   5.1.3.31 | recombinant enzyme from Escherichia coli inclusion bodies by treatment with 20 ml 10 mM Tris-HCl buffer, pH 7.5, containing 1.0 mM EDTA, 4% Triton X-100, incubation at room temperature for 30 min, twice. Then the inclusion bodies are treated with denaturant solution containing 50 mM Tris-HCl buffer, pH 7.5, containing 6 M guanidine-HCl, 0.2 M NaCl, 1 mM EDTA overnight at 4°C. The solubilized enzyme (100 mg protein in 100 ml solution) is gently dropped into 1 l of refolding buffer containing 0.1 M Tris-HCl, pH 7.5, 1 mM MnCl2, and 0.4 M L-arginine, and incubated for 16 h at 4°C, followed by ultrafiltration |
746648 |
