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1.
Mar Drugs ; 22(6)2024 May 28.
Artículo en Inglés | MEDLINE | ID: mdl-38921557

RESUMEN

Cyanobacterial phycocyanin pigment is widely utilized for its properties in various industries, including food, cosmetics, and pharmaceuticals. Despite its potential, challenges exist, such as extraction methods impacting yield, stability, and purity. This study investigates the impact of the number of freeze-thaw (FT) cycles on the extraction of phycocyanin from the wet biomass of four cyanobacteria species (Arthrospira platensis, Chlorogloeopsis fritschii, Phormidium sp., and Synechocystis sp.), along with the impact of five extraction solutions (Tris-HCl buffer, phosphate buffer, CaCl2, deionized water, and tap water) at various pH values. Synechocystis sp. exhibited the highest phycocyanin content among the studied species. For A. platensis, Tris-HCl buffer yielded maximum phycocyanin concentration from the first FT cycle, while phosphate buffer provided satisfactory results from the second cycle. Similarly, Tris-HCl buffer showed promising results for C. fritschii (68.5% of the maximum from the first cycle), with the highest concentration (~12% w/w) achieved during the seventh cycle, using phosphate buffer. Phormidium sp. yielded the maximum pigment concentration from the first cycle using tap water. Among species-specific optimal extraction solutions, Tris-HCl buffer demonstrated sufficient extraction efficacy for all species, from the first cycle. This study represents an initial step toward establishing a universal extraction method for phycocyanin from diverse cyanobacteria species.


Asunto(s)
Biomasa , Cianobacterias , Ficocianina , Solventes , Ficocianina/aislamiento & purificación , Ficocianina/química , Cianobacterias/química , Solventes/química , Congelación , Concentración de Iones de Hidrógeno
2.
Arch Microbiol ; 206(6): 258, 2024 May 12.
Artículo en Inglés | MEDLINE | ID: mdl-38735006

RESUMEN

Phycocyanin, a blue-coloured pigment, predominantly found and derived from Spirulina sp., has gained researchers' interest due to its vibrant hues and other attractive properties like antioxidant and anti-microbial. However, the lack of reliable and sustainable phycocyanin extraction strategies without compromising the quality has hindered the scaling up of its production processes for commercial purposes. Here in this study, phycocyanin was extracted from wet and dry biomass Spirulina sp., using three different physical cell disruption methods (ultrasonication, homogenization, and freeze-thaw cycles) combined with two different buffers (phosphate buffer and acetate buffer) and water (as control). The result showed that the freeze-thaw method combined with acetate buffer produced the highest yield (25.013 ± 2.572 mg/100 mg) with a purity ratio of 0.806 ± 0.079. Furthermore, when subjected to 30% w/v salt stress, 1.9 times higher phycocyanin yield with a purity ratio of 1.402 ± 0.609 was achieved using the previously optimized extraction method.


Asunto(s)
Ficocianina , Estrés Salino , Spirulina , Ficocianina/metabolismo , Ficocianina/aislamiento & purificación , Spirulina/metabolismo , Spirulina/química , Biomasa , Congelación
3.
BMC Biotechnol ; 21(1): 40, 2021 06 16.
Artículo en Inglés | MEDLINE | ID: mdl-34134665

RESUMEN

BACKGROUND: Most commercial phycocyanins are extracted from a filamentous cyanobacterium, Arthrospira (Spirulina) platensis. Owing to the expenses of culture and complexities of the physical and chemical methods of phycocyanin purification, a more effective and simple method is required. RESULTS: We developed a new method for efficiently recovering the blue pigment protein, phycocyanin, from unique filamentous cyanobacteria, Pseudanabaena sp. ABRG5-3 and Limnothrix sp. SK1-2-1. The cells were cultivated in economy medium BG11 and lysed by adding water in a 1:16 ratio of wet cells to water. After extraction and purification, 28-30% dry cell weight of phycocyanin was obtained and its purity was confirmed. The stabilities of the phycocyanins at different pH in the presence of high temperature and light conditions and their antioxidant abilities were assessed. Results indicated that the phycocyanins were stable and possessed antioxidant properties. Interestingly, the Pseudanabaena phycocyanin was less likely to deteriorate under acidic conditions. CONCLUSIONS: Overall, we developed a promising and novel method for producing high functional phycocyanin concentrations at a low cost. The possibilities of adapting this new phycocyanin biorefinery to unique bioreactor utilization have also been discussed.


