EXPRESSION OF THE LIGNIN PEROXIDASE H2 GENE FROM PHANEROCHAETE-CHRYSOSPORIUM IN ESCHERICHIA-COLI

The DNA sequence for the extracellular lignin peroxidase isozyme H2 fr om Phanerochaete chrysosporium, obtained from cDNA clone lambda ML-6, was synthesized by PCR and successfully expressed in Escherichia coli under control of the T7 promoter. The portion of the cDNA encoding the signal peptide, not found in the mature native enzyme, was not includ ed. Recombinated lignin peroxidase H2 (rLiPH2) was produced in inclusi on bodies in an inactive form. Active enzyme was obtained by refolding with glutathione-mediated oxidation in a medium containing urea, Ca2, and hemin. The recombinant enzyme had spectral characteristics and k inetic properties identical to that of native enzyme isolated from P. chrysosporium. Surprisingly, rLiPH2, like the native enzyme, also exhi bited some manganese peroxidase activity. (C) 1998 Academic Press.