No description available.

Phycocyanin (PC), a pigment-protein conjugate from Arthrospira platensis, is increasingly used in foods as a natural alternative to artificial blue dyes. Although PC has been classified as a color additive exempt from certification by the Food and Drug Administration, its limited stability has hindered its widespread application in food products. The objectives of this study were: a) to evaluate the photophysical properties of PC and their sensitivity to temperature, viscosity, and water activity, b) to monitor PC’s thermal degradation based on changes in the optical properties of its intrinsic fluorophores, namely its chromophores and aromatic amino acids, and c) to extract PC’s thermal degradation kinetics parameters from non-isothermal degradation profiles and validate their predictive ability. PC’s photophysical properties were monitored in solutions with viscosities from 1 to8000 mPa s and water activities, aw, from about 0 to 1. PC’s emission intensity showed high sensitivity to aw above 0.8 and mild sensitivity to the viscosity of its local environment. The effect of temperature on PC’s photophysical properties was tested in aqueous PC solutions (0.5 mM, pH: 6.1) subjected to non-isothermal temperature profiles with target temperatures from 42.5 to 80°C. The stability of PC was monitored in terms of its photophysical properties, i.e., fluorescence emission intensity, energy, and anisotropy (r) of its chromophore at set time intervals. Additionally, the photophysical properties of PC’s aromatic amino acids (AAs) tyrosine and tryptophan (lexc: 280 and 295 nm) were recorded. The thermal degradation kinetics of PC was assumed to follow a Weibullian model, and the temperature dependence of the degradation rate parameter, b(T), a logarithmic exponential model. Changes of PC fluorescence intensity under dynamic conditions were used to extract the degradation kinetics parameters using the endpoints method. Deviations between the estimated and experimental values were less than 10% for all temperature profiles. During thermal treatments, hypsochromic shifts of AAs’ emission spectra (from 340 to 315 nm) and significant increases in fluorescence anisotropy revealed that color losses were not solely associated with an alteration of the chromophore but with conformational changes and possible aggregation of the protein subunits. An increase in viscosity of the surrounding media provided a protected effect on discoloration during heating. Adequate modeling approaches and molecular spectroscopic techniques can help to develop effective strategies to enhance thermal stability, expand its use as a color and functional ingredient and operationalize it as an endogenous sensor of food quality.

The predicted protein product of open reading frame slr0328 from Synechocystis sp. PCC 6803, SynPTP, possesses significant amino acid sequence similarity with known low molecular weight protein tyrosine phosphatases (PTPs). To determine the gross functional properties of this hypothetical protein, open reading frame slr0328 was cloned, and its predicted protein product was expressed in E. coli. The recombinant protein, SynPTP, was purified by metal ion column chromatography. The catalytic activity of SynPTP was examined toward several exogenous protein substrates that had been phosphorylated on either tyrosine residues or serine residues. SynPTP exhibited phosphatase activity toward tyrosine phosphorylated protein substrates (Vmax toward phosphotyrosyl 32P-casein was 1.5 nmol/min/mg). However, no phosphatase activity was detected toward serine phosphorylated protein substrates. SynPTP displayed phosphohydrolase activity toward several organophosphoesters including para-nitrophenyl phosphate (p-NPP), beta-napthyl phosphate and phosphotyrosine but not toward alpha-napthyl phosphate, phosphoserine, or phosphothreonine. Kinetic analysis indicated that the Km (0.6 mM) and Vmax (3.2 mmole/min/mg) values for SynPTP toward pNPP are similar to those of other known bacterial low molecular weight PTPs. The protein phosphatase activity of SynPTP was inhibited by sodium orthovanadate, a known inhibitor for tyrosine phosphatases, but not by okadaic acid, an inhibitor for many serine/threonine phosphatases. Mutagenic alteration of the predicted catalytic cysteine, Cys7, to serine abolished enzyme activity. Several phosphotyrosine containing proteins were detected from the whole cell extracts of Synechocystis sp. PCC 6803 through immunoreactions using anti-phosphotyrosine antibody. SynPTP was observed to dephosphorylate three of these proteins in vitro. Two of these proteins were identified by peptide-mass fingerprinting analysis, as PsaD (photosystem I subunit II) and CpcD (phycocyanin rod linker protein). In addition, several phosphotyrosine proteins were detected from the soluble and membrane fractions of Synechocystis sp. PCC 6803 cell extracts by in vitro substrate trapping as potential endogenous substrates of SynPTP. Two of these proteins were identified as the alpha and beta subunits of phycocyanin. We therefore speculate that SynPTP might be involved in the regulation of photosynthesis in Synechocystis sp. PCC 6803. / Ph. D.

