Study of the friedelin production from the gene mutations of friedelin synthase of Maytenus ilicifolia

• Grant number: 12/21005-8
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2013
• End date: April 30, 2014
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Organic Chemistry
• Principal Investigator: Maysa Furlan
• Grantee: Karina Alves Pinheiro
• Host Institution: Instituto de Química (IQ). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

In previous work, the phytochemical study of the root bark of seedelings of Maytenus ilicifolia (Celastraceae), associated with the evaluation of biological activity, led to the isolation of quninemethide triterpenes with potent antioxidant and antitumor activities. Among the isolated quinonemethides, those who showed the best biological results were maytenin, 22 ²-hydroximaytenin and pristimerin, which are considered minority metabolites in such species. Phytochemical studies comparing young and adult plants of Maytenus ilicifolia also indicated that the quninonemethide triterpenes are accumulated in the young plant, while the adult plant produces pyridine alkaloids sesquiterpenes as the major metabolites. In this context, the biosynthetic studies of the quinonemethide triterpenes were performed using as precursor the 2,3-epoxiesqualeno in the presence of enzymatic extract from the root bark, leaves and stems of young plants (seedlings) of M. ilicifolia, which proved to be the friedelin the key intermediate in the biosynthesis thereof. The biosynthetic studies allowed introduced a vast knowledge of the conditions of extraction, analysis, characterization and compartmentalization of the enzyme and proteomic studies and regulation of this pathway are being developed by our research group. Therefore, this project aims to explore the production of friedelin more specifically by the action of the enzyme oxidosqualene cyclase (friedelin synthase), using a heterologous expression in Saccharomyces cerevisiae gene encoding this enzyme in M. ilicifolia. The cloning of oxidosqualene cyclase of the leaves of M. ilicifolia has already been done by our group, as well as expression studies in S. cerevisiae genetically modified for production of triterpenes. These studies can be supplemented by determining the specificity of the friedelin production mechanism through site-directed mutations of the gene that encoding the friedelin synthase which is expressed in S. cerevisiae in order to elucidated their production. The production of friedelin and other metabolites in S. cerevisiae will be assessed by chromatographic methods to obtain the best production profiles of triterpene of interest. (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)

SOUZA-MOREIRA, TATIANA M.; ALVES, THAS B.; PINHEIRO, KARINA A.; FELIPPE, LIDIANE G.; DE LIMA, GUSTAVO M. A.; WATANABE, TATIANA F.; BARBOSA, CRISTINA C.; SANTOS, VANIA A. F. F. M.; LOPES, NORBERTO P.; VALENTINI, SANDRO R.; et al. Friedelin Synthase from Maytenus ilicifolia: Leucine 482 Plays an Essential Role in the Production of the Most Rearranged Pentacyclic Triterpene. SCIENTIFIC REPORTS, v. 6, NOV 22 2016. (12/21005-8, 13/07600-3, 13/15306-8, 14/03819-3, 11/10379-1, 12/09584-2)

Academic Publications

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

PINHEIRO, Karina Alves. Estudo da formação da friedelina a partir de mutações no gene da friedelina sintase. 2015. Master's Dissertation - Universidade Estadual Paulista (Unesp). Instituto de Química. Araraquara Araraquara.