Applied Microbiology and Biotechnology

, Volume 87, Issue 5, pp 1753–1764 | Cite as

Hydrolysis of cyclic poly(ethylene terephthalate) trimers by a carboxylesterase from Thermobifida fusca KW3

  • Susan Billig
  • Thorsten Oeser
  • Claudia Birkemeyer
  • Wolfgang Zimmermann
Biotechnologically Relevant Enzymes and Proteins


We have identified a carboxylesterase produced in liquid cultures of the thermophilic actinomycete Thermobifida fusca KW3 that were supplemented with poly(ethylene terephthalate) fibers. The enzyme hydrolyzed highly hydrophobic, synthetic cyclic poly(ethylene terephthalate) trimers with an optimal activity at 60°C and a pH of 6. V max and K m values for the hydrolysis were 9.3 µmol−1 min−1 mg−1 and 0.5 mM, respectively. The esterase showed high specificity towards short and middle chain-length fatty acyl esters of p-nitrophenol. The enzyme retained 37% of its activity after 96 h of incubation at 50°C and a pH of 8. Enzyme inhibition studies and analysis of substitution mutants of the carboxylesterase revealed the typical catalytic mechanism of a serine hydrolase with a catalytic triad composed of serine, glutamic acid, and histidine.


Thermobifida fusca Carboxylesterase Cutinase Poly(ethylene terephthalate) (PET) Enzymatic hydrolysis 



S. Billig was supported by grant no. 20004/730, Deutsche Bundesstiftung Umwelt and T. Oeser by grant no. 13-8811.61/215-1, Sächsisches Staatsministerium für Umwelt und Landwirtschaft. C. Roth from the University of Leipzig is acknowledged for his assistance in the pI determination. I. Neundorf and J. Stichel from the University of Leipzig are acknowledged for the amino acid sequence determination and MALDI-TOF/TOF analysis.

Supplementary material

253_2010_2635_MOESM1_ESM.pdf (6 kb)
Online resource 1 Amino acid composition of TfCa and the cutinases Tfu_0882 and Tfu_0883 from T. fusca YX (PDF 6 kb)
253_2010_2635_MOESM2_ESM.pdf (63 kb)
Online resource 2 Comparison of the amino acid sequence of TfCa from T. fusca KW3 with the putative carboxylesterase Tfu_2427 from T. fusca YX. The proteins differ in two amino acids (marked in grey and bold). Tfu_2427 has a glycine residue instead of a serine at position 335 and a threonine residue instead of an alanine at position 340. The identity of the two proteins was 99.6%. Amino acid residues of the catalytic triad are underlined and marked in bold (PDF 63 kb)
253_2010_2635_MOESM3_ESM.pdf (1.7 mb)
Online resource 3 Alignment of amino acid sequences of carboxylesterases (PDF 1748 kb)


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Susan Billig
    • 1
  • Thorsten Oeser
    • 1
  • Claudia Birkemeyer
    • 2
  • Wolfgang Zimmermann
    • 1
  1. 1.Department of Microbiology and Bioprocess Technology, Institute of BiochemistryUniversity of LeipzigLeipzigGermany
  2. 2.Institute of Analytical ChemistryUniversity of LeipzigLeipzigGermany

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