The Western Mississippi Sound (WMS) is home to the largest natural oyster reef in the Gulf of Mexico and contributes substantially to Mississippi's economy. In 2019, the WMS experienced an unprecedented cyanobacterial bloom that killed fish and birds and led to shut down of beaches and oyster fishery. This thesis aims to quantify cyanobacteria from uncrewed aircraft systems (UAS) imagery and investigate the relative influence of river discharge into the WMS on cyanobacterial concentrations. Several field campaigns were undertaken to collect field data and UAS imagery from WMS. A remote sensing algorithm was developed to quantify the unique cyanobacterial pigment phycocyanin and generate temporal maps for cyanobacteria using UAS imagery. Correlations between the cyanobacteria maps and discharge of major freshwater sources to WMS revealed that Bonnet Carré Spillway had the largest contribution followed by discharge of Jourdan, Wolf, and Pearl rivers to the cyanobacterial concentrations over the oyster reef.

Cyanobacteria are photosynthetic organisms that efficiently capture light by utilizing the light-harvesting complexes called phycobilisomes. In many cyanobacteria, phycobilisomes are composed of an allophycocyanin core with phycocyanin and phycoerythrin (PE) rods radiating from the core. These phycobiliproteins have multiple bilin chromophores, such as phycoerythrobilin (PEB), covalently attached to specific cysteine (Cys) residues for efficient photosynthetic light capture. Chromophore ligation on phycobiliprotein subunits occurs through bilin lyase catalyzed reactions. This study mainly focuses on characterizing the roles of enzymes that are involved in the biosynthetic pathway of the phycobiliproteins within two cyanobacteria Thermosynechococcus elongatus and Fremyella diplosiphon. A combination of molecular and biochemical techniques were used to better understand the roles of these proteins in the post-translational modification and/or stability of phycobiliproteins. Using a heterologous plasmid coexpression system in E. coli, recombinant CpcS-III from T. elongatus was shown to ligate three different bilins to both subunits of allophycocyanin and to the beta subunit of phycocyanin, thus, acting as a bilin lyase. The crystal structure of CpcS-III was also solved, the first bilin lyase structure. Next, the roles of three proteins from F. diplosiphon CpeY, CpeZ, and CpeF were analyzed using a combination of gene knock-out mutants and recombinant protein expression techniques. In the absence of cpeY, chromophorylation to the alpha subunit of PE at Cys-82 was reduced, coinciding with the recombinant data that CpeY is the lyase that attaches PEB to this site. Removing cpeZ from the genome resulted in the destabilization and reduced accumulation of PE, especially the beta subunit CpeB. Recombinant CpeZ was shown to act like a chaperone-like protein and increased the solubility and fluorescence of both recombinant and native CpeB by increasing the stability of the phycobiliprotein and/or by increasing the activities of other lyases. The deletion of cpeF resulted in a reduced-PE phenotype with the doubly attached PEB missing from CpeB at Cys-48/Cys-59. Recombinant CpeF was shown to ligate PEB to CpeB-Cys-48/Cys-59 in the presence of recombinant CpeS (lyase attaches PEB to CpeB-Cys-80) and CpeZ. CpeF also showed a chaperone-like function by stabilizing CpeB, but its main role appears to be as a bilin lyase.