Asunto(s)
Antioxidantes/aislamiento & purificación , Fraccionamiento Químico/métodos , Ficocianina/química , Ficocianina/aislamiento & purificación , Spirulina/química , Antioxidantes/química , Calor , Concentración de Iones de Hidrógeno , Ficocianina/metabolismo , Spirulina/metabolismo
4.
Chem Phys Lipids ; 236: 105064, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33609502

RESUMEN

This study describes the physicochemical properties of soybean asolectin (ASO) liposomes loaded with phycocyanin (Phy) extracted from Spirulina sp. LEB 18. The effects of Phy in the liposomes' properties were investigated by Fourier transform infrared spectroscopy (FTIR), 1H and 31P nuclear magnetic resonance (NMR), zeta (ζ)-potential, dynamic light scattering (DLS) and ultraviolet-visible (UV-vis) techniques. Phy restricted the motion of ASO polar and interface groups and disrupted the package arrangement of the lipid hydrophobic regions, as a likely effect of dipolar and π interactions related to its amino acid residues and pyrrole portions. These interactions were correlated to antiradical/antioxidant Phy responses obtained by 2,2-diphenyl-1-picrylhidrazil (DPPH) assay, thiobarbituric acid reactive substances (TBARS) and ferric reducing antioxidant power (FRAP) methods, and discussed to bring new chemical perspectives about Phy-loaded liposomes-related nutraceutical applications in inflammatory and viral infection processes.


Asunto(s)
Antioxidantes/farmacología , Ficocianina/farmacología , Spirulina/química , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Compuestos de Bifenilo/antagonistas & inhibidores , Recuperación de Fluorescencia tras Fotoblanqueo , Liposomas/química , Ficocianina/química , Ficocianina/aislamiento & purificación , Picratos/antagonistas & inhibidores
5.
Plant Cell Physiol ; 62(2): 334-347, 2021 May 11.
Artículo en Inglés | MEDLINE | ID: mdl-33386854

RESUMEN

Linear tetrapyrrole compounds (bilins) are chromophores of the phytochrome and cyanobacteriochrome classes of photosensors and light-harvesting phycobiliproteins. Various spectroscopic techniques, such as resonance Raman, Fourier transform-infrared and nuclear magnetic resonance, have been used to elucidate the structures underlying their remarkable spectral diversity, in which the signals are experimentally assigned to specific structures using isotopically labeled bilin. However, current methods for isotopic labeling of bilins require specialized expertise, time-consuming procedures and/or expensive reagents. To address these shortcomings, we established a method for pressurized liquid extraction of phycocyanobilin (PCB) from the phycobiliprotein powder Lina Blue and also the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis). PCB was efficiently cleaved in ethanol with three extractions (5 min each) under nitrogen at 125�C and 100 bars. A prewash at 75�C was effective for removing cellular pigments of Synechocystis without PCB cleavage. Liquid chromatography and mass spectrometry suggested that PCB was cleaved in the C3-E (majority) and C3-Z (partial) configurations. 15N- and 13C/15N-labeled PCBs were prepared from Synechocystis cells grown with NaH13CO3 and/or Na15NO3, the concentrations of which were optimized based on cell growth and pigmentation. Extracted PCB was reconstituted with a recombinant apoprotein of the cyanobacteriochrome-class photosensor RcaE. Yield of the photoactive holoprotein was improved by optimization of the expression conditions and cell disruption in the presence of Tween 20. Our method can be applied for the isotopic labeling of other PCB-binding proteins and for the commercial production of non-labeled PCB for food, cosmetic and medical applications.


