Immobilization of Phenylalanine Ammonia-lyase via EDTA Based Metal Chelate Complexes – Optimization and Prospects

Authors

  • Evelin Sánta-Bell
    Affiliation

    Department of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, H-1111 Budapest, Műegyetem rkp. 3., Hungary

  • Norbert Krisztián Kovács
    Affiliation

    Department of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, H-1111 Budapest, Műegyetem rkp. 3., Hungary;

    Fermentia Ltd, H-1045 Budapest, Berlini u. 47–49., Hungary

  • Bálint Alács
    Affiliation

    Department of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, H-1111 Budapest, Műegyetem rkp. 3., Hungary

  • Zsófia Molnár
    Affiliation

    Department of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, H-1111 Budapest, Műegyetem rkp. 3., Hungary;

    Institute of Enzymology, ELKH-Research Center for Natural Sciences, H-1117 Budapest, Magyar tudósok krt. 2., Hungary

  • Gábor Hornyánszky
    Affiliation

    Department of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, H-1111 Budapest, Műegyetem rkp. 3., Hungary;

    SynBiocat Ltd., H-1172 Budapest, Szilasliget u. 3., Hungary

https://doi.org/10.3311/PPch.17891

Abstract

Immobilized metal ion affinity chromatography principles were applied for selective immobilization of recombinant polyhistidine tag fused phenylalanine ammonia-lyase from parsley (PcPAL) on porous polymeric support with aminoalkyl moieties modified with an EDTA dianhydride (EDTADa)-derived chelator and charged with transition metal ions. Out of the five investigated metal ions - Fe3+, Co2+, Ni2+, Cu2+, Zn2+ - the best biocatalytic activity of PcPAL was achieved when the enzyme was immobilized on the Co2+ ion-charged support (31.8 ± 1.2 U/g). To explore the features this PcPAL obtained by selective immobilization, the thermostability and reusability of this PAL biocatalyst were investigated. To maximize the activity of the immobilized PcPAL the surface functionalization of the aminoalkylated polymeric carrier was fine-tuned with using glycidol as a thinning group beside EDTADa. The maximal activity yield (YA=103 %) was earned when the EDTADa and glycidol were used in 1 to 24 ratio. The reversibility of the immobilization method allowed the development of a support regeneration protocol which enables easy reuse of the functionalized support in case of enzyme inactivation.

Keywords:

IMAC, selective enzyme immobilization, phenylalanine ammonia-lyase

Published Online

2021-05-27

How to Cite

Sánta-Bell, E., Kovács, N. K., Alács, B., Molnár, Z., Hornyánszky, G. “Immobilization of Phenylalanine Ammonia-lyase via EDTA Based Metal Chelate Complexes – Optimization and Prospects”, Periodica Polytechnica Chemical Engineering, 65(3), pp. 308–319, 2021. https://doi.org/10.3311/PPch.17891

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Articles