Open AccessImmobilization of Phenylalanine Ammonia-lyase via EDTA Based Metal Chelate Complexes – Optimization and Prospects
Author(s) -
Evelin SántaBell,
Norbert Krisztián Kovács,
Bálint Alács,
Zsófia Molnár,
Gábor Hornyánszky
Publication year - 2021
Publication title -
periodica polytechnica. chemical engineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.322
H-Index - 19
eISSN - 1587-3765
pISSN - 0324-5853
DOI - 10.3311/ppch.17891
Subject(s) - glycidol , chemistry , chelation , biocatalysis , metal ions in aqueous solution , thermostability , immobilized enzyme , phenylalanine ammonia lyase , surface modification , metal , yield (engineering) , phenylalanine , combinatorial chemistry , chromatography , nuclear chemistry , organic chemistry , enzyme , catalysis , amino acid , biochemistry , materials science , reaction mechanism , peroxidase , metallurgy
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.