Exploring the substrate scope of ferulic acid decarboxylase (FDC1) from Saccharomyces cerevisiae

Ferulic acid decarboxylase from Saccharomyces cerevisiae (ScFDC1) was described to possess a novel, prenylated flavin mononucleotide cofactor (prFMN) providing the first enzymatic 1,3-dipolar cycloaddition mechanism. The high tolerance of the enzyme towards several non-natural substrates, combined w...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2019-01, Vol.9 (1), p.647, Article 647
Main Authors: Nagy, Emma Zsófia Aletta, Nagy, Csaba Levente, Filip, Alina, Nagy, Katalin, Gál, Emese, Tőtős, Róbert, Poppe, László, Paizs, Csaba, Bencze, László Csaba
Format: Article
Language:English
Subjects:
Acids
Acrylic acid
Biocatalysts
Cinnamic acid
Computer applications
Decarboxylation
Ferulic acid
Flavin mononucleotide
Phenothiazine
Saccharomyces cerevisiae
Site-directed mutagenesis
Substrate specificity
Yeast
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Ferulic acid decarboxylase from Saccharomyces cerevisiae (ScFDC1) was described to possess a novel, prenylated flavin mononucleotide cofactor (prFMN) providing the first enzymatic 1,3-dipolar cycloaddition mechanism. The high tolerance of the enzyme towards several non-natural substrates, combined with its high quality, atomic resolution structure nominates FDC1 an ideal candidate as flexible biocatalyst for decarboxylation reactions leading to synthetically valuable styrenes. Herein the substrate scope of ScFDC1 is explored on substituted cinnamic acids bearing different functional groups (-OCH , -CF or -Br) at all positions of the phenyl ring (o-, m-, p-) as well as on several biaryl and heteroaryl cinnamic acid analogues or derivatives with extended alkyl chain. It was found that E. coli whole cells expressing recombinant ScFDC1 could transform a large variety of substrates with high conversion, including several bulky aryl and heteroaryl cinnamic acid analogues, that characterize ScFDC1 as versatile and highly efficient biocatalyst. Computational studies revealed energetically favoured inactive binding positions and limited active site accessibility for bulky and non-linear substrates, such as 2-phenylthiazol-4-yl-, phenothiazine-2-yl- and 5-(4-bromophenyl)furan-2-yl) acrylic acids. In accordance with the computational predictions, site-directed mutagenesis of residue I330 provided variants with catalytic activity towards phenothiazine-2-yl acrylic acid and provides a basis for altering the substrate specificity of ScFDC1 by structure based rational design.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-018-36977-x