Entrapment of Phenylalanine Ammonia-Lyase in Nanofibrous Polylactic Acid Matrices by Emulsion Electrospinning

Gábor Koplányi,Ferenc Ender,Muriel Józó,Zsófia Molnár,Beáta G Vértessy,Béla Pukánszky,László Poppe,Gergő Dániel Tóth,Diána Balogh-Weiser,András Szilágyi,Evelin Sánta-Bell

doi:10.3390/catal11101149

Abstract

Immobilization of the recombinant, plant-derived Petroselinum crispum phenylalanine ammonia lyase (PcPAL) in electrospun matrices have the potential to create promising, easy-to-use biocatalysts. Polylactic acid (PLA) a biologically inert, commercial biopolymer, was chosen as the material of the carrier system. PLA could be electrospun properly only from water-immiscible organic solvents, which limits its application as a carrier of sensitive biological objects. The emulsion electrospinning is a proper solution to overcome this issue using non-ionic emulsifiers with different hydrophilic-lipophilic balance (HLB) values. The stabilized emulsion could protect the sensitive PcPAL dissolved in the aqueous buffer phase and improve fiber formation, plus help to keep the biocatalytic activity of enzymes. In this study, the first approach is described to produce PLA nanofibers containing PcPAL enzymes by emulsion electrospinning and to use the resulted biocatalyst in the ammonia elimination reaction from l-phenylalanine.

Highlights

Enantiopure amino acids have great importance in every aspect of our lives, especially as dietary supplements, essential chiral building elements of peptides, or pharmaceuticals, amino acid-related biotransformation has increasing significance [1,2]
To compare the precursor systems with various Petroselinum crispum phenylalanine ammonia lyase (PcPAL) contents, the viscosity of precursors was determined by rotational rheometer, and the morphology of the final fibrous materials was characterized by scanning electron microscope (SEM)
6:1, v/v) and the aqueous solution of PcPAL are immiscible phases, which can be applied for emulsion electrospinning

Summary

Introduction

Enantiopure amino acids have great importance in every aspect of our lives, especially as dietary supplements, essential chiral building elements of peptides, or pharmaceuticals, amino acid-related biotransformation has increasing significance [1,2]. An industrial scale enzyme is commonly synthesized by a variety of host cells, often by the Escherichia coli (E. coli) bacterium, isolated from the highly complex cellular environment [9,10]. Catalysts 2021, 11, 1149 coli) bacterium, isolated from the highly complex cellular environment [9,10] These so-called processes aretechnological expensive andchallenges have technological in many processesdownstream are expensive and have in many challenges aspects, including aspects, including the formation of inclusion bodies, the structural sensitivity of proteins, the formation of inclusion bodies, the structural sensitivity of proteins, and long-term and long-term storage [11,12,13].storage [11,12,13]

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Conclusion