Recovery of Cytochrome c using Ionic Liquid-based Aqueous Two Phase System

• Main Authors: Yu-Yu Lin, 林宥佑
• Other Authors: Chi-Wei Lan
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/f7ax2b

碩士 === 元智大學 === 化學工程與材料科學學系 === 107 === Aqueous two phase system (ATPS) has gained an interest because of great potential for extraction, separation and purification in industry and academia. In recent years, ionic liquids (ILs)-based ATPS have attracted much attention due to their “green” characteristics such as the excellent thermal and electrochemical stability, non-flammability, chemical stability, strong solubility power, and negligible volatility. Cytochrome c is one of the cytochromes and is widely found in various animals, plants and microorganisms. Cytochrome c is an essential component of the electron transport chain, where it carries one electron. The medical application of cytochrome c is a cellular respiratory activator. The conventional steps of cytochrome c extraction from tissue were complicated. Accordingly, the objective of this study is to optimize the condition of ionic liquid based aqueous two phase system for cytochrome C recovery. Firstly, the partition behavior of cytochrome c standard in the [C6mim] Br/citrate ATPS was studied. The effects of system composition, pH, temperature and addition of sodium chloride upon recovery of cytochrome c using IL-based ATPS were investigated and discussed. Secondly, culture broth containing cytochrome c standard solution will be used to simulate cell lysis and release cytochrome c to investigate the recovery of cytochrome c in IL-based ATPS. The effects of system composition, pH, temperature, and sodium chloride addition upon recovery of cytochrome c will be investigated and discussed. The results of the first part indicated that the optimal conditions for cytochrome c recovery were obtained with IL-based ATPS consisted 28% (w/w) [C6mim] Br and 22% (w/w) citrate at pH 9 and room temperature (25℃) with addition of 0.6% (w/w) sodium chloride. The partition coefficient was 220.20 ± 3.82. Furthermore, the results of the second part showed that the optimal conditions of cytochrome c recovery in mixture model were obtained with IL-based ATPS consisted 28% (w/w) [C6mim] Br and 24% (w/w) citrate at pH 9 and room temperature (25℃) with addition of 0.2% (w/w) sodium chloride. The efficiency of recovery achieved 99.76%. In summary, IL-based ATPS is a promising technique to recover cytochrome c in one-step operation.