The feruloyl esterase system of Talaromyces stipitatus: Production of three discrete feruloyl esterases, including a novel enzyme, TsFaeC, with a broad substrate specificity

Maria Teresa Garcia-Conesa, Valérie F. Crepin, Andrew J. Goldson, Gary Williamson, Nicola J. Cummings, Ian F. Connerton, Craig B. Faulds, Paul A. Kroon

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Abstract

Several extracellular feruloyl esterases were produced by the mesophilic fungus Talaromyces stipitatus when grown on selective carbon sources in liquid media. Type-A and Type-B feruloyl esterases, as defined by their substrate specificity against methyl hydroxycinnamates, were produced during growth on wheat bran and sugar beet pulp, respectively. In addition, Tal. stipitatus produced a new type of esterase (TsFaeC) during growth on sugar beet pulp with a broader spectrum of activity (Type-C) against the (hydroxy)cinnamate esters than those previously described. All three enzymes were purified and N-terminal amino acid sequences and internal peptide sequences determined. The TsFaeC sequences were used to amplify a gene fragment from Tal. stipitatus genomic DNA. The flanking sequences were identified with the aid of RACE-RTPCR, and a full-length clone constructed. The faeC gene is present as a single copy and contains a single intron. The complete cDNA fragment contains an ORF of 1590bp, faeC, which is predicted to encode a 530 amino acid pre-protein, including a 25-residue signal peptide, and to produce a mature protein of MR 55 340Da. There was no evidence for a carbohydrate-binding domain in TsFaeC.

Original languageEnglish
Pages (from-to)227-241
Number of pages15
JournalJournal of Biotechnology
Volume108
Issue number3
DOIs
Publication statusPublished - 18 Mar 2004
Externally publishedYes

Keywords

  • [2-O-(trans-feruloyl)-α-L-arabinofuranosyl]-(1,5)-L- arabinofuranose
  • [5-O-(trans-feruloyl)-α-L-arabinofuranosyl]-(1,3)- β-D-xylopyranosyl-(1,4)-D-xylopyranose
  • AraF
  • FAXX
  • MCA
  • Methyl caffeate (methyl 3,4-dihydroxycinnamate)

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