Asunto(s)
Cianobacterias/metabolismo , Marcaje Isotópico/métodos , Ficobilinas/aislamiento & purificación , Ficocianina/aislamiento & purificación , Fitocromo/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Ficobilinas/química , Ficocianina/química , Synechocystis/metabolismo , Temperatura
6.
FEBS Open Bio ; 11(1): 164-172, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-33190413

RESUMEN

Cyanophyta-phycocyanin (C-PC) is the main constituent of the rod of phycobilisome (PBS), which is a highly ordered and large peripheral light-harvesting protein complex present on the cytoplasmic side of the thylakoid membrane in cyanobacteria and red algae. The C-PC monomer comprises two chains, α- and ß-subunits, and aggregates to form ring-shaped trimers (αß)3 with rotational symmetry. The ring-shaped trimer (αß)3 is a structural block unit (SBU) that forms the rod of PBS. Two (αß)3 SBUs are arranged in a face-to-face manner to form an (αß)6 -hexamer. In this study, the electronic states of three phycocyanobilins, α84, ß84, and ß155 in C-phycocyanin, constituting the rod of the PBS, were calculated for both the trimer and hexamer models by considering the effect of the electrostatic field of protein moieties and water molecules. For the hexamer, the absorption wavelengths of α84, ß84, and ß155 were similar to those obtained experimentally; however, for the trimer, only the absorption wavelength of ß155 shifted toward a shorter-wavelength. The nature of the hexamer structure as a hierarchical structure is revealed by considering the calculated absorption wavelength and energy transfer.


Asunto(s)
Cianobacterias/fisiología , Ficocianina/ultraestructura , Estructura Cuaternaria de Proteína/fisiología , Rhodophyta/fisiología , Cristalografía por Rayos X , Modelos Moleculares , Ficocianina/aislamiento & purificación , Ficocianina/metabolismo , Multimerización de Proteína/fisiología , Relación Estructura-Actividad
7.
J Biosci Bioeng ; 131(2): 161-167, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33109478

RESUMEN

C-Phycocyanin (PC) is a protein used commercially as a natural blue pigment produced by cyanobacteria, cryptophytes, and rhodophytes. Although it is industrially synthesized from the cyanobacterium Arthrospira platensis, PC requires high levels of energy for its extraction, which involves freezing of cells. However, as a protein, PC is easily denatured at extreme temperatures. In this study, we extracted PC from the red alga Cyanidioschyzon merolae, denoted as CmPC, and found that this protein was tolerant to high temperatures and acidic pH. CmPC was extracted by suspending cells in water mixed with various salts and organic acids without freeze-drying or freeze-thaw. The stability of CmPC varied with salt concentration and was destabilized by organic acids. Our results indicate that C. merolae is a potential candidate for PC production with thermotolerant properties.


Asunto(s)
Ficocianina/aislamiento & purificación , Ficocianina/metabolismo , Rhodophyta/enzimología , Rhodophyta/fisiología , Termotolerancia , Calor , Concentración de Iones de Hidrógeno
8.
Int J Biol Macromol ; 165(Pt A): 1111-1118, 2020 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-33031854

RESUMEN

An overview of the biological properties of phycocyanin (PC) amply illustrates that it may not have any specific functional feature towards any system at which it may elicit a specific function, but for the molecular interactions. Nevertheless, based on existing evidences, it is hypothesized that PC has more than one functional target with the interacting systems; therefore, it has diversity of effects. The mechanism of PC action remains elusive of a comprehensive idea. The present investigation focuses on the pro inflammatory enzyme, lipoxygenase (LOX) inhibiting property of PC purified from Oscillatoria sp. Enzyme kinetics studies show that the molecular composite of PC is required for its inhibition shown on LOX. Isothermal titration calorimetric study proves that one molecule of PC interacts with two molecules of LOX. Molecular dynamics simulation study pertaining to PC-LOX interactions shows it to be appropriate as a model to give molecular mechanistic insight into the varied biological properties of PC, demonstrated elsewhere in experimental studies including animal model studies. It explains that the PC-LOX interaction is of a function-freezing, protein-protein interaction in nature. The wide spectrum of properties of PC might be due to its function as a powerful protein hub showing non-specific protein-protein interactions.


Asunto(s)
Inhibidores de la Lipooxigenasa/química , Lipooxigenasa/química , Oscillatoria/química , Ficocianina/química , Animales , Calorimetría , Dominio Catalítico/efectos de los fármacos , Humanos , Lipooxigenasa/efectos de los fármacos , Inhibidores de la Lipooxigenasa/farmacología , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Ficocianina/aislamiento & purificación , Ficocianina/farmacología , Unión Proteica/efectos de los fármacos , Mapas de Interacción de Proteínas/efectos de los fármacos
9.
Biomed Pharmacother ; 132: 110826, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33068929

