| Summary: | 碩士 === 中原大學 === 化學工程研究所 === 103 === In recent years, the trend has been developed rapid, efficient and scale up for separation and purification of protein . Three-phase partitioning ,a protein purification approach, is carried out by mixing ammonium sulfate,buffer and organic solvents to obtain partitioned organic phase, interfacial precipitate and aqueous phase. The effects of different salt concentration, organic solvents, and pHs of various protein has been the focus of investigation. The purpose of this thesis is to apply three-phase partitioning (TPP) to obtain correctly refolded active proteins from solubilized inclusion bodies. In this study, the recovery recombinant streptavidin, an important bio-conjugation protein for various biotechnology applications, were isolated from inclusion bodies by using three-phase partitioning (TPP). The system consisted of crude enzyme extract : t-butanol 1:1 (w/v),5% ammonia sulfate, pH 8.0(Tris Buffer) with incubation temperature of 37 oC showed the highest protein purification(fold) (3.67) and yield(%) (164.5%). In conclusion , protocol of streptavidin purification can be further developed to Streptavidin ,which can see wide applications in biosensing and preparative biotechnology field .
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