Characterization and crystallization of recombinant pea cytosolic ascorbate peroxidase

摘要: An Escherichia coli expression system has been developed for pea cytosolic ascorbate peroxidase (APX). The enzyme was expressed as a fusion product with the E. maltose-binding protein rapid, affinity chromatography purification. Recombinant (rAPX) purified by tryptic digestion to separate from rAPX followed three chromatographic steps. demonstrated identical electrophoretic, enzymatic, and spectral properties when compared native APX isolated shoots. Upon addition of an equal molar amount H2O2, exhibits initial decrease in Soret maximum, which slowly converts stable, red-shifted peak similar that observed cytochrome c Compound I, indicating I consists oxyferryl (Fe(4+)-O) center. crystallized form suitable crystal structure determination, preliminary set data 2.6 A have collected.