RESUMEN

OBJECTIVE: Radiation pneumonia and fibrosis are major clinical complications of radiotherapy for thoracic tumor patients, and may significantly reduce survival and quality of life. At present, no safe and effective radiation protection measures have been approved for clinical use. Phycocyanin, a protein responsible for photosynthesis from Spirulina, has been shown to have a variety of biological activities and to be beneficial for a variety of diseases, including pulmonary fibrosis. However, the preventive and protective effects of phycocyanin on radiation-induced pulmonary fibrosis have not been studied. DESIGN: X-ray single dose irradiation was used on the chest of mice to prepare a mouse model of pulmonary fibrosis, from which the effect of phycocyanin on pulmonary histopathologic change, pulmonary fibrosis, the microbiota in lung and gut, LPS, TNF-α, and IL-6 at different time after irradiation were evaluated. RESULTS: Phycocyanin alleviated the radiation-induced lung injury and reduced the level of inflammatory factors. Thorax irradiation led to the disorder in microbiota of the lung and gut. The variation trend of the diversity of the two tissues was opposite, but that of the microbiota composition was similar. The phycocyanin intervention regulated the composition of the lung and gut microbiota, transformed them into normal state, and reduced the level of LPS, which significantly reduced the abundance of inflammation-related bacteria, and increased the abundance of probiotics that produce short-chain fatty acids. CONCLUSION: Phycocyanin could regulate the radiation-induced disorder in lung and gut microbiota of mice, and reduce the radiation-induced lung inflammation and fibrosis.


Asunto(s)
Microbioma Gastrointestinal/efectos de los fármacos , Ficocianina/farmacología , Fibrosis Pulmonar/prevención & control , Neumonitis por Radiación/prevención & control , Animales , Microbioma Gastrointestinal/efectos de la radiación , Inflamación/tratamiento farmacológico , Inflamación/etiología , Inflamación/microbiología , Lipopolisacáridos , Masculino , Ratones , Ratones Endogámicos C57BL , Ficocianina/aislamiento & purificación , Fibrosis Pulmonar/etiología , Fibrosis Pulmonar/microbiología , Traumatismos Experimentales por Radiación/microbiología , Traumatismos Experimentales por Radiación/prevención & control , Neumonitis por Radiación/microbiología , Spirulina/química
10.
Molecules ; 25(15)2020 Jul 28.
Artículo en Inglés | MEDLINE | ID: mdl-32731380

RESUMEN

Microalgae productive chains are gaining importance as sustainable alternatives to obtain natural pigments. This work presents a review on the most promising pigments and microalgal sources by gathering trends from a 10-year bibliometric survey, a patents search, and an industrial and market analysis built from available market reports, projects and companies' webpages. The performed analysis pointed out chlorophylls, phycocyanin, astaxanthin, and ß-carotene as the most relevant pigments, and Chlorella vulgaris, Spirulina platensis, Haematococcus pluvialis, and Dunaliella salina, respectively, as the most studied sources. Haematococcus is referred in the highest number of patents, corroborating a high technological interest in this microalga. The biorefinery concept, investment in projects and companies related to microalgae cultivation and/or pigment extraction is increasingly growing, particularly, for phycocyanin from Spirulina platensis. These pieces of evidence are a step forward to consolidate the microalgal pigments market, which is expected to grow in the coming years, increasing the prospects of replacing synthetic pigments by natural counterparts.


Asunto(s)
Industria Farmacéutica , Microalgas/química , Industria Farmacéutica/economía , Industria Farmacéutica/tendencias , Ficocianina/química , Ficocianina/aislamiento & purificación , Pigmentos Biológicos/química , Pigmentos Biológicos/economía , Pigmentos Biológicos/aislamiento & purificación
11.
Int J Biol Macromol ; 153: 256-263, 2020 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-32142842

