title: A thermostable feruloyl-esterase from the hemicellulolytic bacterium Thermobacillus xylanilyticus releases phenolic acids from non-pretreated plant cell walls.
abstract: A gene (Tx-est1) encoding a thermostable feruloyl-esterase was isolated from the genome of the gram-positive hemicellulolytic thermophilic bacterium Thermobacillus xylanilyticus. This gene contains an open reading frame of 1,020 bp encoding a protein with molecular mass of 37.4 kDa, similar to feruloyl-esterases from cellulolytic bacteria and fungi. The recombinant enzyme Tx-Est1 was expressed and produced in Escherichia coli. Tx-Est1 contains the conserved putative lipase residues Ser 202, Asp 287, and His 322 which act as catalytic triad in its C-terminus part. Purified Tx-Est1 was active against phenolic acid derivatives and stable at high temperatures. Optimal activity was observed at 65 °C and the optimal pH was around 8.5. The kinetic parameters of the esterase were determined on various substrates. The enzyme displayed activity against methyl esters of hydrocinnamic acids and feruloylated arabino-xylotetraose, exhibiting high specificity and affinity for the latter. Our results showed that Tx-Est1 is a thermostable feruloyl-esterase which could be useful to hydrolyze arabinoxylans from graminaceous plant cell walls as the enzyme is able to release phenolic acids from a lignocellulose biomass.
authors.affiliation: UMR614 Fractionnement des AgroRessources et Environnement, INRA, 51686 Reims, France. harivony.rakotoarivonina@univ-reims.fr
authors.forename: Harivony
authors.initials: H
authors.lastname: Rakotoarivonina
authors.forename: Beatrice
authors.initials: B
authors.lastname: Hermant
authors.forename: Brigitte
authors.initials: B
authors.lastname: Chabbert
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Touzel
authors.forename: Caroline
authors.initials: C
authors.lastname: Remond
mesh_terms: D056449:Bacillales / Q000201:enzymology* / Q000254:growth & development; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism*; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D004950:Esterases / Q000737:chemistry / Q000378:metabolism; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D062385:Hydroxybenzoates / Q000378:metabolism*; D008970:Molecular Weight; D011134:Polysaccharides / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D014908:Triticum / Q000737:chemistry; D014990:Xylans / Q000378:metabolism; D003313:Zea mays / Q000737:chemistry
chemical_list: D062385:Hydroxybenzoates; D011134:Polysaccharides; D011994:Recombinant Proteins; D014990:Xylans; C007916:hemicellulose; C085118:arabinoxylan; D004950:Esterases; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase; C017616:phenolic acid