Structural analysis of key enzymes for gentamicin and sisomicin biosynthesis

• Grant number: 14/07843-6
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: November 01, 2014
• End date: September 30, 2019
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Marcio Vinicius Bertacine Dias
• Grantee: Priscila dos Santos Bury
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 10/15971-3 - Structural characterization of enzymes from antibiotic biosynthetic pathways with biotechnological interest, AP.JP

Abstract

Aminoglycosides or aminocyclitols are molecules which have a core and several other unusual sugars, such as aminosugars and deoxysugars attached. All these structures and their functional groups are essential for the antibiotic activity, which is the inhibition of the protein synthesis because its interaction with the bacterial ribosome 30S subunit. Aminoglycoside antibiotics are molecules extensively used in the medicine, despite their toxic effects, for the treatment of infectious diseases. Also as most of natural products of microbial origin, aminoglycosides are secondary metabolism products of bacteria and the genes responsible for its biosynthesis are assembled in gene clusters. Various gene clusters for biosynthetic pathways of aminoglycosides in different organisms have been identified and sequenced. However, not much has been done with the corresponding enzymes, mainly those ones for the biosynthesis of gentamicin, one of the aminoglycosides with ample use in the human medicine. Gentamicin is a 4,6-DOS disubstituted and has two aminosugars, garosamine and purporosamine. Sisomicin is another aminoglycoside which have high similarity to the gentamicin and the unique difference is an ethylene bound in the purpurosamine group. Although the important of these aminoglycosides and the knowledge of their biosynthetic pathway, very little is known about their enzymes, mainly about their crystallographic structures and consequently there is a limitation to explore these enzymes in biotechnological process of combinatorial biosynthesis to produce new derivatives of aminoglycosides. In this project we intend to perform a structural study of key enzymes of the biosynthesis of gentamicin and sisomicin that have biotechnological interest and which there is not any information about their structures. Thus, the genes for those enzymes will be cloned, the proteins will be expressed and crystallization trials will be performed. We hope to obtain crystal and consequently solve their crystallography structures. In collaboration with other research groups, experiments of functional characterization will be done. In the end, the combination of structural and functional results for those enzymes might contribute to understand the aminoglycoside biosynthesis and also reveal crucial information that might be useful in biotechnological processes to produce new derivatives of aminoglycosides with lower toxicity. (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)

DE ARAUJO, NATALIA CERRONE; BURY, PRISCILA DOS SANTOS; TAVARES, MAURICIO TEMOTHEO; HUANG, FANGLU; PARISE-FILHO, ROBERTO; LEADLAY, PETER; BERTACINE DIAS, MARCIO VINICIUS. Crystal Structure of GenD2, an NAD-Dependent Oxidoreductase Involved in the Biosynthesis of Gentamicin. ACS Chemical Biology, v. 14, n. 5, p. 925-933, MAY 2019. (10/15971-3, 15/09188-8, 14/07843-6, 14/50324-0)

BURY, PRISCILA DOS SANTOS; HUANG, FANGLU; LI, SICONG; SUN, YUHUI; LEADLAY, PETER F.; BERTACINE DIAS, MARCIO VINICIUS. Structural Basis of the Selectivity of GenN, an Aminoglycoside N-Methyltransferase Involved in Gentamicin Biosynthesis. ACS Chemical Biology, v. 12, n. 11, p. 2779-2787, NOV 2017. (14/07843-6, 10/15971-3, 15/09188-8, 14/50324-0)

LI, SICONG; BURY, PRISCILA DOS SANTOS; HUANG, FANGLU; GUO, JUNHONG; SUN, GUO; REVA, ANNA; HUANG, CHUAN; JIAN, XINYUN; LI, YUAN; ZHOU, JIAHAI; et al. Mechanistic Insights into Dideoxygenation in Gentamicin Biosynthesis. ACS CATALYSIS, v. 11, n. 19, p. 12274-12283, OCT 1 2021. (14/07843-6, 15/09188-8, 10/15971-3, 18/00351-1)

Academic Publications

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

BURY, Priscila dos Santos. Structural analysis of key enzymes for gentamicin and sisomycin biosynthesis.. 2019. Doctoral Thesis - Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) São Paulo.