RESUMEN

The phycocyanin was purified by Sephadex- G-100 and RP-HPLC and protein content was found to be 52.82% and the high purity fraction was collected and RP-HPLC analysis of fractionated phycocyanin, the α-subunit and ß-subunit were detected in 4.9 and 11.1(mAU). The frequency of peak 1456.26 cm-1 has showed the CH2 bending vibration and the protein amide II band was detected at 1539.20 cm-1 (CO stretching) and 2358.94 cm-1. In 1H NMR analysis, 14 chemical shifts (δ) were observed and signals confirmed namely alkyl halide, alkene, aldehyde proton and carboxylic acid. The in vivo anticancer effect was assessed by MTT assay against HepG-2 cell lines and in vivo antidiabetic effect was carried out through α-amylase and ß-glucosidase enzyme inhibition methods. The promising anticancer effect 68% was noticed at the concentration of 500 µg/ml and lower anticancer effect was noticed at the concentration of 100 µg/ml against Hep-G2 cell lines. The α-amylase and ß-glucosidase enzyme inhibition of phycocyanin showed dose dependent and maximum inhibition effect at 250 µg/ml. Phycocyanin anti-inflammatory effect such as inhibition of albumin denaturation, antiproteinase, hypotonicity-induced haemolysis and anti-lipoxygenase activities have been recorded maximum level at 500 µg/ml. Phycocyanin have complex structure and high molecular weight with more biomedical applications for drug development.


Asunto(s)
Ficocianina/química , Ficocianina/farmacología , Spirulina/química , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/farmacología , Antineoplásicos/química , Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Chlorocebus aethiops , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/aislamiento & purificación , Inhibidores Enzimáticos/farmacología , Células Hep G2 , Humanos , Hipoglucemiantes/química , Hipoglucemiantes/aislamiento & purificación , Hipoglucemiantes/farmacología , Ficocianina/aislamiento & purificación , Células Vero , alfa-Amilasas/antagonistas & inhibidores , beta-Glucosidasa/antagonistas & inhibidores
12.
Int J Biol Macromol ; 150: 38-51, 2020 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-32035961

RESUMEN

C-phycoyanins are abundant light-harvesting pigments which have an important role in the energy transfer cascade of photosystems in prokaryotic cyanobacteria and eukaryotic red algae. These proteins have important biotechnological applications, since they can be used in food, cosmetics, nutraceutical, pharmaceutical industries and in biomedical research. Here, C-phycocyanin from the extremophilic red alga Galdieria phlegrea (GpPC) has been purified and characterized from a biophysical point of view by SDS-PAGE, mass spectrometry, UV-Vis absorption spectroscopy, circular dichroism and intrinsic fluorescence. Stability against pH variations, addition of the oxidizing agent hydrogen peroxide and the effects of temperature have been also investigated, together with its in cell antioxidant potential and antitumor activity. GpPC is stable under different pHs and unfolds at a temperature higher than 80 °C within the pH range 5.0-7.0. Its fluorescence spectra present a maximum at 650 nm, when excited at 589 nm. The protein exerts interesting in cell antioxidant properties even after high temperature treatments, like the pasteurization process, and is cytotoxic for A431 and SVT2 cancer cells, whereas it is not toxic for non-malignant cells. Our results assist in the development of C-phycocyanin as a multitasking protein, to be used in the food industry, as antioxidant and anticancer agent.


Asunto(s)
Fenómenos Químicos , Ficocianina/química , Secuencia de Aminoácidos , Antineoplásicos/química , Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Cromatografía Liquida , Concentración de Iones de Hidrógeno , Ficocianina/aislamiento & purificación , Ficocianina/farmacología , Estabilidad Proteica , Especies Reactivas de Oxígeno/metabolismo , Rhodophyta/química , Espectrometría de Masas en Tándem , Temperatura
13.
Food Chem ; 316: 126374, 2020 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-32066073

RESUMEN

Arthrospira platensis, commonly known as Spirulina, gains increasing importance as alternative protein source for food production and biotechnological systems. A promising area is functional high-value algae extracts, rich in phycocyanin, a protein-pigment complex derived from A. platensis. This complex has proven functionality as the only natural blue colorant, fluorescent marker and therapeutic agent. The structure-function relationship is heat sensitive, making thermal processing in its production and its subsequent application a crucial aspect. In continuous high-temperature short-time treatments, it was shown how a purified phycocyanin (mixture of allophycocyanin and c-phycocyanin) disassembled and denatured between 50 and 70 °C. Three characteristic transition temperatures were allocated to specific quaternary aggregates. In contrast to sequential chemical denaturation, phycocyanin's chromophore and protein structure were simultaneously affected by thermal processing. Through a functionality assessment, the findings help optimize the efficiency of raw material usage by defining a processing window, enabling targeted process control resulting in desired product properties.


