The Extraction Separation of Lipase by Triblock Copolymer Aqueous Two-Phase System

• Main Authors: Wu, Ming-chien, 吳明鍵
• Other Authors: Tsay Sun-Yuan
• Format: Others
• Language: zh-TW
• Published: 1998
• Online Access: http://ndltd.ncl.edu.tw/handle/31334374039702628844

碩士 === 國立成功大學 === 化學工程學系 === 86 === It is important to maintain the activity of enzyme and reduce the cost of purification. In this study, enzymes such as C. rugosa lipase and A. radioresistens lipase were extracted by using aqueous two-phase extraction system. It was carried out by using triblock copolymer as one phase of the aqueous two-phase system. High selectivity and high productivity can be theadvantages from the aqueous two-phase extraction system. Acrylic polyampholyteshave isoelectric points at which the polyampholytes can precipitate.Hence, it can be recycled after use. The components of the two-phase systemare triblock copolymer and another aqueous solution such as dextran or PEG solution. Both B16M12A8 and PEG20000 solution have shown influence onthe activity of C. rugosa lipase. The two-phase system was made of 20%w/v PEG20000 with 8% w/v B16M12A8, 20% w/v PEG20000 with 3% w/v B36M36A36,and 20% w/v dextran40000 with 3% w/v B36M36A36, etc. The effect of salts such as KCl, NaCl, and FeCl2 on the separation of the enzyme was also discussed.The concentrations of the salts were 0.03M,0.06M, and 0.09M, respectively.It is also important to discuss the effect of the temperature of the two-phasesystem on the separation of the enzymes. In addition, the triblock copolymer was precipitated from the aqueous two-phase system so that it can be recycled.In this way, the enzyme can also be recovered from this separation system.The activity of the lipase in the liquid phase after centrifuge and in thedissolved precipitate was measured.