Autor(es): Mazzeu, Bruna F. [UNESP] ; Souza-Moreira, Tatiana M. [UNESP] ; Oliveira, Andrew A. ; Remlinger, Melissa [UNESP] ; Felippe, Lidiane G. [UNESP] ; Valentini, Sandro R. [UNESP] ; Guido, Rafael V. C. ; Zanelli, Cleslei F. [UNESP] ; Furlan, Maysa [UNESP]
Data: 2022
Identificador Persistente: http://hdl.handle.net/11449/231554
Origem: Oasisbr
Assunto(s): Celastraceae; Friedelin synthase; Leu552Phe; Maytenus ilicifolia; Met549Ser; Saccharomyces cerevisiae; Site-directed mutation
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Friedelin, a pentacyclic triterpene found in the leaves of the Celastraceae species, demon-strates numerous biological activities and is a precursor of quinonemethide triterpenes, which are promising antitumoral agents. Friedelin is biosynthesized from the cyclization of 2,3-oxidosqualene, involving a series of rearrangements to form a ketone by deprotonation of the hydroxylated inter-mediate, without the aid of an oxidoreductase enzyme. Mutagenesis studies among oxidosqualene cyclases (OSCs) have demonstrated the influence of amino acid residues on rearrangements during substrate cyclization: loss of catalytic activity, stabilization, rearrangement control or specificity changing. In the present study, friedelin synthase from Maytenus ilicifolia (Celastraceae) was ex-pressed heterologously in Saccharomyces cerevisiae. Site-directed mutagenesis studies were performed by replacing phenylalanine with tryptophan at position 473 (Phe473Trp), methionine with serine at position 549 (Met549Ser) and leucine with phenylalanine at position 552 (Leu552Phe). Mutation Phe473Trp led to a total loss of function; mutants Met549Ser and Leu552Phe interfered with the enzyme specificity leading to enhanced friedelin production, in addition to α-amyrin and β-amyrin. Hence, these data showed that methionine 549 and leucine 552 are important residues for the function of this synthase.
Instituto de Química Universidade Estadual Paulista-UNESP, CP 355
Centro de Pesquisa e Inovação em Biodiversidade e Fármacos Instituto de Física de São Carlos Universidade de São Paulo
Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista-UNESP, Rod. Araraquara-Jaú km 1
Instituto de Química Universidade Estadual Paulista-UNESP, CP 355
Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista-UNESP, Rod. Araraquara-Jaú km 1
CAPES: 001
FAPESP: 2011/10379-1
FAPESP: 2013/07600-3
FAPESP: 2014/03819-3
FAPESP: 2014/25705-0
FAPESP: 2014/50926-0
