The feruloyl esterase system of Talaromyces stipitatus: production of three discrete feruloyl esterases, including a novel enzyme, TsFaeC, with a broad substrate specificity.
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 M(R) 55 340Da. There was no evidence for a carbohydrate-binding domain in TsFaeC.
Nutrition Division, Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK.
Maria-Teresa
MT
Garcia-Conesa
Valérie F
VF
Crepin
Andrew J
AJ
Goldson
Gary
G
Williamson
Nicola J
NJ
Cummings
Ian F
IF
Connerton
Craig B
CB
Faulds
Paul A
PA
Kroon
D000595:Amino Acid Sequence; D002265:Carboxylic Ester Hydrolases / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry* / Q000145:classification; D004789:Enzyme Activation; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013379:Substrate Specificity; D032901:Talaromyces / Q000201:enzymology*; D013696:Temperature
D007527:Isoenzymes; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase