating Bax, Fas and P53, and the transduction of apoptotic signals in the human laryngeal cancer cells.

C-phycocyanin protects doxorubicin-induced myocardial cells injury by Nrf 2/heme oxygenase-1 pathway / 中国生化药物杂志

Objective To observe the role of heme oxygenase (HO)-1 on the protective effect of C-phycocyanin (CPC) on doxorubicin (DOX)-induced myocardial cells injury by Nvf2/HO-1 pathway. Methods 60 SD rats were randomly divided into control group, DOX group, CPC group and tin protoporphyrin IX (SnPP, an inhibitor of HO-1) group. The control group was injected with normal saline injection,while the DOX group was administrated with doxorubicin by intraperitoneal injection in a cumulative dose of 15 mg/kg for two weeks. For the CPC rats, 20, 40 and 60 mg/kg of CPC was administrated. The level of creatine kinase (CK) and lactate dehydrogenase (LDH) were detected, and the activity of HO-1 and caspase-3 were also examined. Expression of HO-1 and activation of Nrf 2 were detected by Western blot. Results Compared with control group, serum levels of CK, LDH and Caspase-3 activity in DOX group were significantly increased(P<0.05), but HO-1 in cardiac muscle was only increased slightly. upregulation. Treatment with CPC could significantly ameliorated the CK, LDH and Caspase-3 activity, and markedly induce HO-1 expression and its activity. The reduction of CK, LDH and Caspase-3 activity by CPC could be reversed by treatment of the HO-1 inhibitor, SnPP. Furthermore, CPC sould also induce Nrf 2 activation. Conclusion The protective effect of CPC on doxorubicin-induced myocardial cells in jury via Nrf 2 induced HO-1 HO-1 expression.

C-phycocyanin induces Heme oxygenase-1 expression to protect acute lung injury in septic rats / 中国生化药物杂志

Objective To observe the protective effect and molecular mechanism of C-phycocyanin (CPC) on acute lung injury (ALI) in septic rats. Method 75 SD rats were randomly divided into control group, model group and CPC group. Cecal ligation and puncture was used to establish a septic acute lung injury rats (model group). For the CPC groups, septic acute lung injury rats were administrated by 20, 40 and 60 mg/kg CPC by intraperitoneal injection. 72 h after the operation, serum and lung tissue were obtained, the wet to dry weight ratio, the content of TNF-α、IL-6 and IL-10 in bronchoalveolar lavage fluid, the activity of myeloperoxidase (MPO) was analyzed. Expression of heme oxygenase (HO)-1,activation of nuclear factor erythroid 2-related factor 2 (Nrf 2) and nuclear factor-kappa B (NF-B) were detected by Western blot. Superoxide and Nitrite/Nitrate Level production in Lungs and bronchoalveolar lavage fluid were measured by chemiluminescence and reduction method, respectively. Results Treatment with CPC significantly inhibited septic-induced inflammatory responses including elevation of superoxide formation, myeloperoxidase activity, leucocytes and protein infiltration in lung tissues, and production of proinflammatory cytokine, and nitrite/nitrate in bronchoalveolar lavage fluid (P<0.05). In addition, CPC could activate Nrf 2 and induce HO-1 expression, and inhibit NF-B activation in ALI rats. However, blocking HO-1 activity by tin protoporphyrin IX (SnPP), an HO-1 inhibitor, markedly abolished these beneficial effects of CPC in septic-induced ALI. Conclusion The protection mechanism of CPC may be through HO-1 induction and suppressing of NF-kB-mediated inflammatory responses.

Cultivos discontinuos alimentados con urea de la cianobacteria Phormidium sp. en función de la salinidad y edad del cultivo / Urea fed-batch cultures of the cyanobacterium Phormidium sp. as a function of the salinity and age of cultures

