Immobilization of phenylalanine ammonia-lyase on hydroxyapatite and hydroxyapatite composites (original) (raw)
Related papers
Molecules, 2019
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.
Periodica Polytechnica Chemical Engineering, 2017
A new and efficient immobilized form of phenylalanine ammonia-lyase (PAL) was obtained by covalent linkage onto amino functionalized single-walled carbon nanotubes (SwCNTNH2) as carrier. The catalytic properties of the resulted nanostructured biocatalyst (SwCNTNH2-PAL) were tested in the kinetic resolution of racemic 2-amino-3-(thiophen-2-yl)propanoic acid 1 by ammonia elimination and in the enantiotope selective addition of ammonia onto (E)-3-(thiophen-2-yl)acrylic acid 2. SwCNTNH2-PAL was a durable biocatalyst in batch mode for ammonia elimination from 1 (>85% of original activity after 7 cycles) and in ammonia addition to 2 (>70% of original activity after 3 cycles in 6 M NH3 , pH 10.0). The ammonia addition onto 2 was also studied in a continuous-flow microreactor packed with SwCNTNH2-PAL (2 M NH3, pH 10.0, 15 bar) in the 30-80 °C temperature range. No significant loss of PAL activity was observed over 72 h in the microreactor up to 60 °C. Productivity of SwCNTNH2-PAL at 3...
Catalysts, 2020
Currently, great attention is focused on conducting manufacture processes using clean and eco-friendly technologies. This research trend also relates to the production of immobilized biocatalysts of industrial importance using matrices and methods that fulfill specified operational and environmental requirements. For that reason, hydrogels of natural origin and the entrapment method become increasingly popular in terms of enzyme immobilization. The presented work is the comparative research on invertase immobilization using two natural hydrogel matrices—alginate and gelatin. During the study, we provided the molecular insight into the structural characteristics of both materials regarding their applicability as effective enzyme carriers. In order to confirm our predictions of using these hydrogels for invertase immobilization, we performed the typical experimental studies. In this case, the appropriate conditions of enzyme entrapment were selected for both types of carrier. Next, th...
Enzyme immobilization onto renewable polymeric matrixes: Past, present, and future trends
Journal of Applied Polymer Science, 2015
ABSTRACTIn this review, we present an overview of the different renewable polymers that are currently being used as matrixes for enzyme immobilization and their properties and of new developments in biocatalysts preparation and applications. Polymers obtained from renewable resources have attracted much attention in recent years because they are environmentally friendly and available in large quantities from natural sources. Different methods for the immobilization of enzymes with these matrixes are reviewed, in particular: (1) binding to a prefabricated biopolymer, (2) entrapment, and (3) crosslinking of enzyme molecules. Emphasis is given to relatively recent developments, such as the use of novel supports, novel entrapment methods and protocols of polymer derivatization, and the crosslinking of enzymes. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42125.
Food chemistry, 2018
Stability of enzymes is an important parameter for their industrial applicability. Here, we report successful immobilization of β-amylase (bamyl) from peanut (Arachis hypogaea) onto Graphene oxide-carbon nanotube composite (GO-CNT), Graphene oxide nanosheets (GO) and Iron oxide nanoparticles (FeO). The Box-Behnken Design of Response Surface Methodology (RSM) was used which optimized parameters affecting immobilization and gave 90%, 88% and 71% immobilization efficiency, respectively, for the above matrices. β-Amylase immobilization onto GO-CNT (bamyl@GO-CNT) and FeO(bamyl@FeO), resulted into approximately 70% retention of activity at 65 °C after 100 min of exposure. We used atomic force microscopy (AFM), scanning and transmission electron microscopy (SEM and TEM), Fourier transformed infrared (FT-IR) spectroscopy and fluorescence microscopy for characterization of free and enzyme bound nanostructures (NS). Due to the non-toxic nature of immobilization matrices and simple but elegant...
Modified chitosan microspheres in non-aggregated amylase immobilization
International Journal of Biological Macromolecules, 2014
Immobilized enzymes are useful as reusable catalysts in industrial processes. In this study, ␣-amylase was used as a model enzyme to evaluate the propensity of synthesized porous chitosan microspheres as immobilization matrix. Chitosan microspheres were synthesized by grafting and covalent gelation technique using acrylamide (AAm) and glutaraldehyde (GA) as chemical agents, respectively. The synthesized chitosan-cl-poly(AAm) demonstrated amylase immobilization capacity of 350 mg/g. Furthermore, SEM results supported the porous microsphere structure for chitosan-cl-poly(AAm) with non-aggregated amylase immobilization, which accounts for comparable activity of immobilized amylase (3.28 mol/ml/min) in contrast to free amylase (3.46 mol/ml/min). The immobilized ␣-amylase was characterized for optimal pH and temperature activity and showed better resistance to temperature and pH inactivation in contrast to free amylase. The immobilized amylase retained more than 60% of its initial activity when stored at 4 • C for 30 days and retained 50% of its initial activity after seven successive repeated-use cycles. In conclusion, the study can be used as base for the immobilization of competent industrial biocatalysts in non-aggregated active structure.
Use of Nanomaterials for the Immobilization of Industrially Important Enzymes
Immobilization enables enzymes to be held in place so that they can be easily separated from the product when needed and can be used again. Conventional methods of immobilization include adsorption, encapsulation, entrapment, cross-linking and covalent binding. However, conventional methods have several drawbacks including reduced stability, loss of biomolecules, less enzyme loading or activity and limited diffusion. The aim of this study is the evaluation of importance of nanomaterials for the immobilization of industrially important enzymes. Nano materials are now in trend for the immobilization of different enzymes due to their physi-chemical properties. Gold nanoparticles, silver nanoparticles, nano diamonds, graphene, carbon nanotubes and others are used for immobilization. Among covalent and non-covalent immobilization of enzymes involving both single and multiwalled carbon nanotubes, non-covalent immobilization with functionalized carbon nanotubes is superior. There-fore, enzymes immobilized with nanomaterials possess greater stability, retention of catalytic activi-ty and reusability of enzymes.
Catalysts, 2021
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...