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Cloning, expression, purification and spectrophotometric analysis of lanosterol 14-alpha demethylase from Leishmania braziliensis (LbCYP51)

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Abstract

Leishmaniasis, a neglected tropical disease, is a major cause of morbidity and mortality worldwide. Of the three main clinical forms, cutaneous leishmaniasis (CL) is the most common and 40 million people are at risk in the endemic areas. Currently, the available drugs to fight leishmaniasis have high toxicity and poor efficiency. Then, it is very important to search for effective and safe drugs that would target essential enzymes from the parasite, such as lanosterol 14-alpha demethylase (CYP51, EC 1.14.13.70) from Leishmania braziliensis. Because most drug design efforts have been directed for Leishmania non-braziliensis species, there is no structural or kinetic data regarding L. braziliensis CYP51. Herein, we present for the first time molecular biology efforts and purification protocol to obtain the enzyme LbCYP51. These results lay the ground for future investigation of drugs against this target.

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Funding

Funding was provided by fapesb (Grant No. PNE009/2011).

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Authors and Affiliations

  1. Faculdade de Farmácia, Universidade Federal da Bahia, Salvador, BA, 40170-290, Brazil

    Humberto F. Freitas, Acássia Benjamim Leal Pires & Marcelo S. Castilho

  2. Programa de pós-graduação em Biotecnologia, Universidade Estadual de Feira de Santana, Feira de Santana, BA, Brazil

    Humberto F. Freitas & Marcelo S. Castilho

  3. Departamento de Ciências da Vida, Universidade do Estado da Bahia, Salvador, BA, 41150-000, Brazil

    Acássia Benjamim Leal Pires

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  1. Humberto F. Freitas
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Correspondence to Marcelo S. Castilho.

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Freitas, H.F., Leal Pires, A.B. & Castilho, M.S. Cloning, expression, purification and spectrophotometric analysis of lanosterol 14-alpha demethylase from Leishmania braziliensis (LbCYP51). Mol Biol Rep 45, 175–183 (2018). https://doi.org/10.1007/s11033-018-4150-7

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