Caroline Costa Moraes

C-phycocyanin extraction from Spirulina platensis wet biomass

C-Phycocyanin is a natural blue dye used in food and pharmaceutical industry. In the present study, a simple and efficient method to extract C-phycocyanin from Spirulina platensis wet biomass is reported. The extractions were carried out using six different methods, including chemical (organic and inorganic acid treatment), physical (freezing and thawing, sonication, homogenization) and enzymatic (lysozyme treatment) methods. The extraction using ultrasonic bath in the presence of glass pearls in the biomass proved to be the most efficient method, 56% higher than using freezing and thawing (the method most frequently used), and presented a extraction yield of 43.75 mg.g-1 and a C-phycocyanin concentration of 0.21 mg.mL-1.

Strategy for a protein purification design using C-phycocyanin extract.

A variety of techniques have been developed for the separation and recovery of proteins. The cost of purifying the product is frequently determined by the desired quality of the final product, which is evaluated by measuring the purity. In this work the design of a protein purification process for C-phycocyanin, a phycobiliprotein that can be used in the food and medical industries, was established. The study evaluated the use of ammonium sulfate precipitation, ion exchange chromatography and gel filtration to purify C-phycocyanin in a variety of sequences. The final design included the C-phycocyanin extraction step, precipitation with ammonium sulfate and ion exchange chromatography. When the elution step was studied, the kind of elution and pH were considered in order to obtain a product with a final purity of 4.0 with a purification factor of 6.35, so that, at the end of the strategy, C-phycocyanin of analytical grade would be obtained.

Mathematical modeling and simulation of inulinase adsorption in expanded bed column

A mathematical model for an expanded bed column was developed to predict breakthrough curves for inulinase adsorption on Streamline SP ion-exchange adsorbent, using a crude fermentative broth with cells as the feedstock. The kinetics and mass transfer parameters were estimated using the PSO (particle swarm optimization) heuristic algorithm. The parameters were estimated for each expansion degree (ED) using three breakthrough curves at initial inulinase concentrations of 65.6UmL(-1). In sequence, the model parameters for an ED of 2.5 were validated using the breakthrough curve at an initial concentration of 114.4UmL(-1). The applicability of the validated model in process optimization was investigated, using the model as a process simulator and experimental design methodology to optimize the column and process efficiencies. The results demonstrated the usefulness of this methodology for expanded bed adsorption processes.

Modeling of ion exchange expanded-bed chromatography for the purification of C-phycocyanin.

This work is focused on the experimental evaluation and mathematical modeling of ion exchange expanded-bed chromatography for the purification of C-phycocyanin from crude fermentative broth containing Spirulina platensis cells. Experiments were carried out in different expansion degree to evaluate the process performance. The experimental breakthrough curves were used to estimate the mass transfer and kinetics parameters of the proposed model, using the Particle Swarm Optimization algorithm (PSO). The proposed model satisfactorily fitted the experimental data. The results from the model application pointed out that the increase in the initial bed height does not influence the process efficiency, however enables the operation of expanded-bed column at high volumetric flow rates, improving the productivity. It was also shown that the use of mathematical modeling was a good and promising tool for the optimization of chromatographic processes.

Bioproduct Extraction From Microbial Cells by Conventional and Nonconventional Techniques

Abstract Demands of the food industry for sustainability have increased interest in microbial processes, which may yield bioproducts, such as pigments, lipids, proteins, biopolymers, enzymes, and carbohydrates, besides other bioactive molecules and ingredients from renewable feedstocks. The main focus of research and development has been the determination of cultivation parameters. However, intracellular bioproduct extraction from cells is a crucial step of the process, because it should aim at high yield, mitigation of environmental impacts, easy scale-up, economic feasibility, and integration in the downstream process. In this chapter, the main goals are to review conventional and nonconventional techniques involved in the physicochemical extraction of bioproducts from fungi, bacteria, and microalgae, and to describe their benefits and constraints. Techniques, such as supercritical fluid and ultrasound-assisted extraction are discussed, as well as emerging technologies, such as microwave-assisted extraction, pulsed electric fields, and organic solvent-free systems.

Cell pretreatment with ethylenediaminetetraacetic acid for selective extraction of C-phycocyanin with food grade purity

C‐phycocyanin (C‐PC) is a natural blue dye, and depending on its purity, which is measured by the ratio between the absorbance of the chromophore (A620) and the absorbance of the proteins (A280), it can be used in food (purity > 0.7), cosmetics (purity > 1.5), and therapeutic treatments (purity > 4.0). Several physical, chemical, and enzymatic methods of extraction are reported, however, few are able to extract C‐PC with purity above 0.7. An innovative method of C‐PC extraction with food grade purity from wet Spirulina platensis biomass is proposed. The cells were pretreated with ethylenediaminetetraacetic acid and subsequent C‐PC extraction was performed with tris‐(hydroxymethyl) aminomethane‐SO4 buffer. C‐PC was released after 12 h of cell pretreatment. Six variables of the extraction process were evaluated. The extraction temperature significantly influenced C‐PC extraction yield and purity. In the best condition of cell pretreatment and extraction, C‐PC with purity of 1.0 and extraction yield of 129.0 mg/g could be obtained to be used as a food dye without any purification process. Lastly, an ultrafiltration process was integrated and C‐PC was concentrated 8.8‐fold, resulting in purity of 1.6 and recovery of 93.4%.