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Applied Microbiology and Biotechnology

, Volume 79, Issue 6, pp 1001–1008 | Cite as

High-level overproduction of Thermus enzymes in Streptomyces lividans

  • Margarita Díaz
  • Eloy Ferreras
  • Renata Moreno
  • Ana Yepes
  • José Berenguer
  • Ramón Santamaría
Applied Genetics and Molecular Biotechnology

Abstract

Biotechnology needs to explore the capacity of different organisms to overproduce proteins of interest at low cost. In this paper, we show that Streptomyces lividans is a suitable host for the expression of Thermus thermophilus genes and report the overproduction of the corresponding proteins. This capacity was corroborated after cloning the genes corresponding to an alkaline phosphatase (a periplasmic enzyme in T. thermophilus) and that corresponding to a beta-glycosidase (an intracellular enzyme) in Escherichia coli and in S. lividans. Comparison of the production in both hosts revealed that the expression of active protein achieved in S. lividans was much higher than in E. coli, especially in the case of the periplasmic enzyme. In fact, the native signal peptide of the T. thermophilus phosphatase was functional in S. lividans, being processed at the same peptide bond in both organisms, allowing the overproduction and secretion of this protein to the S. lividans culture supernatant. As in E. coli, the thermostability of the expressed proteins allowed a huge purification factor upon thermal denaturation and precipitation of the host proteins. We conclude that S. lividans is a very efficient and industry-friendly host for the expression of thermophilic proteins from Thermus spp.

Keywords

Xylose Streptomyces Codon Usage Coomassie Blue Staining Thermostable Enzyme 
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.

Notes

Acknowledgments

This work has been supported by grants CSI02A05 from the Junta de Castilla y León to R. Santamaría and BIO2007-60245 and S0505/PPQ/0344 from the Ministry of Education and Science and the Comunidad Autónoma de Madrid, respectively, to J. Berenguer. Institutional grants from Fundación Ramón Areces to CBMSO and from Junta de Castilla y León are also acknowledged. We thank MJ Jimenez Rufo for her excellent technical work. Thanks are also due to N. Skinner for supervising the English version of the manuscript.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Margarita Díaz
    • 1
    • 3
  • Eloy Ferreras
    • 2
  • Renata Moreno
    • 2
  • Ana Yepes
    • 1
  • José Berenguer
    • 2
  • Ramón Santamaría
    • 1
  1. 1.Instituto de Microbiología Bioquímica, Departamento de Microbiología y Genética, Consejo Superior de Investigaciones Científicas (CSIC)Universidad de SalamancaSalamancaSpain
  2. 2.Departamento de Biología Molecular, Centro de Biología Molecular Severo OchoaUniversidad Autónoma de MadridMadridSpain
  3. 3.Instituto de Microbiología Bioquímica, CSIC /Universidad de Salamanca, Edificio DepartamentalSalamancaSpain

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