Asunto(s)
Ficocianina/química , Spirulina/química , Dicroismo Circular , Color , Ficocianina/aislamiento & purificación , Temperatura , Factores de Tiempo
14.
Microbiologyopen ; 9(3): e989, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-31970933

RESUMEN

Phycobiliproteins (PBPs) are colored fluorescent proteins present in cyanobacteria, red alga, and cryptophyta. These proteins have many potential uses in biotechnology going from food colorants to medical applications. Allophycocyanin, the simplest PBP, is a heterodimer of αß subunits that oligomerizes as a trimer (αß)3 . Each subunit contains a phycocyanobilin, bound to a cysteine residue, which is responsible for its spectroscopic properties. In this article, we are reporting the expression of recombinant allophycocyanin (rAPC) from the eukaryotic red algae Agarophyton chilensis in Escherichia coli, using prokaryotic accessory enzymes to obtain a fully functional rAPC. Three duet vectors were used to include coding sequences of α and ß subunits from A. chilensis and accessorial enzymes (heterodimeric lyase cpc S/U, heme oxygenase 1, phycocyanobilin oxidoreductase) from cyanobacteria Arthrospira maxima. rAPC was purified using several chromatographic steps. The characterization of the pure rAPC indicates very similar spectroscopic properties, λmaxAbs , λmaxEm , fluorescence lifetime, and chromophorylation degree, with native allophycocyanin (nAPC) from A. chilensis. This method, to produce high-quality recombinant allophycocyanin, can be used to express and characterize other macroalga phycobiliproteins, to be used for biotechnological or biomedical purposes.


Asunto(s)
Eucariontes/genética , Ficocianina/biosíntesis , Ficocianina/genética , Células Procariotas/enzimología , Proteínas Recombinantes , Electroforesis en Gel de Poliacrilamida , Expresión Génica , Vectores Genéticos/genética , Peso Molecular , Ficocianina/aislamiento & purificación , Análisis Espectral
15.
J Food Sci ; 85(3): 727-735, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31999367

RESUMEN

Phycocyanin, a natural blue colorant, is typically extracted from liquid biomass of Arthrospira platensis, a blue-green algae called spirulina. In this study, we developed a scalable process to extract phycocyanin from dried spirulina biomass. First, we established the optimal ionic strength and pH for the extraction buffer. The results showed that a minimum ionic strength (>5 g/L NaCl) must be maintained to minimize the co-extraction of the green chlorophyll. The optimal pH of the phosphate buffer (100 mM) for phycocyanin extraction is 7.5; however, the pH should be immediately adjusted to 6.0 to 6.5 after the extraction to keep phycocyanin stable. Second, we also investigated three processing techniques, that is, high-pressure processing (HPP), pulsed electric field (PEF), and ultrasonication, to break the cell walls of spirulina and facilitate the release of phycocyanins into extraction buffers. HPP and PEF do not lead to the release of phycocyanin into the extraction buffer. However, ultrasonication breaks down the spirulina into fine particles and releases most of the phycocyanin, along with other impurities, immediately after the treatment. Last, it has been revealed that most of the phycocyanin can be extracted from biomass within 3 hr by phosphate buffer only (pH 7.5, 100 mM) at room temperature. It is concluded that there is no need to treat the rehydrated biomass solution by HPP, PEF, or ultrasonication due to the minimal benefits they brought for the extraction. Based on these results, we proposed an extraction process for the plant production of phycocyanin starting from dried spirulina biomass. PRACTICAL APPLICATIONS: Limited information can be found on the extraction of phycocyanin from dried spirulina biomass, especially how to better preserve the natural blue color of phycocyanin during extraction. We have investigated the method and presented a different view from previous processes. Pulsed electric field, high-pressure processing, and ultrasonication were employed to accelerate the extraction of phycocyanin from dried biomass. However, it was found that, unlike the extraction from live wet biomass, these techniques do not help with the extraction from dried biomass. Based on investigations, we have proposed a process that can be easily scaled up for the commercial production of phycocyanin from dried spirulina biomass.


