Applied Microbial and Cell Physiology

Applied Microbiology and Biotechnology

, Volume 89, Issue 2, pp 375-385

First online:

Heterologous expression of a thermophilic esterase in Kluyveromyces yeasts

  • Saul Nitsche RochaAffiliated withDepartment of Chemical Engineering, University of São PauloUniversidade Positivo, Rua Pedro Viriato Parigot de Souza
  • , José Abrahão-NetoAffiliated withSchool of Pharmaceutical Sciences, University of São Paulo
  • , María Esperanza CerdánAffiliated withDepartamento de Bioloxía Celular e Molecular, Facultade de Ciencias, Universidade da Coruña
  • , Andreas Karoly GombertAffiliated withDepartment of Chemical Engineering, University of São Paulo
  • , María Isabel González-SisoAffiliated withDepartamento de Bioloxía Celular e Molecular, Facultade de Ciencias, Universidade da Coruña Email author 

Rent the article at a discount

Rent now

* Final gross prices may vary according to local VAT.

Get Access

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 esterase Kluyveromyces marxianus Kluyveromyces lactis Heterologous expression