INVESTIGADORES
WOLMAN Federico Javier
artículos
Título:
Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase
Autor/es:
LUCÍA VIRGINIA ROMERO; ALEXANDRA MARISA TARGOVNIK; FEDERICO JAVIER WOLMAN; OSVALDO CASCONE; MARÍA VICTORIA MIRANDA
Revista:
BIOTECHNOLOGY LETTERS
Editorial:
SPRINGER
Referencias:
Año: 2011 vol. 33 p. 947 - 956
ISSN:
0141-5492
Resumen:
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 27_C instead of at 24_C, 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 D
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.