Asunto(s)
Fraccionamiento Químico/métodos , Colorantes de Alimentos/aislamiento & purificación , Manipulación de Alimentos/métodos , Ficocianina/aislamiento & purificación , Spirulina/química , Biomasa , Fraccionamiento Químico/instrumentación , Clorofila/análisis , Clorofila/aislamiento & purificación , Colorantes de Alimentos/análisis , Manipulación de Alimentos/instrumentación , Concentración de Iones de Hidrógeno , Ficocianina/análisis , Spirulina/crecimiento & desarrollo
16.
Methods Mol Biol ; 1980: 173-179, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-30484147

RESUMEN

Phycocyanin is a blue-colored pigment-protein complex that exhibits numerous biofunctions such as anti-inflammation, antioxidation, antitumor, neuroprotective effect, and immunological enhancement. Purified phycocyanin has pharmaceutical and nutraceutical applications. In addition, as a nontoxic and non-carcinogenic natural coloring agent, phycocyanin has many applications in the food and cosmetic industries. This chapter describes a protocol for extraction and analytical grade purification of phycocyanin from cyanobacteria. The purification steps include (1) extraction of phycocyanin from biomass, (2) ammonium sulfate precipitation of phycocyanin and dialysis, and (3) purification of phycocyanin by gel filtration and ion-exchange chromatography.


Asunto(s)
Cianobacterias/metabolismo , Ficocianina/biosíntesis , Ficocianina/aislamiento & purificación , Sulfato de Amonio , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Precipitación Fraccionada , Extracción Líquido-Líquido , Ficocianina/análisis
17.
Appl Microbiol Biotechnol ; 103(23-24): 9455-9464, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31696285

RESUMEN

The setup of an economic and sustainable method to increase the production and commercialization of products from microalgae, beyond niche markets, is a challenge. Here, a cascade approach has been designed to optimize the recovery of high valuable bioproducts starting from the wet biomass of Galdieria phlegrea. This unicellular thermo-acidophilic red alga can accumulate high-value compounds and can live under conditions considered hostile to most other species. Extractions were performed in two sequential steps: a conventional high-pressure procedure to recover phycocyanins and a solvent extraction to obtain fatty acids. Phycocyanins were purified to the highest purification grade reported so far and were active as antioxidants on a cell-based model. Fatty acids isolated from the residual biomass contained high amount of PUFAs, more than those recovered from the raw biomass. Thus, a simple, economic, and high effective procedure was set up to isolate phycocyanin at high purity levels and PUFAs.


Asunto(s)
Ácidos Grasos/aislamiento & purificación , Ficocianina/aislamiento & purificación , Rhodophyta/química , Biomasa , Biotecnología/métodos , Ácidos Grasos/metabolismo , Ficocianina/metabolismo
18.
Sci Rep ; 9(1): 9474, 2019 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-31263160

RESUMEN

C-phycocyanin (C-PC) pigment, as a natural blue dye, has particular applications in various fields. It is a water-soluble protein which has anticancer, antioxidant and anti-inflammatory properties. Here, we introduce an efficient procedure for the purification of C-PC pigment, followed by conducting a comprehensive investigation of its cytotoxic effects on human breast cancer (MCF-7) cells and the underlying mechanisms. A novel four-step purification procedure including the adsorption of impurities with chitosan, activated charcoal, ammonium sulfate precipitation, and ion exchange chromatography was employed, achieving a high purity form of C-PC with purity index (PI) of 5.26. SDS-PAGE analysis showed the purified C-PC with two discrete bands, subunit α (17 kD) and ß (20 kD), as confirmed its identity by Native-PAGE. A highly purified C-PC was employed to evaluate its anticancer activity and underlying molecular mechanisms of action. The inhibitory effects of highly purified C-PC on the proliferation of human breast cancer cells (MCF-7) have detected by MTT assay. The IC50 values for 24, 48, and 72 hours of exposure to C-PC were determined to be 5.92, 5.66, and 4.52 µg/µl, respectively. Flow cytometric analysis of cells treated with C-PC, by Annexin V/PI double staining, demonstrated to induce MCF-7 cells apoptosis. Also, the results obtained from propidium iodide (PI) staining showed that MCF-7 cells treated with 5.92 µg/µl C-PC for 24 h would arrest at the G2 phase and 5.66 and 4.52 µg/µl C-PC for 48 and 72 h could induce cell cycle arrest at both G2 and S phases. The oxidative damage and mitochondrial dysfunction were evaluated to determine the possible pathways involved in C-PC-induced apoptosis in MCF-7 cells. Our findings clearly indicated that the treatment of MCF-7 cells with C-PC (IC50 for 24 h) increased the production of reactive oxygen species (ROS). Consequently, an increase in the lipid peroxidation (LPO) level and a reduction in the ATP level, mitochondrial membrane potential (MMP), glutathione (GSH) and its oxidized form (GSSG), occurred over time. The reduced expression levels of anti-apoptotic proteins, Bcl2 and Stat3, plus cell cycle regulator protein, Cyclin D1, using Real-Time PCR confirm that the C-PC-induced death of MCF-7 human breast cancer cells occurred through the mitochondrial pathway of apoptosis. Collectively, the analyses presented here suggest that C-PC has the potential so that to develop it as a chemotherapeutic anticancer drug.