Fundação de Amparo a Pesquisa do Estado de São Paulo / Cyanobacteria are prokaryote microorganisms that gather interesting features for the biotechnological use in biorefinery systems. Phormidium sp. is a cyanobacterium that has metabolic versatility, grown in both photosynthetic and heterotrophic conditions, from industrial residues, generating a chemically attractive biomass. On face of it, the work aimed to evaluate the production of high added value chemicals by Phormidium sp. from agro-industrial residues. It focuses on (i) identification and quantification of carotenoids from heterotrophically grown biomass in agro-industrial wastewater (ii) identification and quantification of carotenoids from photoautotrophically grown biomass and evaluation of the antioxidant capacity of the extract (iii) quantification of phycobiliproteins from photoautotrophically grown biomass and evaluation of the antioxidant capacity of the extract (iv) quantification of chlorophyll from photoautotrophically grown biomass and the evaluation of the antioxidant capacity of the extract. The results showed that the microalgal biomass from heterotrophic growth had 183.03 ± 0.9 g.g-1 of total carotenoids while in the biomass from photosynthetic growth this value was 714.3 ± 0.9 g.g-1. The major carotenoids in both extracts were all-trans--carotene, all-trans-lutein and all-trans-zeaxanthin. The carotenoid extract was able to scavenge the peroxyl radical 28 times more than the -tocopherol standard. The content of phycocyanin found was 20.05 ± 1.1 g.100g-1 and after fractionation achieved a purity ratio of 4.8 as well as antioxidant capacity of 237.4 ± 0.7 mol trolox equivalents.g-1. In terms of chlorophyll, the biomass showed the content of 3400 g.g-1 and the capacity to scavenge the peroxyl radical 84.9 in relation to -tocopherol standard. The results show that Phormidium sp. microalgal biomass has potential for biotechnological exploitation as a renewable and low cost source of bioactive compounds of high added value. / Cianobactérias são seres de estrutura procarionte que reúnem interessantes características para a utilização biotecnológica em sistemas de biorrefinaria. A Phormidium sp. é uma cianobactéria que possui uma versatilidade metabólica que a permite crescer em condições fotossintéticas e heterotróficas, partir de resíduos industriais, gerando uma biomassa quimicamente atrativa. Em face disto, o trabalho teve por objetivos avaliar a produção de químicos de alto valor por Phormidium sp. a partir de resíduos agroindustriais. O foco foi direcionado a (i) identificação e quantificação de carotenoides da biomassa obtida heterotroficamente a partir de efluente agroindustrial (ii) a identificação e quantificação de carotenoides da biomassa cultivada fotoautotroficamente e a avaliação da capacidade antioxidante do extrato (iii) quantificação das ficobiliproteínas da biomassa cultivada fotoautotroficamente a avaliação da capacidade antioxidante do extrato e (iv) quantificação das clorofilas da biomassa cultivada fotoautotroficamente a avaliação da capacidade antioxidante do extrato. Os resultados indicaram que a biomassa microalgal obtida do cultivo heterotrófico apresentou um teor de carotenoides de 183,03 ± 0,9 g.g-1 enquanto que na cultivada em condições fotossintéticas o teor foi de 714,3 ± 0,9 g.g-1. Os carotenoides majoritários em ambos os extratos foram all-trans--caroteno, all-trans-luteína e all-trans-zeaxantina. O extrato de carotenoides foi capaz de desativar o radical peroxila 28 vezes mais eficientemente que o padrão de -tocoferol. O conteúdo de ficocianina encontrado foi 20,05 ± 1.1 g.100g-1 biomassa e após o fracionamento apresentou uma pureza de 4,8 e atividade antioxidante de 237,4 ± 0.7 mol trolox.g-1 biomassa. Em termos de clorofila, foram encontradas concentrações de 3400 g.g-1 e capacidade de desativar o radical peroxila relativa ao -tocoferol de 84,9. Os resultados encontrados demonstram que a biomassa microalgal de Phormidium sp. possui potencial para exploração biotecnológica como uma fonte renovável e de baixo custo de compostos bioativos de alto valor agregado.

