Immobilization of fuculose-1-phosphate aldolase from Escherichia coli to glyoxal-agarose gels by multipoint covalent attachment

Trinitat Suau, Jordi Calveras, Pere Clapés, Maria Dolors Benaiges, Gregorio Álvaro

    Research output: Contribution to journalArticleResearchpeer-review

    6 Citations (Scopus)

    Abstract

    Recombinant fuculose 1-phosphate aldolase (FucA) from Escherichia coli has been immobilized by multipoint covalent attachment to glyoxal-agarose gels. Experiments, varying the main parameters that control the immobilization process (surface density of aldehyde groups, temperature, pH), were carried out. An immobilization yield of 80-90% and FuCA retained activity on immobilized derivative of 10-20% can be achieved when pH 10, 20°C and 200 μmoleS cm-3 of aldehyde groups was used. The observed activity loss in the immobilization process might be related to the fact that the complex quaternary structure of the enzyme could not be maintained. A short contact-time enzyme support is required to obtain high ratio of active to total immobilized enzyme. A highly loaded derivative of immobilized FucA (65 AU cm-3 of support) has been prepared to use in aldol condensation reactions. Reactions catalyzed by these aldolases involve the use of non-conventional media because of substrate solubility. For instance, the condensation of dihydroxyacetone phosphate (DHAP) and Z-amino-propanal, Z-(R)-alaninal and Z-(S)-alaninal in highly concentrated water-in-oil emulsions gave synthetic yields of 40, 25 and 29% respectively. © 2005 Taylor & Francis.
    Original languageEnglish
    Pages (from-to)241-250
    JournalBiocatalysis and Biotransformation
    Volume23
    DOIs
    Publication statusPublished - 1 May 2005

    Keywords

    • Agarose gel
    • Aldol condensation
    • Covalent attachment
    • Enzyme immobilization
    • Fuculose-1-phosphate aldolase
    • Non-conventional media

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