MIRANDA Maria Victoria
Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase.
ROMERO, L; TARGOVNIK A; WOLMAN F; CASCONE O; MIRANDA MARÍA VICTORIA
Año: 2011 p. 947 - 947
A process based on orally-infected Rachiplusia nu larvae as biological factories for expression and one-step purification of horseradish peroxidase isozyme C(HRP-C) is described. The process allows obtaining high levels of pure HRP-C by membrane chromatography purification. The introduction of the partial polyhedrin homology sequence element in the target gene increased HRP-C expression level by 2.8-fold whereas it increased 1.8-fold when the larvae were reared at 27ºC instead of at 24ºC, summing up a 4.6-fold overall increase in the expression level. Additionally, HRP-C purification by membrane chromatography at a high flow rate greatly increase the productivity without affecting the resolution. The Vmax and Km values of the recombinant HRP-C were similar to those of the HRP from Armoracia rusticana roots.