Se comparó la eficiencia de sistemas de cultivos discontinuos alimentados versus cultivos discontinuos convencionales, en cuanto a concentración de nitrógeno, adicionando 0,2 mM de urea cada tres días al final de la fase exponencial, durante 21 días. Se realizaron cultivos con un volumen de 1500 mL a 15 y 35 UPS de salinidad, enriquecidos con medio ALGAL 8mM NaNO3, a 238 µmol q m-2 s-1, aireación constante, fotoperiodo 12:12 horas y temperatura de 29 ±3°C. Phormidium sp. posee la capacidad de hidrolizar la urea; mostrando una asimilación de 65±7,07% de la misma, con la mayor producción (p<0,05) de clorofila a, ficocianina y proteínas de 20,26±1,24; 203,47±12,83 y 707,87±28,47 µg mL-1en los cultivos alimentados. La producción de pigmentos vario en el tiempo, independientemente a la salinidad y sistema de cultivo, mientras que la producción de proteínas y carbohidratos totales fue directamente proporcional a la edad del cultivo, con valores máximos de 612,74 ± 5,41 µg mL-1 y 8,96±0,08 mg mL-1 respectivamente a los 31 días. La síntesis de lípidos y EPS fueron influenciadas (p<0,05) por la salinidad, presentando los máximos de lípidos a 15 UPS con 12,22±2,91µg mL-1, y los EPS se incrementaron a 35 UPS con 2,00 ± 0,26 y 2,03 ± 0,15 mg mL-1. Estos resultados determinan que los cultivos de Phormidium sp. alimentados con urea y a salinidades de 15 y 35 UPS, representan una alternativa económica para la producción de clorofila a, ficocianina y proteínas, incrementándose un 31,04; 40,72 y 31,94 % respectivamente en comparación con cultivos no alimentados.

Fed-batch system efficiency versus batch cultures was compared in relation to nitrogen concentration, adding 0,2mM urea at the end of the exponential phase, during 21 days. Cultures were carried out in 1500 mL to 1.5 and 3.5 UPS of salinity, enriched with Algal medium 8mM NaNO3, 238 mol q m-2 s-1, constant aeration, photoperiod 12:12 h. and 29 ±3°C. Phormidium sp. is able to hydrolyze urea; showing a total assimilation of 65±7.07%, with the highest (p< 0.05) chlorophyll a, phycocyanin and protein production of 20.26 ± 1.24, 203.47 ± 12.83 and 707.87 ± 28.47 µg mL-1 in the fed-batch cultures. On the other hand, pigment production varies in time, regardless salinity and culture system. Proteins and total carbohydrate production were directly proportional to the age of cultures, with maximum values of 612.74 ± 5.41 µg mL-1 and 8.96 ± 0.08 mg mL-1, respectively. Lipid and EPS were influenced (p< 0.05) by salinity, showing maximum of lipids at 15 UPS with 12.22±2.91 µg mL-1, and EPS at 15 and 35 UPS with 2.00 ± 0.26 and 2.03 ± 0.15 mg mL-1. These results determine that Phormidium sp. cultures fed with urea, to salinities of 15-35 UPS, represent an economic alternative for chlorophyll a, phycocyanin and protein production, with an increase of 31.04, 40.72 and 31.94% respectively in comparison with non-fed cultures.

Statistical evaluation of nutritional components impacting phycocyanin production in Synechocystis SP

Alkaliphilic cyanobacterial cultures were isolated from Lonar lake (MS, India). Among the set of cultures, Synechocystis sp, was studied for phycocyanin production. A maximum yield was obtained in BG-11 medium at optimized conditions (pH 10 and 16 h light). In order to increase the phycocyanin yield media optimization based on the eight media components a Plackett-Burman design of the 12 experimental trials was used. As per the analysis CaCl2.2H2O and Na2CO3 have been found to be the most influencing media components at 95% significance. Further the optimum concentrations of these components were estimated following a Box Wilson Central Composite Design (CCD) with four star points and five replicates at the center points for each of two factors was adopted for optimization of these two media components. The results indicated that there was an interlinked influence of CaCl2.2H2O and Na2CO3 on 98% significance. The maximum yield of phycocyanin (12% of dry wt) could be obtained at 0.058 g/l and 0.115 g/l of CaCl2.2H2O and Na2CO3, respectively.

Novel substrate (algal protein) for cultivation of Rhodospirillum rubrum.

Rhodospirillum rubrum was grown under light anaerobic conditions with phycocyanin (C-pc) extracted from Spirulina platensis as the sole source of carbon and nitrogen. When grown under these conditions cellular components like lipids, carbohydrates, protein, carotenoids, bacteriochlorophyll were similar to the one grown with malic acid and ammonium chloride. Growth of R. rubrum increased with increase in concentration of C-pc (200 to 1000 mg/l). R. rubrum also utilized C-pc under dark anaerobic condition. With both malic acid and C-pc as carbon sources C-pc was consumed only after exhaustion of malic acid under light anaerobic condition. No aberration of cell morphology was seen under scanning electron microscope (SEM). R. rubrum utilized both phycocyanobilin and phycoprotein individually as well as in combination. When grown with 1000 mg/l of phycoprotein 450 mg/l of biomass was obtained, and with combination of phycocyanobilin (75 mg/l) and phycoprotein (925 mg/l) 610 mg/l of biomass was obtained. Phycocyanobilin alone did not inhibit the growth of R. rubrum. Utilization of C-pc with protease like activity was observed in plate assay. Protease like activity was also observed as zones around the colonies in plates containing sterilized casein, gelatin and filter sterilized bovine serum albumin. No amino acids were detected in the supernatant when analyzed with ninhydrin. Extracellular protease like activity was highest when C-pc was used as substrate (2.8 U/ml). Intracellular protease like activity was not detected in cell free extracts.

