“Fishing and Hunting”—Selective Immobilization of a Recombinant Phenylalanine Ammonia-Lyase from Fermentation Media

• Main Authors: Evelin Sánta-Bell, Zsófia Molnár, Andrea Varga, Flóra Nagy, Gábor Hornyánszky, Csaba Paizs, Diána Balogh-Weiser, László Poppe
• Format: Article
• Language: English
• Published: MDPI AG 2019-11-01
• Series: Molecules
• Subjects: imac; selective enzyme immobilization; phenylalanine ammonia-lyase; magnetic nanoparticles
• Online Access: https://www.mdpi.com/1420-3049/24/22/4146
• ISSN: 1420-3049

This article overviews the numerous immobilization methods available for various biocatalysts such as whole-cells, cell fragments, lysates or enzymes which do not require preliminary enzyme purification and introduces an advanced approach avoiding the costly and time consuming downstream processes required by immobilization of purified enzyme-based biocatalysts (such as enzyme purification by chromatographic methods and dialysis). Our approach is based on silica shell coated magnetic nanoparticles as solid carriers decorated with mixed functions having either coordinative binding ability (a metal ion complexed by a chelator anchored to the surface) or covalent bond-forming ability (an epoxide attached to the surface via a proper linker) enabling a single operation enrichment and immobilization of a recombinant phenylalanine ammonia-lyase from parsley fused to a polyhistidine affinity tag.