earticle

영어

cDNA sequence encoding CWPO_C from Populus alba L was heterologously expressed in E. coli as inclusion bodies. Insoluble products were solubilized and reactivated via refolding procedure. The efficiency of refolding was estimated throughout oxidation rate of ABTS. The optimal conditions for refolding CWPO_C with systematically optimized parameters are: 100 mM Tris-HCl at pH 8.5, 0.6 mM GSSG, 5 μM hemin, 0.6 M GmdCl and 5mM CaCl2. Property of substrate preference to sinapyl alcohol rather than coniferyl alcohol was characterized with refolded CWPO_C. This unique property of refolded CWPO_C was confirmed, like native CWPO_C when relative oxidation rate of these monolignols catalyzed by refolded CWPO_C and HRP-C were compared in this study. Successful expression of CWPO_C in E. coli provides a valuable tool for elucidate the structure - function relationship of CWPO_C known as a S-peroxidase having important role in the lignification of angiosperm woody plant cell walls.