Study of phycocyanin production from Spirulina platensis under different light spectra

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 percent with a reduction of 16 percent in phycocyanin content, but with higher photosynthetic efficiency compared to natural light.

Antioxidant effect of phycocyanin on oxidative stress induced with monosodium glutamate in rats

The objective of this work was to study the antioxidant effect of phycocyanin on the oxidative stress induced by monosodium glutamate in the rats. The tests were performed with 32 rats of Wistar breed, divided into four groups, which were administered saline solution of phycocyanin, monosodium glutamate and monosodium glutamate plus phycocyanin. Sulfhydryl groups and the secondary substances derived from lipid oxidation were determined through the level of TBA. The evaluation of these values and the level of sulfhydryl showed that the administration of phycocyanin presented significant antioxidant effect (p < 0.05) reducing the oxidative stress induced by the monosodium glutamate in vivo.

Cysteine rich cyanopeptide β2 from Spirulina fussiformis exhibits plasmid DNA pBR322 scission prevention and cellular antioxidant activity.

Isolation of three different active peptides from C-phycocyanin (C-pc) β chain of S. fussiformis and their biological properties are reported. Phycocyanin peptide β fraction 2 (cyanopeptide β 2) facilitated both antioxidant and plasmid DNA strand scission prevention activity due to higher cysteine moieties in the isolated peptide. The peptide significantly scavenged the free radicals like 1-1,-diphenyl-2-picryl hydrazyl and ferric reducing ability of plasma, increased the absorbance values in reducing power and also showed the higher trolox equivalent antioxidant capacity values in total reactive antioxidant potentials assay. Cyanopeptide β 2 also inhibited reactive oxygen species induced DNA pBR322 damage in dose dependent manner along with free radical scavenging properties suggesting the role in the DNA integrity which is also evident by DNA binding activity of peptide. In addition, the generation of reactive oxygen species (ROS) was dose dependent (10 and 20 ng/ml) and significantly quenched by cyanopeptide β2 in human fibroblast cell line TIG 3-20. In vitro cell scratch injury assay demonstrated the capacity of cyanopeptide β2 in cell migration in to wounded area suggesting fibroblast proliferation and migration. The results suggest that cyanopeptide β2 can be a free radical scavenger and effective peptide for future biomedical applications like wound healing, atherosclerosis, cell redox potential and hypoxia.

C-phycocyanin Attenuates Renal Inflammation and Fibrosis in UUO Mice / 대한신장학회지

PURPOSE: It has been reported that Spirulina, a blue-green algae with potent antioxidant properties, affords significant protection against inflammation and fibrosis in the liver in vivo. The aim of the present study was to establish the possible protective role of C-phycocyanin, one of the active ingredients of Spirulina, in an experimental model of fibrosis in the kidney. METHODS: The study was carried out using male C57BL6 mice. Mice were divided into the following four groups: sham-operated group; C-phycocyanin (PC)-treated sham group; unilateral ureteral obstruction (UUO) group; and PC with UUO group. We evaluated renal TGF-beta mRNA, MCP-1, and osteopontin using real-time RT PCR. We evaluated renal TGF-beta, alpha-SMA, and CD68 by immunohistochemistry. We recorded light microscopic findings of kidney specimens. RESULTS: PC significantly decreased the expression of MCP-1 and alpha-SMA mRNA. Renal gene levels of expression of TGF-beta, MCP-1, and osteopontin in the UUO group were significantly higher than the sham-operated group (p<0.01). The levels of expression of TGF-beta, MCP-1, and osteopontin mRNA of kidneys in the PC-treated UUO group were significantly lower than the untreated UUO group (p< 0.05). The magnitude of expression of TGF-beta and alpha-SMA protein in the kidneys of the PC-treated UUO group was significantly less than the untreated UUO control group (p<0.05). CONCLUSION: The results of the present study suggest that PC has anti-inflammatory and anti-fibrotic effects in an experimental UUO murine model.