Recombinant peroxidase production in species of lepidoptera frequently found in Argentina

LUCIA ROMERO; ALEXANDRA TARGOVNIK; FEDERICO WOLMAN; M. FOGAR; M. SIMONELLA; O. CASCONE; M.V. MIRANDA

NEW BIOTECHNOLOGY

ELSEVIER SCIENCE BV

Año: 2010

1871-6784

Horseradish peroxidase isozyme C (HRPC) is an important commercial biocatalyst. In this study, a screening of different lepidopteran species frequently found in Argentina to produce this protein was carried out. Two recombinant viruses were constructed: AcMNPV HRPC polyhedrin-minus (occ-), an intrahemocelical infective virus; and AcMNPV HRPC polyhedrin-plus (occ+), to achieve an oral infective baculovirus. Each lepidopteran species was infected either with AcMNPV HRPC occ- or AcMNPV HRPC occ+ and the harvesting time post-infection (dpi) were optimized. All species were susceptible to AcMNPV HRPC occ- infection, giving Spodoptera frugiperda the best yield: 41 µg per larva. Rachiplusia nu was highly susceptible to oral infection, reaching 22 µg per larva at 4 dpi. HRPC was purified by IMAC from S. frugiperda extracts with a yield of 86% and a purification factor of 29.