Induction, characterization, and heterologous expression of a carotenoid degrading versatile peroxidase from Pleurotus sapidus

Abstract

Abstract A β-carotene degrading versatile peroxidase (VP) was successfully induced in submerged cultures of the white-rot basidiomycete Pleurotus sapidus by use of residues of a biogas plant as a carbon and nitrogen source. After three chromatographic steps, the VP was isolated with an overall yield of 12% and a purification factor of 130. The purified enzyme showed a molecular mass of 38 kDa and an isoelectric point of 3.6. Highest affinity to β-carotene ( K m = 50 ± 5 μM) was observed at 30 °C and pH 4.5. The purified VP was capable of degrading suspended lignin organosolv particles. N-terminal and internal peptide sequences were obtained from Edman degradation and mass spectrometric peptide sequencing. The VP encoding cDNA was identified by colony hybridization and amplified by PCR. Bioinformatic analyses revealed an open reading frame of 1083 bp and similarities of 90% to VPs from P. eryngii . The recombinant VP was produced successfully with an activity of 450 ± 20 mU mg −1 in cultures of H. polymorpha .