High-level production of recombinant Aspergillus niger cinnamoyl esterase (FAEA) in the methylotrophic yeast Pichia pastoris

Nathalie Juge, Gary Williamson, Antoine Puigserver, Nicola J. Cummings, Ian F. Connerton, Craig B. Faulds

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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.

Original languageEnglish
Pages (from-to)127-132
Number of pages6
JournalFEMS Yeast Research
Volume1
Issue number2
DOIs
Publication statusPublished - 1 Dec 2001
Externally publishedYes

Keywords

  • Aspergillus niger
  • Cinnamoyl esterase
  • Ferulic acid
  • Heterologous expression
  • Pichia pastoris
  • Secretion

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