Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase

ROMERO, LUCÍA VIRGINIA; TARGOVNIK, ALEXANDRA MARISA; WOLMAN, FEDERICO JAVIER; CASCONE, OSVALDO; MIRANDA, MARÍA VICTORIA

BIOTECHNOLOGY LETTERS

SPRINGER

Año: 2011

0141-5492

  A process based on orally infected Rachiplusia nu larvae as biological factories for expression and one-step purification of horseradish peroxidase isozyme C (HRPC) is described. The process allows obtaining high levels of pure HRPC by membrane chromatography purification. The introduction of the partial polyhedrin homology sequence (PPHS) element in the target gene increased 2.8 fold the HRPC expression level, whereas it increased 1.8 fold when the larvae were reared at 27° C instead of at 24° C, summing up a 4.6-fold overall increase in the expression level. Additionally, HRPC purification by membrane chromatography at a high flow rate greatly increases the productivity without affecting the resolution. The maximum velocity and Michaelis constant values of the recombinant HRPC are similar to those of the HRP from Armoracia rusticana roots.