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標題: Cell disruption enhanced the pure EGFP recovery from an EGFP-intein-surface protein production system in recombinant E. coli
作者: Wu, Jiun-Yan
Chen, Chih-I
Chen, Chi-Ming
Lina, Chia-Chi
Kana, Shu-Chen
Shieh, Chwen-Jen
Liua, Yung-Chuan
關鍵字: Protein Purification;Surface display;Intein;Centrifugation;Recombinant DNA
Project: Biochemical Engineering Journal, Volume 68, page(s) 12– 18.
In this study, three approaches for protein production were devised to enhance the efficiencies in yielding
high purity protein via surface display system. A plasmid carrying enhanced green fluorescent protein
(EGFP), an intein (INT) and ice nucleation protein (INP) was constructed to produce EGFP via surface
display in Escherichia coli. To obtain high purity of the produced EGFP, several procedures, including
osmotic shock, surfactant addition and cell disruption were employed. Among these approaches, the
cell disruption method gave the highest EGFP purity by simply conducting several centrifugations. An
EGFP yield of 63 mg/L with 97% purity was obtained. The result demonstrated that pure EGFP can be
harvested only through centrifugation; no complicated processes or expensive equipment are required.
This approach shows potential for the production of pure recombinant proteins in scale-up processes for
biotechnological, academic and industrial uses.
ISSN: 1369-703X
DOI: 10.1016/j.bej.2012.06.020
Appears in Collections:化學工程學系所

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