Structural analysis of halogenases identified in biosynthetic gene clusters of glycopeptide antibiotics.

• Grant number: 12/23427-7
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2013
• End date: August 31, 2015
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Marcio Vinicius Bertacine Dias
• Grantee: Tábata Peres Cardoso
• 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
• Associated scholarship(s): 14/15510-7 - Investigation of the function and requirement of two halogenases from the glycopeptide A47934 biosynthetic gene cluster, BE.EP.MS

Abstract

Although the organohalogen products had been considered in the past as eccentric compounds, nowadays they represent an important role in the research of natural products. Various experiments have shown that natural compounds may have their biologic activity altered by presence of absence of halogen. Glycopeptide antibiotics, such as vancomycin and teicoplanin are halogenated natural products and they have substantial importance in the medicine. They are important molecules with clinical applications in the treatment of gram-positive infections, mainly against S. aureus, including meticilin resistant strains. It has been proved that the absence of a single chloride atom in the vancomycin molecule (dechlorovancomycin) causes the reduction of its activity in more than 70%. This observation shows the importance of the chloride atoms for this group of antibiotics. However, the halogenases, which are the enzymes responsible to catalyse the attachment of halogens in natural products, are not very understood. In this project, we intend to study two halogenases identified in the gene cluster for biosynthesis of glycopeptide antibiotics (teicoplanin and complestatin). We aims to establish protocols for production of these enzymes in soluble form, obtain crystals and solver their structures. The crystallographic structure of these enzymes may give crucial information about the active site which may be target of site direct mutagenesis to increase their promiscuity and consequently apply them to produce new derivatives of glycopeptides through enzymatic chemistry or combinatorial biosynthesis.

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)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

CARDOSO, Tábata Peres. Análise estrutural de halogenases encontradas em clusters gênicos da biossíntese de antibióticos glicopeptídicos. 2015. Master's Dissertation - Universidade Estadual Paulista (Unesp). Instituto de Biociências Letras e Ciências Exatas. São José do Rio Preto São José do Rio Preto.