Single-step purification of the recombinant green fluorescent protein from intact Escherichia coli cells using preparative PAGE.
Mechanical and non-mechanical breakages of bacterial cells are usually the preliminary steps in intracellular protein purification. In this study, the recombinant green fluorescent protein (GFP) was purified from intact Escherichia coli cells using preparative PAGE. In this purification process, cells disruption step is not needed. The cellular content of E. coli was drifted out electrically from cells and the negatively charged GFP was further electroeluted from polyacrylamide gel column. SEM investigation of the electrophoresed cells revealed substantial structural damage at the cellular level. This integrated purification technique has successfully recovered the intracellular GFP with a yield of 82% and purity of 95%.
Chew
Few Ne
FN
Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia, Selangor, Malaysia.
Tan
Wen Siang
WS
Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Selangor, Malaysia.|Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.
Ling
Tau Chuan
TC
Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, Selangor, Malaysia.
Tey
Beng Ti
BT
Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia, Selangor, Malaysia.|Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.
D018533:Biomass; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000302:isolation & purification*; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D011994:Recombinant Proteins / Q000302:isolation & purification*
D011994:Recombinant Proteins; C485192:green fluorescent protein, Aequorea victoria; D049452:Green Fluorescent Proteins
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