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Simultaneous Cell Disruption and Aqueous Two-Phase Extraction for Isolation of Intracellular Recombinant Proteins

Chinese Journal of Chemical Engineering 1999年第2期7卷 139-144页

作者：王光虎冯小黎苏志国 laboratoryofbiochemicalengineering DalianUniversityofTechnologyDalian116012China Staekeylaboratoryofbiochemicalengineering InstituteofChemicalMetallurgyChineseAcademyofSciencesBeijing100080China

A new technique was developed for the integrated processing of cell disruption and aqueous two-phase extraction in a high-speed bead mill to separate intracellular proteins from microbial cells. The process was named as simultaneous cell disruption and aqueous two-phase extraction (SDATE). Advantages, such as high cell disruption efficiency, biochemical activities preservation of proteins, cell debris elimination, and prelimiary purification of the target protein were being claimed. When this technique was employed for isolating recombinant Tumor Necrosis Factor (TNF) from E. coli, overall protein concentration and TNF activity were found to have been increased. More than 95% of TNF was partitioned into the top phase and all cell debris were in the bottom phase. The partition coefficient was greater than 3 and the TNF purification factor was greater than 6. It is shown that less separation steps were being utilized in the new technique, meaning a reduction in separation time and less process extractors required.

关键词： SDATE release of intracellular proteins high speed bead mill aqueous two-phase extraction tumor necrosis factor

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