The Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity

Friedelin, a pentacyclic triterpene found in the leaves of the Celastraceae species, demonstrates 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...

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Autores principales:	Bruna F. Mazzeu, Tatiana M. Souza-Moreira, Andrew A. Oliveira, Melissa Remlinger, Lidiane G. Felippe, Sandro R. Valentini, Rafael V. C. Guido, Cleslei F. Zanelli, Maysa Furlan
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Maytenus ilicifolia</i> Celastraceae <i>Saccharomyces cerevisiae</i> friedelin synthase site-directed mutation Met549Ser Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/ddc35de5e5134f5c9b696820b9fe8270
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spelling	oai:doaj.org-article:ddc35de5e5134f5c9b696820b9fe82702021-11-25T18:27:23ZThe Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity10.3390/molecules262268061420-3049https://doaj.org/article/ddc35de5e5134f5c9b696820b9fe82702021-11-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/22/6806https://doaj.org/toc/1420-3049Friedelin, a pentacyclic triterpene found in the leaves of the Celastraceae species, demonstrates 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 intermediate, 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 <i>Maytenus ilicifolia</i> (Celastraceae) was expressed heterologously in <i>Saccharomyces cerevisiae</i>. 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.Bruna F. MazzeuTatiana M. Souza-MoreiraAndrew A. OliveiraMelissa RemlingerLidiane G. FelippeSandro R. ValentiniRafael V. C. GuidoCleslei F. ZanelliMaysa FurlanMDPI AGarticle<i>Maytenus ilicifolia</i>Celastraceae<i>Saccharomyces cerevisiae</i>friedelin synthasesite-directed mutationMet549SerOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6806, p 6806 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Maytenus ilicifolia</i> Celastraceae <i>Saccharomyces cerevisiae</i> friedelin synthase site-directed mutation Met549Ser Organic chemistry QD241-441
spellingShingle	<i>Maytenus ilicifolia</i> Celastraceae <i>Saccharomyces cerevisiae</i> friedelin synthase site-directed mutation Met549Ser Organic chemistry QD241-441 Bruna F. Mazzeu Tatiana M. Souza-Moreira Andrew A. Oliveira Melissa Remlinger Lidiane G. Felippe Sandro R. Valentini Rafael V. C. Guido Cleslei F. Zanelli Maysa Furlan The Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity
description	Friedelin, a pentacyclic triterpene found in the leaves of the Celastraceae species, demonstrates 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 intermediate, 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 <i>Maytenus ilicifolia</i> (Celastraceae) was expressed heterologously in <i>Saccharomyces cerevisiae</i>. 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.
format	article
author	Bruna F. Mazzeu Tatiana M. Souza-Moreira Andrew A. Oliveira Melissa Remlinger Lidiane G. Felippe Sandro R. Valentini Rafael V. C. Guido Cleslei F. Zanelli Maysa Furlan
author_facet	Bruna F. Mazzeu Tatiana M. Souza-Moreira Andrew A. Oliveira Melissa Remlinger Lidiane G. Felippe Sandro R. Valentini Rafael V. C. Guido Cleslei F. Zanelli Maysa Furlan
author_sort	Bruna F. Mazzeu
title	The Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity
title_short	The Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity
title_full	The Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity
title_fullStr	The Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity
title_full_unstemmed	The Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity
title_sort	methionine 549 and leucine 552 residues of friedelin synthase from <i>maytenus ilicifolia</i> are important for substrate binding specificity
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/ddc35de5e5134f5c9b696820b9fe8270
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The Methionine 549 and Leucine 552 Residues of Friedelin Synthase from <i>Maytenus ilicifolia</i> Are Important for Substrate Binding Specificity

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