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Structural and functional analysis of hybrid enzymes generated by domain shuffling between Saccharomyces cerevisiae (var. diastaticus) Sta1 glucoamylase and Saccharomycopsis fibuligera Bgl1 β-glucosidase

  • Applied Genetics and Molecular Biotechnology
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Abstract

Saccharomyces cerevisiae Sta1 glucoamylase and Saccharomycopsis fibuligera Bgl1 β-glucosidase, two relevant enzymes from a biotechnological point of view, are proteins with multidomain structure. Starting with homology-based structural models of Sta1 and Bgl1, we have constructed a series of hybrid enzymes by interchanging domains of the two proteins. The first purpose of these constructs was to check available hypotheses about the uncertain biological functions of two domains: the serine/threonine-rich domain (STRD) of Sta1 and a β-sandwich domain present in Bgl1 that we have designated fibronectin-like domain (FLD). While, according to the initial hypothesis, proteins carrying the FLD tend to adhere to the cell wall, our results argued against the idea of an involvement of the STRD in protein secretion that stemmed from the presence of similar domains in different proteins secreted by yeast. The second objective of this work was to increase the enzymatic repertoire by generating enzymes with new structural and functional properties.

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Acknowledgements

This work was funded by Spanish Ministerio de Ciencia e Innnovación grant BIO2007-67708-C04-02. Leontina Gurgu was supported by a short term Collaborative Experimental Scholarship for Central & Eastern Europe from Federation of European Biochemical Societies (FEBS) and by Romanian project PNCDI II IDEI COD CNCSIS 658 No. 315/1.10.2007 awarded to Dr. Vasilica Barbu.

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Authors and Affiliations

  1. Instituto de Agroquímica y Tecnología de Alimentos, CSIC, Paterna, Valencia, Spain

    Julia Marín-Navarro, Leontina Gurgu, Santiago Alamar & Julio Polaina

  2. Faculty of Food Science and Engineering, Dunarea de Jos University, Galati, Romania

    Leontina Gurgu

  3. Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Científicas, Av. Agustín Escardino 7, E46980, Paterna, Valencia, Spain

    Julio Polaina

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  1. Julia Marín-Navarro
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  2. Leontina Gurgu
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  3. Santiago Alamar
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  4. Julio Polaina
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Correspondence to Julio Polaina.

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Marín-Navarro, J., Gurgu, L., Alamar, S. et al. Structural and functional analysis of hybrid enzymes generated by domain shuffling between Saccharomyces cerevisiae (var. diastaticus) Sta1 glucoamylase and Saccharomycopsis fibuligera Bgl1 β-glucosidase. Appl Microbiol Biotechnol 89, 121–130 (2011). https://doi.org/10.1007/s00253-010-2845-3

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