Redesign of the Phosphate Binding Site of L-Rhamnulose- 1-Phosphate Aldolase towards a Dihydroxyacetone Dependent Aldolase

Authors

  • Xavier Garrabou,

    1. Department of Biological Chemistry and Molecular Modeling, Instituto de Química Avanzada de Cataluña – CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain, Fax: (+34)-93-204-5904; phone: (+34)-93-400-6112
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  • Jesús Joglar,

    1. Department of Biological Chemistry and Molecular Modeling, Instituto de Química Avanzada de Cataluña – CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain, Fax: (+34)-93-204-5904; phone: (+34)-93-400-6112
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  • Teodor Parella,

    1. Servei de Ressonància Magnètica Nuclear, Department of Chemistry, Universitat Autònoma de Barcelona, Bellaterra, Spain
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  • Ramon Crehuet,

    1. Department of Biological Chemistry and Molecular Modeling, Instituto de Química Avanzada de Cataluña – CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain, Fax: (+34)-93-204-5904; phone: (+34)-93-400-6112
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  • Jordi Bujons,

    1. Department of Biological Chemistry and Molecular Modeling, Instituto de Química Avanzada de Cataluña – CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain, Fax: (+34)-93-204-5904; phone: (+34)-93-400-6112
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  • Pere Clapés

    Corresponding author
    1. Department of Biological Chemistry and Molecular Modeling, Instituto de Química Avanzada de Cataluña – CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain, Fax: (+34)-93-204-5904; phone: (+34)-93-400-6112
    • Department of Biological Chemistry and Molecular Modeling, Instituto de Química Avanzada de Cataluña – CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain, Fax: (+34)-93-204-5904; phone: (+34)-93-400-6112
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Abstract

The aldol addition of unphosphorylated dihydroxyacetone (DHA) to aldehydes catalyzed by L-rhamnulose-1-phosphate aldolase (RhuA), a dihydroxyacetone phosphate-dependent aldolase, is reported. Moreover, a single point mutation in the phosphate binding site of the RhuA wild type, that is, substitution of aspartate for asparagine at position N29, increased by 3-fold the equation image of aldol addition reactions of DHA to other aldehyde acceptors rather than the natural L-lactaldehyde. The RhuA N29D mutant modified the optimum enzyme design for the natural substrate and changed its catalytic properties making the aldolase more versatile to other aldol additions of DHA.

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