Indiana University-Purdue University Indianapolis (IUPUI) / Nuisance blue-green algal blooms contribute to aesthetic degradation of water resources and produce toxins that can have serious adverse human health effects. Current field-based methods for detecting blooms are costly and time consuming, delaying management decisions. Remote sensing techniques which utilize the optical properties of blue-green algal pigments (chlorophyll a and phycocyanin) can provide rapid detection of blue-green algal distribution. Coupled with physical and chemical data from lakes, remote sensing can provide an efficient method for tracking cyanobacteria bloom occurrence and toxin production potential to inform long-term management strategies. In-situ field reflectance spectra were collected at 54 sampling sites on two turbid, productive Indianapolis reservoirs using ASD Fieldspec (UV/VNIR) spectroradiometers. Groundtruth samples were analyzed for in-vitro pigment concentrations and other physical and chemical water quality parameters. Empirical algorithms by Gitelson et al. (1986, 1994), Mittenzwey et al. (1991), Dekker (1993), and Schalles et al. (1998), were applied using a combined dataset divided into a calibration and validation set. Modified semi-empirical algorithms by Simis et al. (2005) were applied to all field spectra to predict phycocyanin concentrations. Algorithm accuracy was tested through a least-squares regression and residual analysis. Results show that for prediction of chlorophyll a concentrations within the range of 18 to 170 ppb, empirical algorithms yielded coefficients of determination as high as 0.71, RMSE 17.59 ppb, for an aggregated dataset (n=54, p<0.0001). The Schalles et al. (2000) empirical algorithm for estimation of phycocyanin concentrations within the range of 2 to 160 ppb resulted in an r2 value of 0.70, RMSE 23.97 ppb (n=48, p<0.0001). The Simis et al. (2005) semi-empirical algorithm for estimation of chlorophyll a and phycocyanin concentrations yielded coefficients of determination of 0.69, RMSE 20.51 ppb (n=54, p<0.0001) and 0.85, RMSE 24.61 pbb (n=49, p<0.0001), respectively. Results suggest the Simis et al. (2005) algorithm is robust, where error is highest in water with phycocyanin concentrations of less than 10 ppb and in water where chlorophyll a dominates (Chl:PC>2). A strong correlation between measured phycocyanin concentrations and blue-green algal biovolume measurements was also observed (r2=0.95, p<0.0001).

[ES] La inminente necesidad de un desarrollo sostenible ha influido en el crecimiento de la industria, abriendo el camino para la transición progresiva hacia una bioeconomía sostenible. La evolución de la industria textil, caracterizada por altos niveles de contaminación, exige la continua exploración de alternativas a los recursos fósiles y procesos contaminantes. En este sentido, el presente estudio propone la exploración de una alternativa sostenible de colorantes naturales procedentes de biomasa de algas, para su aplicación en procesos de coloración, seleccionados por su reducido impacto ambiental, tanto en el cultivo de las algas, como en la extracción de colorantes, y su posible aplicación textil. Este estudio tiene como objetivo explorar la viabilidad del uso de colorantes procedentes de algas para su aplicación en procesos de coloración textil, como la tintura por agotamiento y estampación pigmentaria. La selección de micro y macroalgas ha sido realizada según criterios que facilitan un cultivo optimizado y simple, ofreciendo diferentes colores básicos para la industria textil, como son el azul (C-ficocianina), el rojo (R-ficoeritrina), el amarillo (ß-caroteno), y el verde (Clorofila a). Los extractos líquidos concentrados en colorantes se han empleado en los procesos seleccionados de acabado textil, para conferir color a fibras naturales, como el algodón y la lana. La viabilidad de las fuentes alternativas de colorantes sostenibles mencionadas anteriormente ha sido estudiada mediante el análisis de la influencia de ciertos auxiliares de coloración, como son los mordientes. Una serie de mordientes convencionales (metálicos, y recién descubiertos) y biomordientes han sido seleccionados para investigar su capacidad de mejora de las propiedades y eficacia de