Recombinant peroxidase production in species of lepidoptera frequently found in Argentina

LUCÍA ROMERO; ALEXANDRA TARGOVNIK; FEDERICO WOLMAN; MARIELA FOGAR; MARÍA SIMONELLA; OSVALDO CASCONE; MARÍA MIRANDA

New Biotechnology

Elsevier

Lugar: Cambridge; Año: 2010 vol. 27 p. 857 - 861

1871-6784

Abstract: 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 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 ìg per larva. Rachiplusia nu was highly susceptible to oral infection, reaching 22 ìg 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.