Obtainment and crystallization of glycosyltransferases of biotechnological interest of the macrolide antibiotic biosynthesis

• Grant number: 12/16631-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2012
• End date: October 31, 2013
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Marcio Vinicius Bertacine Dias
• Grantee: Larissa Antelo de Sá
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 10/15971-3 - Structural characterization of enzymes from antibiotic biosynthetic pathways with biotechnological interest, AP.JP

Abstract

Natural products represent one of the principal sources of bioactive molecules which can be used therapeutically on humans. Within the natural products, various antibiotics exist which have been utilized throughout many decades against various types of infections. Natural antibiotics are chemically classified by their action mechanism and they are generally complex molecules produced by micro-organisms, plants, and marine organisms whose chemical manipulation is not simple. These organisms produce bioactive molecules through many enzymatic pathways that can be explored biotechnologically by the production of known antibiotics derivates. Meanwhile, for this, the determination of its crystallographic structures is necessary. Among the enzymes related to biosynthesis, the glycosyltransferases are of great importance, since almost every natural product has sugars bonded to the aglycone. In the biosynthesis of macrolide antibiotics, some glycosyltransferases are peculiar given that they form a complex with a homolog accessory enzyme on the P450 cytochrome, whose function is not well established. In this project, which is part of a young research project that aims to study, structurally, various enzymes of biotechnological importance on the production of antibiotics, we intend to obtain glicosiltransferases related to the biosynthesis of macrolide antibiotics, more specifically the tylosin and puromycin. For this, the genes will be cloned, expressed and the proteins will be purified and crystallization trials will be performed to obtain crystals and consequently obtain the structure of these enzymes.(AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CARDOSO, TABATA P.; DE SA, LARISSA A.; BURY, PRISCILA DOS S.; CHAVEZ-PACHECO, SAIR M.; DIAS, MARCIO V. B.. Cloning, expression, purification and biophysical analysis of two putative halogenases from the glycopeptide A47,934 gene cluster of Streptomyces toyocaensis. Protein Expression and Purification, v. 132, p. 9-18, APR 2017. (10/15971-3, 12/23427-7, 12/16631-7)