coloración de los colorantes sostenibles. Asimismo, se han estudiado aproximaciones teóricas relacionados con la posible unión fibras-mordientes-colorantes. La implicación del uso de mordientes en los procesos de tintura por agotamiento y estampación pigmentaria se ha caracterizado a través de la medición de las coordenadas cromáticas del espacio de color; el espectro de reflectancia junto con la fuerza del color. El análisis se ha completado con el cálculo del coeficiente de absorción, en base al colorante remanente en los efluentes del agua residual de tintura (para los procesos de tintura). No obstante, la influencia de los mordientes sobre la calidad de los procesos de coloración ha sido evaluada mediante ensayos de medición de solideces al lavado y a la luz realizados según normativas europeas en vigor. El proceso de tintura por agotamiento se ha optimizado para determinar los valores óptimos para la temperatura, relación de baño de tintura, pH, tiempo de proceso, teniendo en cuenta la sensibilidad de los colorantes frente a los factores degradantes y de proceso. Siguiendo el carácter sostenible del proceso, se ha realizado una evaluación preliminar de los efluentes de aguas residuales de tinturas, a través de mediciones de indicadores como DBO5, DQO, contenido en metales, y el comportamiento frente a un tratamiento biológico de aguas residuales. Igualmente, se investigó el posible valor añadido de los colorantes estudiados, midiendo la capacidad de protección solar y antimicrobiana. El proceso de estampación pigmentaria abordó la investigación de la viabilidad de los colorantes mediante el empleo de la pasta de estampación sintética convencional, completada con la evaluación de la alternativa representada por la pasta de estampación natural, considerando, al mismo tiempo, la disponibilidad comercial de los componentes. Los resultados obtenidos confieren una validación preliminar de la idoneidad de la aplicación de materia colorante procedente de algas en procesos de tintura por agotamiento y la estampación pigmentaria sobre sustratos textiles de algodón y lana, a escala de laboratorio, con elementos clave para las perspectivas de funcionalización textil, abriendo así el camino para futuras investigaciones que hará posible el escalado de los procesos y el uso industrial de materiales alternativos sostenibles en la industria textil. / [CA] La creixent necessitat per a un desenvolupament sostenible ha tingut un gran impacte en el creixement de la indústria, obrint el camí per a la transició progressiva cap a una bioeconomia sostenible. L'evolució de la indústria tèxtil, caracteritzada per els alts nivells de contaminació fomenta la contínua exploració d'alternatives als recursos fòssils i processos contaminants. En aquest sentit, el present estudi proposa l'exploració d'una alternativa sostenible de colorants naturals procedents de biomassa d'algues, per a la seua aplicació en processos de coloració, seleccionats a causa de l'impacte ambiental reduït, generat pel cultiu de les algues, l'extracció de colorants, i la seua possible aplicació tèxtil. Aquest estudi té com a objectiu explorar la viabilitat de l'ús de colorants procedents d'algues per a la seua aplicació en processos de coloració tèxtil, com la tintura per esgotament i estampació pigmentària. La selecció de micro i macroalgues ha sigut realitzada segons criteris que faciliten un cultiu optimitzat i senzill, que proveeix colors bàsics per a la indústria tèxtil, com són el blau (C- ficocianina), el roig (R-ficoeritrina), el groc (ß-caroteno, i el verd (Clorofil·la a) . Els extractes líquids concentrats en colorants s'han emprat en els processos seleccionats d'acabat tèxtil, per a conferir color a fibres naturals, com el cotó i la llana. La viabilitat de les fonts alternatives de colorants sostenibles mencionades anteriorment ha sigut abordada per mitjà de l'anàlisi de la influència de certs auxiliars de coloració, com són els mordents. Una sèrie de mordents convencionals, metàl·lics, acabats de descobrir i bio mordents ha sigut seleccionada per a investigar la seua capacitat de millora de les propietats i eficàcia de coloració dels colorants sostenibles. A més a més, es va estudiar aproximacions teòriques relacionades amb la unió plausible