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High-level overproduction of Thermus enzymes in Streptomyces lividans

  • Applied Genetics and Molecular Biotechnology
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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.

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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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Authors and Affiliations

  1. Instituto de Microbiología Bioquímica, Departamento de Microbiología y Genética, Consejo Superior de Investigaciones Científicas (CSIC), Universidad de Salamanca, Edificio Departamental, Campus Miguel de Unamuno, 37007, Salamanca, Spain

    Margarita Díaz, Ana Yepes & Ramón Santamaría

  2. Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa, Universidad Autónoma de Madrid, 28049, Madrid, Spain

    Eloy Ferreras, Renata Moreno & José Berenguer

  3. Instituto de Microbiología Bioquímica, CSIC /Universidad de Salamanca, Edificio Departamental, Campus Miguel de Unamuno, 37007, Salamanca, Spain

    Margarita Díaz

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  1. Margarita Díaz
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  2. Eloy Ferreras
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  3. Renata Moreno
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  4. Ana Yepes
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  5. José Berenguer
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  6. Ramón Santamaría
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Correspondence to Margarita Díaz.

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Díaz, M., Ferreras, E., Moreno, R. et al. High-level overproduction of Thermus enzymes in Streptomyces lividans . Appl Microbiol Biotechnol 79, 1001–1008 (2008). https://doi.org/10.1007/s00253-008-1495-1

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  • DOI: https://doi.org/10.1007/s00253-008-1495-1

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