Development of a process for large-scale purification of C-phycocyanin from Synechocystis aquatilis using expanded bed adsorption chromatography

Ramos, Amparo; Acién, F.G.; Sevilla, J. M. Fernández; González, Cynthia V.; Bermejo, Ruperto

doi:10.1016/j.jchromb.2011.01.013

Cited by 49 publications

(22 citation statements)

References 35 publications

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“…In comparison with previous papers that describe similar methods [13][14][15][16][17][18][19][20][21] for phycobiliproteins purification using expanded bed chromatography, we find important differences. So, Niu et al [19] described a method based on EBAC, using a 25 mm ID column.…”

Section: Recovery Evaluationcontrasting

confidence: 61%

“…The PCs rich solution from EBAC step was used to obtain pure C-PC by anion-exchange chromatography [21]. The content and purity of C-PC purified by anion-exchange chromatography was determined by UV-Vis absorption spectroscopy, steady-state fluorescence and electrophoresis.…”

Section: Purification and Identification Of C-phycocyaninmentioning

confidence: 99%

“…This technique can be used in general primary protein purification [12] and more specifically in phycobiliproteins purification. In this sense, there are different studies about phycoerythrins [13][14][15][16][17] and phycocyanins [18][19][20][21] purification using EBAC at small, medium and large scale obtaining diverse protein quantities, but always in the order of milligrams.…”

Section: Introductionmentioning

confidence: 99%

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Pilot Scale Recovery of Phycocyanin from Spirulina platensis Using Expanded Bed Adsorption Chromatography

Bermejo

Ramos

2012

Chromatographia

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The use of expanded bed adsorption chromatography (EBAC) for pilot scale recovery of C-phycocyanin (C-PC) from the cyanobacteria Spirulina platensis is described. The description includes the whole procedure, from small-scale method optimization to pilot scale. Initially, the phycobiliproteins are extracted by osmotic shock and separated by applying the centrifuged cell suspension to an EBAC column, using Streamline-DEAE as adsorbent. Next, the PCs rich solution is eluted to finally be purified by packed bed chromatography using DEAE-cellulose. The EBAC operation was optimized on a small scale using a 15 mm internal diameter (ID) column. The optimal conditions were: a sample load of 0.91 mg C-PC mL -1 adsorbent, H/H 0 = 2 and a sample viscosity of 1.084 mP. The EBAC process was then scaled up by increasing the column ID (15, 25, 40, 60 and 90 mm) and, finally, the process was developed at pilot scale using a 150 mm ID column obtaining a scale-up factor of 100. The yield of the EBAC step is in the range of 75-79% whatever the column diameter. To obtain pure C-PC, conventional chromatography with DEAE-cellulose was utilized. The overall yield of the purification process after all steps is 59%. The purification steps are monitored using SDS-PAGE and the purity of recovered C-PC is confirmed by absorption and emission spectroscopy. Results show that EBAC method is a scalable technology that allows large quantities of C-PC to be obtained without product loss, maintaining a high protein recovery while reducing both processing cost and times.

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Section: Recovery Evaluationcontrasting

confidence: 61%

Section: Purification and Identification Of C-phycocyaninmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Pilot Scale Recovery of Phycocyanin from Spirulina platensis Using Expanded Bed Adsorption Chromatography

Bermejo

Ramos

2012

Chromatographia

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“…Silveira et al (2008) , respectively. The values for Q m of the same support and target productStreamline DEAE and C-phycocyanin, respectivelyfor different cyanobacteria are variable, and values for Q m of 0.8 mg.mL −1 (Bermejo et al, 2006), 1.6 mg.mL -1 (Bermejo and Ramos, 2012), 1.74 mg.mL −1 (Ramos et al, 2010) and 5.2 mg.mL −1 (Ramos et al, 2011) can be found in the literature. Thus the Streamline DEAE resin showed maximum adsorption 7 times greater than the maximum value mentioned above, reflecting the importance of studying the adsorption conditions better, for instance the influence of pH and temperature.…”

Section: Adsorption Isothermmentioning

confidence: 99%

“…Silveira et al (2008) studied the effects of pH and temperature on the partition coefficient, and determined the adsorption isotherm under the best conditions evaluated for the adsorption of C-phycocyanin onto the ion exchange resin Q-Sepharose FF, a widely used and highly successful resin applied in fixed beds. Adsorption in an expanded bed resin of Streamline DEAE was studied by Bermejo et al (2006), Ramos et al (2010), Ramos et al (2011) and Bermejo and Ramos (2012), obtaining the adsorption isotherms for crude C-phycocyanin and/or a purified extract to obtain the parameter Q m . In these studies the kinetic behavior was not evaluated and the pH and temperature were not optimized for better adsorption.…”

Section: Introductionmentioning

confidence: 99%

Kinetics and adsorption isotherm of C-phycocyanin from Spirulina platensis on ion-exchange resins

Sala

Figueira

Cerveira

et al. 2014

Braz. J. Chem. Eng.

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-C-phycocyanin is a natural blue dye extracted from Spirulina platensis, which has many applications in the food and pharmaceutical industries. In this paper the effect of pH and temperature on the adsorption of C-phycocyanin onto two different ion exchange resins (Streamline DEAE and Streamline Q XL) for expanded bed adsorption chromatography was investigated. Moreover, the kinetics and adsorption isotherm were evaluated. The equilibrium for the Q XL matrix was reached after 60 min, while for DEAE it was only reached after 140 min. C-phycocyanin showed the highest partition coefficient at pH 7.5 for both resins at 25 °C. The C-phycocyanin adsorption isotherm was very well represented by the Langmuir, Freundlich and Langmuir-Freundlich models, where the estimated values for Q m and K d obtained by the Langmuir isotherm were, respectively, 33.92 mg.mL -1 and 0.123 mg.mL -1 for DEAE, and 28.12 mg.mL -1 and 0.082 mg.mL -1 for the Q XL matrix. A negative cooperativity was observed for C-phycocyanin binding when the Q XL matrix was used, while the cooperativity was purely independent using the DEAE matrix.

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Phycocyanins

Bermejo

2013

Cyanobacteria

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Development of a process for large-scale purification of C-phycocyanin from Synechocystis aquatilis using expanded bed adsorption chromatography

Cited by 49 publications

References 35 publications

Pilot Scale Recovery of Phycocyanin from Spirulina platensis Using Expanded Bed Adsorption Chromatography

Pilot Scale Recovery of Phycocyanin from Spirulina platensis Using Expanded Bed Adsorption Chromatography

Kinetics and adsorption isotherm of C-phycocyanin from Spirulina platensis on ion-exchange resins

Phycocyanins

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