title: High-level production of recombinant Aspergillus niger cinnamoyl esterase (FAEA) in the methylotrophic yeast Pichia pastoris.
abstract: The cDNA encoding Aspergillus niger cinnamoyl esterase (FAEA) with its native signal sequence was isolated by reverse transcriptase-polymerase chain reaction, sequenced, and expressed in Pichia pastoris. Secretion yields up to 300 mg l(-1) were obtained in buffered medium. The recombinant FAEA was purified to homogeneity using a one-step purification protocol and found to be identical to the native enzyme with respect to size, pI, immunoreactivity and N-terminal sequence. Specific activity, pH and temperature optimum, and kinetic parameters were also found similar to the native esterase. FAEA is thus the first fungal esterase efficiently produced using a heterologous system.
authors.affiliation: Nutrition, Health and Consumer Science Division, Institute of Food Research, Norwich Research Park, Colney, UK.
authors.forename: N
authors.initials: N
authors.lastname: Juge
authors.forename: G
authors.initials: G
authors.lastname: Williamson
authors.forename: A
authors.initials: A
authors.lastname: Puigserver
authors.forename: N J
authors.initials: NJ
authors.lastname: Cummings
authors.forename: I F
authors.initials: IF
authors.lastname: Connerton
authors.forename: C B
authors.initials: CB
authors.lastname: Faulds
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D001709:Biotechnology / Q000379:methods; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D003373:Coumaric Acids / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D007700:Kinetics; D010843:Pichia / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D012331:RNA, Fungal / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D003373:Coumaric Acids; D018076:DNA, Complementary; D012331:RNA, Fungal; D011994:Recombinant Proteins; C004999:ferulic acid; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase