Resumo

PLIEGO-ARREAGA, R.; REGALADO, C.; AMARO-REYES, A.  e  GARCIA-ALMENDAREZ, B.E.. Lactose-induced expression of recombinant turnip peroxidase in Escherichia coli. Rev. Mex. Ing. Quím [online]. 2013, vol.12, n.3, pp.505-511. ISSN 1665-2738.

Turnip (Brassica napus L. var. purple top white globe) peroxidases (E.C. 1.11.1.7) may be applied free or immobilized on phenolic compounds degradation in waste water, in addition to its use in diagnostic test kits. The aim of this work was to achieve heterologous expression by lactose-induction of one peroxidase isoform (PodC) using Escherichia coli Rosetta 2 to produce recombinant turnip peroxidase in a fermenter. Recombinant PodC contained in inclusion bodies was refolded under oxidative conditions and purified by IMAC with a yield of 36 mg L^-1 of culture. The enzyme showed a specific activity of 1004 ABTS units mg^-1 under optimum conditions (pH 6, 45 °C). The value of K_M for ABTS was 0. 38 mM and the catalytic efficiency (k_cat/K_M) was 5.4×10⁶ s^-1 M^-1. The use of lactose as inducer and scale up of the fermentation process may effectively decrease the production cost of recombinant PodC, which in turn can be applied for biotechnological applications.

Palavras-chave : Brassica napus; peroxidase; expression; lactose; fermenter.

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