title: Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase.
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 V(max) and K(m) values of the recombinant HRP-C were similar to those of the HRP from Armoracia rusticana roots.
authors.affiliation: Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica (UBA), Junín 956 (1113), Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Lucía Virginia
authors.initials: LV
authors.lastname: Romero
authors.forename: Alexandra Marisa
authors.initials: AM
authors.lastname: Targovnik
authors.forename: Federico Javier
authors.initials: FJ
authors.lastname: Wolman
authors.forename: Osvaldo
authors.initials: O
authors.lastname: Cascone
authors.forename: María Victoria
authors.initials: MV
authors.lastname: Miranda
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods; D002845:Chromatography / Q000379:methods; D006735:Horseradish Peroxidase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007313:Insecta / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D007814:Larva / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D006735:Horseradish Peroxidase