entre fibres-mordents-colorants. La implicació de l'ús de mordents en els processos de tintura per esgotament i estampació pigmentària s'ha analitzat mitjançantassajos de caracterització, contemplant la caracterització objectiva del color per mitjà del mesurament de les coordenades cromàtiques de l'espai de color; l'espectre de reflectància junt amb la força del color; anàlisi completada pel càlcul del coeficient d'absorció, a base del colorant romanent en els efluents de l'aigua residual de tintura (per als processos de tintura). No obstant això, la influència dels mordents sobre la qualitat dels processos de coloració ha sigut avaluada per mitjà d'assajos de mesurament de solideses al llavat i a la llum realitzats segons normatives europees en vigor. El procés de tintura per esgotament ha sigut optimitzat per a determinar els valors òptims per a la temperatura, relació de bany de tintura, pH, temps de procés, valorant el caràcter sensible dels colorants front als agents degradants i de procés. Recolzant el caràcter sostenible del procés, s'ha realitzat una evaluación preliminar dels afluents d'aigües residuals de tintures, mitjançant mesuraments d'indicadors, com DBO5, DQO, contingut en metalls, i el comportament enfront d'un tractament biològic d'aigües residuals. Igualment, es va investigar el possible valor afegit dels colorants procedents d'algues, mesurant la capacitat de protecció solar i antimicrobiana. El procés d'estampació pigmentària va abordar la investigació de la viabilitat dels colorants per mitjà de l'ocupació de la pasta d'estampació sintètica convencional, completada amb l'avaluació de l'alternativa representada per la pasta d'estampació natural, considerant, al mateix temps, la disponibilitat comercial dels ingredients. Els resultats obtinguts confereixen una validació preliminar de la idoneïtat de l'aplicació de matèria colorant procedent d'algues en processos de tintura per esgotament i l'estampació pigmentària sobre substrats tèxtils de cotó i llana, a escala de laboratori, amb elements clau per a les perspectives de funcionalització tèxtil, obrint així el camí per a futures investigacions que farà possible l'escalat dels processos i l'ús industrial de materials alternatius sostenibles a la indústria tèxtil. / [EN] The crucial need for sustainable development has an immense impact on industrial evolution paving the path for a progressive transition towards a sustainable bioeconomy. Considering the pollutant evolution of the textile industry, approaches to exploring environmentally friendly alternatives to fossil-based resources and polluting processes have become the most current practice nowadays. In this sense, this study proposes the exploration of a sustainable alternative colorant material for coloration applications, from algae-based feedstock, considering the reduced environmental pressure embroiled by algae cultivation, colorant extraction, and its possible further textile application. This study explores the viability of the use of algae-based colorants for their application in exhaustion dyeing and pigment printing coloration techniques for the textile industry. Micro and macroalgae strains selection was performed according to criteria ensuring facile and optimized cultivation and the provision of basic colors for the textile industry as blue (C-phycocyanin), red (R-phycoerythrin), yellow (ß-carotene), and green (Chlorophyll-a). Liquid colorant-rich extracts were employed in the selected textile finishing processes on natural fibers, like cotton and wool. To approach the viability of these alternative sustainable colorants and enhance coloration properties and efficiencies, a series of auxiliaries were assessed, as conventional metallic and newly discovered mordants, and biomordants. Theoretical assumptions related to the plausible bonding among fibers-mordants-colorants were approximated. The auxiliaries implication in exhaustion dyeing and pigment printing processes were analyzed through a series of characterization tests, envisaging objective color characterization determined by the measurement of CIELab color coordinates; reflectance spectrum, and color strength approaching the depth of the coloration; completed by the calculation of the absorption coefficient, based