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Applied Microbiology and Biotechnology

, Volume 63, Issue 6, pp 686–690 | Cite as

Purification and characterization of a type B feruloyl esterase (StFAE-A) from the thermophilic fungus Sporotrichum thermophile

  • E. Topakas
  • H. Stamatis
  • P. Biely
  • P. Christakopoulos
Short Contribution

Abstract

A feruloyl esterase (StFAE-A) produced by Sporotrichum thermophile was purified to homogeneity. The purified homogeneous preparation of native StFAE-A exhibited a molecular mass of 57.0±1.5 kDa, with a mass of 33±1 kDa on SDS-PAGE. The pI of the enzyme was estimated by cation-exchange chromatofocusing to be at pH 3.1. The enzyme activity was optimal at pH 6.0 and 55–60 °C. The purified esterase was stable at the pH range 5.0–7.0. The enzyme retained 70% of activity after 7 h at 50 °C and lost 50% of its activity after 45 min at 55 °C and after 12 min at 60 °C. Determination of k cat/K m revealed that the enzyme hydrolyzed methyl p-coumarate 2.5- and 12-fold more efficiently than methyl caffeate and methyl ferulate, respectively. No activity on methyl sinapinate was detected. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose and it hydrolyzed 4-nitrophenyl 5-O-trans-feruloyl-α-l-arabinofuranoside (NPh-5-Fe-Araf) 2-fold more efficiently than NPh-2-Fe-Araf. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with xylanase from S. thermophile (a maximum of 34% total ferulic acid released after 1 h incubation). StFAE-A by itself could release FA, but at a level almost 47-fold lower than that obtained in the presence of xylanase. The potential of StFAE-A for the synthesis of various phenolic acid esters was tested using a ternary water-organic mixture consisting of n-hexane, 1-butanol and water as a reaction system.

Keywords

Ferulic Acid Cinnamic Acid Butyl Ester Thermophilic Fungus Feruloyl Esterase 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • E. Topakas
    • 1
  • H. Stamatis
    • 2
  • P. Biely
    • 3
  • P. Christakopoulos
    • 1
    • 4
  1. 1.Biotechnology Laboratory, Chemical Engineering DepartmentNational Technical University of AthensAthensGreece
  2. 2.Biotechnology Laboratory, Biochemical Applications and Technologies DepartmentUniversity of IoanninaIoanninaGreece
  3. 3.Institute of ChemistrySlovak Academy of SciencesBratislavaSlovak Republic
  4. 4.Center for Process BiotechnologyTechnical University of DenmarkLyngbyDenmark

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