Applied Microbiology and Biotechnology

, Volume 89, Issue 2, pp 375–385

Heterologous expression of a thermophilic esterase in Kluyveromyces yeasts

  • Saul Nitsche Rocha
  • José Abrahão-Neto
  • María Esperanza Cerdán
  • Andreas Karoly Gombert
  • María Isabel González-Siso
Applied Microbial and Cell Physiology

DOI: 10.1007/s00253-010-2869-8

Cite this article as:
Rocha, S.N., Abrahão-Neto, J., Cerdán, M.E. et al. Appl Microbiol Biotechnol (2011) 89: 375. doi:10.1007/s00253-010-2869-8

Abstract

In the present work, a thermophilic esterase from Thermus thermophilus HB27 was cloned into Kluyveromyces marxianus and into Kluyveromyces lactis using two different expression systems, yielding four recombinant strains. K. lactis showed the highest esterase expression levels (294 units per gram dry cell weight, with 65% of cell-bound enzyme) using an episomal system with the PGK promoter and terminator from Saccharomyces cerevisiae combined with the K. lactis k1 secretion signal. K. marxianus showed higher secretion efficiency of the heterologous esterase (56.9 units per gram dry cell weight, with 34% of cell-bound enzyme) than K. lactis. Hydrolytic activities for the heterologous esterases were maximum at pH values between 8.0 and 9.0 for both yeast species and at temperatures of 50 °C and 45 °C for K. marxianus and K. lactis, respectively. When compared to previously published data on this same esterase produced in the original host or in S. cerevisiae, our results indicate that Kluyveromyces yeasts can be considered good hosts for the heterologous secretion of thermophilic esterases, which have a potential application in biodiesel production or in resolving racemates.

Keywords

Thermophilic esteraseKluyveromyces marxianusKluyveromyces lactisHeterologous expression

Supplementary material

253_2010_2869_MOESM1_ESM.pdf (27 kb)
ESM 1(PDF 27 kb)

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Saul Nitsche Rocha
    • 1
    • 4
  • José Abrahão-Neto
    • 2
  • María Esperanza Cerdán
    • 3
  • Andreas Karoly Gombert
    • 1
  • María Isabel González-Siso
    • 3
  1. 1.Department of Chemical EngineeringUniversity of São PauloSão PauloBrazil
  2. 2.School of Pharmaceutical SciencesUniversity of São PauloSão PauloBrazil
  3. 3.Departamento de Bioloxía Celular e Molecular, Facultade de CienciasUniversidade da CoruñaCoruñaSpain
  4. 4.Universidade PositivoRua Pedro Viriato Parigot de SouzaCuritibaBrasil