Asunto(s)
Antineoplásicos , Proteínas Bacterianas , Cianobacterias/química , Neoplasias/tratamiento farmacológico , Ficocianina , Antineoplásicos/química , Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Proteínas Bacterianas/química , Proteínas Bacterianas/aislamiento & purificación , Proteínas Bacterianas/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Células MCF-7 , Neoplasias/metabolismo , Neoplasias/patología , Ficocianina/química , Ficocianina/aislamiento & purificación , Ficocianina/farmacología
19.
Int J Biol Macromol ; 137: 647-656, 2019 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-31265852

RESUMEN

In our study, we focused on the optimization; antioxidant and hepatoprotective potentials of novel pigment-protein complex(C-PC) isolated from Phormidium versicolor against cadmium induced liver injury in rats. From analysis, the C-PC extraction parameters were optimized using the response surface methodology (RSM) for optimal recoveries of C-PC extraction. For analysis, the optimum operational conditions for maximizing phycocyanins concentration (67.45mg/g DM) were found to be water/solid 2, temperature 32.5°C and pH7.2.This pigment was identified using HPLC and FTIR analysis. In addition, the molecular masses of α and ß subunits were 17 and 19kDa. Scavenging activity of superoxide anion, hydroxyl, nitric oxide radicals and metal chelating in vitro results indicated that C-PC has an excellent capacity as antioxidant. In vivo study, C-PC significantly prevented cadmium-induced elevation of ALAT, ASAT and bilirubin levels in rats. The histopathological observations supported the results serum enzymes assays. The results of this study revealed that C-PC has significant hepatoprotective potential. C-PC (50mgkg-1 body weight) significantly enhanced the levels of antioxidant enzymes. It can be concluded that C-PC possesses prevention action against hepatotoxicity caused by cadmium.


Asunto(s)
Biotecnología/métodos , Citoprotección/efectos de los fármacos , Hígado/efectos de los fármacos , Microalgas/metabolismo , Ficocianina/biosíntesis , Ficocianina/farmacología , Animales , Antioxidantes/aislamiento & purificación , Antioxidantes/metabolismo , Antioxidantes/farmacología , Cadmio/toxicidad , Glicosilación/efectos de los fármacos , Concentración de Iones de Hidrógeno , Hígado/citología , Hígado/lesiones , Hígado/metabolismo , Ficocianina/aislamiento & purificación , Ratas , Temperatura
20.
Int J Biol Macromol ; 135: 62-68, 2019 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-31121226

RESUMEN

Phycocyanin (PC) is the principal pigment protein in the light-harvesting antenna of cyanobacteria. Here the biochemical characterization and the 1.51 Šcrystal structure of PC from cyanobacterium Nostoc sp. WR13 (Nst-PC) is reported. The P63 crystal lattice is composed of the minimal biological entities of Nst-PC, the (αß)3 trimeric rings. The structure has been refined to R factor 11.5% (Rfree 15.4%) using anisotropic atomic B factors. A phylogenetic study shows that the α and ß chains of Nst-PC are significantly clustered in a distinct clade with Acaryochloris marina. The structure was examined to look for any significant differences between Nst-PC and PC from non-desert species. Only minor differences were found in the chromophore microenvironments. The tentative energy transfer pathways in Nst-PC were modeled based on simple structural considerations.


Asunto(s)
Modelos Moleculares , Estructura Molecular , Nostoc/enzimología , Ficocianina/química , Cristalografía por Rayos X , Transferencia de Energía , Conformación Molecular , Ficobiliproteínas/química , Ficobilisomas/química , Ficocianina/aislamiento & purificación , Análisis Espectral
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