2.1.1.251 | recombinant enzyme from inclusion bodies harvested from cells solubilized using 6 M guanidine hydrochloride, 50 mM DTT, and 20 mM 2-mercaptoethanol. The solubilized protein is diluted 60fold with protein refolding at room temperature for 24 h in refolding buffer consisting of 500 mM Tris, pH 8.0, 1 M KCl, 10 mM DTT, 10 mM 2-mercaptoethanol, and 100 mM ZnCl2. Refolding buffers containing 500 mM guanidine hydrochloride, 33 mM CHAPS, 0.5% Triton X-100, 5 mM SDS, or 20% glycerol do not produce active methyltransferase, nor do refolding at 4°C in the optimal refolding buffer |
717763 |