on the remaining colorant matter in the wastewater effluents (for the dyeing process). Nevertheless, the mordants' influence on the quality of the coloration process was assessed through laundering and lightfastness measured according to European standards for textiles characterization. The dyeing exhaustion process was also subjected to parameters optimization in terms of the definition of optimum temperature, dyeing liquor ratio, pH, and time, considering the colorants' sensibilities for degrading and processing agents. A preliminary nearing towards the dyeing wastewater effluent quality assessment supporting the sustainability character of the process was performed through the measurement of basic water characterization indicators as BOD5, COD, metal content, and behavior against biological wastewater treatment. Furthermore, the possible added value of the algae-based colorants was investigated by measuring the antimicrobial and solar protection capacity, enlarging the application prospects of these innovative raw materials for textile applications. The pigment printing process approached the research of the feasibility of the colorants by employing the conventional synthetic printing paste, completed with the assessment of alternative natural printing paste, considering, at the same time the commercial availability of ingredients. The obtained results confer a preliminary validation of the suitability of the application of algae-based colorant matter in exhaustion dyeing and pigment printing of cotton and wool, at laboratory scale, with key elements for textile functionalization perspectives, thus paving the path for further investigation which will make possible the upscaling of the processes and industrial use of alternative sustainable materials in the textile industry. / Moldovan, S. (2021). Dyeing and Printing Natural Fibers with Algae-Based Colorants [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/180349

C-ficocianina (C-PC) é um pigmento solúvel em água e está presente em Arthrospira platensis. Este pigmento é constituído por duas subunidades, &#945 e &#946, com massas moleculares de 16 e 17 kDa, respectivamente. Recentemente foi demonstrado que a subunidade &#946 da C-PC de Anabaena tem atividade antitumoral. Os principais objetivos deste estudo foram clonar e expressar a subunidade &#946 de C-PC de A. platensis em amostras de Escherichia coli e verificar a capacidade da proteína recombinante e da proteína C-PC em induzir ou não apoptose em células tumorais (HEp-2) e não tumorais (fibroblastos). As células foram tratadas com 100 µg das proteínas recombinante e C-PC por 6 h. Após tratamento, as células foram coradas com azul de toluidina (Metódo: Concentração Crítica de Eletrólitos - CCE). As células em apoptose foram detectadas pela coloração e por alterações morfológicas. Os resultados obtidos demonstram que ambas as proteínas, &#946-PC recombinante e C-PC, são capazes de induzir apotose em células HEp-2 e não induzi-la em células fibroblásticas. / C-phycocyanin (C-PC) is a water soluble pigment and is present in Arthrospira platensis. It is consisted of two subunits, &#945 and &#946, with molecular masses of 16 and 17 kDa, respectively. Recently it was shown that the &#946 subunit of C-PC Anabaena has antitumor activity. The main objectives of this study were cloning and expression the &#946 subunit of C-PC (&#946-PC) of A. platensis in Escherichia coli and check the ability of the C-PC and recombinant proteins in apoptotic induction in cancer (HEp-2) and non-cancer (fibroblasts) cells. The fibroblasts and HEp-2 cells were treated with 100 µg of recombinant and C-PC, proteins respectively for 6 h. After treatment, the cells were stained with toluidine blue (Critical Electrolyte Concentration Method - CEC). Apoptosis cells were detected by staining and morphological changes. The results show that the recombinant &#946-PC and C-PC proteins are able to induce apotosis in HEp-2 cells and dont induce it in fibroblasts cells.

Thesis (M.S.)--Indiana University, 2007. / Department of Earth Sciences, Indiana University-Purdue University Indianapolis (IUPUI). Advisor(s): Li Lin, Lenore P. Tedesco, Jeffrey S. Wilson. Includes vitae. Includes bibliographical references (leaves 145-149).