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

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 an EDTA dianhydride (EDTADa)-derived chelator and charged transition ions. Out the five investigated ions - Fe3+, Co2+, Ni2+, Cu2+, Zn2+ best biocatalytic activity PcPAL was achieved when enzyme immobilized Co2+ ion-charged (31.8 ± 1.2 U/g). To explore features this obtained by immobilization, thermostability reusability PAL biocatalyst investigated. maximize surface functionalization aminoalkylated carrier fine-tuned using glycidol as a thinning group beside EDTADa. The maximal yield (YA=103 %) earned EDTADa used in 1 to 24 ratio. reversibility method allowed development regeneration protocol which enables easy reuse functionalized case inactivation.