Purification of green fluorescent protein overexpressed by a mutant recombinant Escherichia coli

Green fluorescent protein was purified from sonicated recombinant Escherichia coli and its mutant obtained after exposure to UV light. The latter overexpresses green fluorescent protein. The two-step procedure consisted of a two-phase aqueous extraction with PEG/salt and precipitation of the proteins from PEG phase by free Zn2+. The recoveries of green fluorescent protein were 73 and 83% in the cases of recombinant E. coli and its mutant, respectively. The corresponding fold purifications were 24 and 9, respectively. In both cases, the purified protein showed a single band on SDS-PAGE corresponding to 28 kDa.