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<Emphasis Type="Italic">Rachiplusia nu</Emphasis> larva as a biofactory to achieve high level expression of horseradish peroxidase
Authors:Lucía Virginia Romero  Alexandra Marisa Targovnik  Federico Javier Wolman  Osvaldo Cascone  María Victoria Miranda
Institution:1.Cátedra de Microbiología Industrial y Biotecnología,Facultad de Farmacia y Bioquímica (UBA),Ciudad Autónoma de Buenos Aires,Argentina
Abstract: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.
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