碩士 / 明志科技大學 / 生化工程研究所 / 100 / In this study, the adsorption conditions for C-phycocyanin (C-PC) from cyanobacteria Spirulina platensin in small packed beds were optimized by using design of experiments. The optimal adsorption conditions were further applied to expanded bed and stirred fluidized bed adsorption processes for purification of C-PC.
The choice of cell disruption method (i.e., glass bead mill and ultrasonication) and the stability of C-PC (e.g., adsorption pH, temperature, and light) were investigated in preliminary experiments. The results showed that the release of C-PC from the disrupted cells by using ultrasonication was higher than that by using high speed glass bead mill. The C-PC solutions showed a slow rate of degradation in the dark at the temperature of 4-25 ℃, however, the degradation rate increased dramatically in white light at temperature of 25℃. Moreover, the C-PC solution was highly stable at pH 6-8.
Before running the design of experiments, STREAMLINE DEAE and Phenyl adsorbents were firstly employed to evaluate the adsorption performance under the different conditions. It was observed that the ion exchange DEAE adsorbent has higher capture for C-PC at pH of 6-8, however, hydrophobic Phenyl adsorbent has higher capture for C-PC at ammonium sulfate concentration of 0.5-1.0 M. As compare to the binding capacity for C-PC by using these two different adsorbents, the ion exchange adsorbent has higher capture ability than hydrophobic adsorbent at the same pH value. Hence, the STREAMLINE DEAE ion exchange adsorbent was chosen to operate the further design of experiments in packed bed (i.d. 1.6 cm × 40 cm)
The design of experiments was operated with factorial design and response surface techniques. The influences of adsorption pH, operating flow rate, algal concentration, and packed bed height on the dynamic binding capacity for C-PC were evaluated to determine the optimal adsorption conditions for C-PC at 5% breakthrough in packed bed. The results showed that the optimal conditions for capture C-PC were pH 7.0, flow rate 10 mL/min, algal concentration 10% (w/v), and packed bed height 1.6 cm. Moreover, the optimal condition in the elution stage was found to be 0.6 M NaCl (pH 7) and flow rate 2.6 mL/min.
The optimal adsorption and elution conditions obtained from the design of experiments in packed beds could straightforwardly be applied to the expanded bed and stirred fluidized bed adsorption processes for directly purifying the C-PC from Spirulina platensin. The results showed that the purification of C-PC by using expanded bed and stirred fluidized bed chromatography was recovered with a yield of 47.70 % and 59.42% with a purification factor 2.7 and 3.00, respectively. Therefore, the stirred fluidized bed chromatography would be more suitable for purification of target protein in high biomass systems.
| Identifer | oai:union.ndltd.org:TW/100MIT00723005 |
| Date | January 2012 |
| Creators | Hsu, Shihlung, 徐仕龍 |
| Contributors | Chang, Yukaung, 張煜光 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 98 |
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