TY - JOUR AU - Alács, Bálint AU - Zrinyi, Anna AU - Hornyánszky, Gábor AU - Poppe, László AU - Bell, Evelin TI - Upgrading Epoxy Supports for Enzyme Immobilization by Affinity Function Doping—A Case Study with Phenylalanine Ammonia-Lyase from Petroselinum crispum JF - CATALYSTS J2 - CATALYSTS VL - 14 PY - 2024 IS - 1 PG - 15 SN - 2073-4344 DO - 10.3390/catal14010014 UR - https://m2.mtmt.hu/api/publication/34451217 ID - 34451217 N1 - Department of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest, H-1111, Hungary SynBiocat Ltd., Szilasliget u 3, Budapest, H-1172, Hungary Biocatalysis and Biotransformation Research Centre, Faculty of Chemistry and Chemical Engineering, Babes-Bolyai University of Cluj-Napoca, Arany János Str. 11, Cluj-Napoca, RO-400028, Romania Export Date: 9 February 2024 Correspondence Address: Poppe, L.; Department of Organic Chemistry and Technology, Műegyetem rkp. 3, Hungary; email: poppe.laszlo@vbk.bme.hu Correspondence Address: Bell, E.; Department of Organic Chemistry and Technology, Műegyetem rkp. 3, Hungary; email: bell.evelin@vbk.bme.hu Funding details: European Commission, EC Funding details: Richter Gedeon Talentum Alapítvány Funding details: Nemzeti Kutatási, Fejlesztési és Innovaciós Alap, NKFIA, RRF-2.3.1-21-2022-00015 Funding details: Innovációs és Technológiai Minisztérium Funding text 1: B.A. acknowledges the support of Gedeon Richter Talentum Foundation and E.B., for the support of the Mihály Somogyi grant of the Jozsef Varga Foundation. Funding text 2: The research reported in this paper is part of project no. TKP2021-EGA-02, implemented with the support provided by the Ministry for Innovation and Technology of Hungary from the National Research, Development, and Innovation Fund, financed under the TKP2021 funding scheme. The research was performed in the frame of project no. RRF-2.3.1-21-2022-00015 and has been implemented with the support provided by the European Union. AB - This article provides a method to upgrade epoxy-functionalized carriers for covalent enzyme immobilization to selective carriers suitable for covalent immobilization of metal affinity-tagged enzymes without the need of preliminary enzyme purification. Affinity function doping of the epoxy-functionalized surface introduces an advanced possibility to avoid the costly and time-consuming downstream processes required for efficient immobilization on non-selective epoxy carriers. Our approach is based on the partial functionalization of surface epoxides via a proper diamine-derived linker and an ethylenediaminetetraacetic dianhydride-based chelator charged with cobalt ions. The solid macroporous carriers, doped with metal affinity functions, have both coordinative binding ability (rapid anchoring the metal affinity-tagged enzymes to the surface) and subsequent covalent bond-forming ability (preferred binding of the tagged enzyme to the surface after proper washing by the residual epoxide functions), enabling a single operation for the enrichment and immobilization of a recombinant phenylalanine ammonia-lyase from parsley fused to a polyhistidine affinity tag. The immobilized PcPAL was applied in the ammonia elimination of racemic phenylalanine, 4-chlorophenylalanine, and 4-bromophenylalanine to produce the corresponding d-phenylalanines, in addition to the formation of (E)-cinnamates, as well as in ammonia addition reactions to (E)-cinnamates, yielding the corresponding enantiopure l-phenylalanines. LA - English DB - MTMT ER - TY - JOUR AU - Koplányi, Gábor AU - Bell, Evelin AU - Molnár, Zsófia Klára AU - Katona, Gábor AU - Neumann, Péter Lajos AU - Ender, Ferenc AU - Balogh, György Tibor AU - Žnidaršič-Plazl, Polona AU - Poppe, László AU - Balogh Weiser, Diána TI - Novel Approach for the Isolation and Immobilization of a Recombinant Transaminase. Applying an Advanced Nanocomposite System TS - Applying an Advanced Nanocomposite System JF - CHEMBIOCHEM J2 - CHEMBIOCHEM VL - 24 PY - 2023 IS - 7 PG - 11 SN - 1439-4227 DO - 10.1002/cbic.202200713 UR - https://m2.mtmt.hu/api/publication/33574611 ID - 33574611 LA - English DB - MTMT ER - TY - JOUR AU - Imarah, Ali O. AU - Silva, Fausto M. W. G. AU - Tuba, László AU - Malta-Lakó, Ágnes AU - Szemes, József AU - Bell, Evelin AU - Poppe, László TI - A Convenient U-Shape Microreactor for Continuous Flow Biocatalysis with Enzyme-Coated Magnetic Nanoparticles–Lipase-Catalyzed Enantiomer Selective Acylation of 4-(Morpholin-4-yl)butan-2-ol JF - CATALYSTS J2 - CATALYSTS VL - 12 PY - 2022 IS - 9 PG - 16 SN - 2073-4344 DO - 10.3390/catal12091065 UR - https://m2.mtmt.hu/api/publication/33099063 ID - 33099063 N1 - Funding Agency and Grant Number: Ministry for Innovation and Technology of Hungary from the National Research, Development and Innovation Fund [TKP2021-EGA-02]; National Research, Development and Innovation Office [SNN-125637]; Romanian Ministry of Education and Research, CCCDI-UEFISCDI within PNCDI III [PN-III-P2-2.1-PED-2019-5031] Funding text: The research reported in this paper is part of project no. TKP2021-EGA-02, implemented with the support provided by the Ministry for Innovation and Technology of Hungary from the National Research, Development and Innovation Fund, financed under the TKP2021 funding scheme. The National Research, Development and Innovation Office (Budapest, Hungary) is acknowledged for funding (SNN-125637). This work was also supported by a grant of the Romanian Ministry of Education and Research, CCCDI-UEFISCDI, project number PN-III-P2-2.1-PED-2019-5031, within PNCDI III. AB - This study implements a convenient microreactor for biocatalysis with enzymes immobilized on magnetic nanoparticles (MNPs). The enzyme immobilized onto MNPs by adsorption or by covalent bonds was lipase B from Candida antarctica (CaLB). The MNPs for adsorption were obtained by covering the magnetite core with a silica shell and later with hexadecyltrimethoxysilane, while for covalent immobilization, the silica-covered MNPs were functionalized by a layer forming from mixtures of hexadecyl- and 3-(2-aminoethylamino)propyldimethoxymethylsilanes in 16:1 molar ratio, which was further activated with neopentyl glycol diglycidyl ether (NGDE). The resulting CaLB-MNPs were tested in a convenient continuous flow system, created by 3D printing to hold six adjustable permanent magnets beneath a polytetrafluoroethylene tube (PTFE) to anchor the MNP biocatalyst inside the tube reactor. The anchored CaLB-MNPs formed reaction chambers in the tube for passing the fluid through and above the MNP biocatalysts, thus increasing the mixing during the fluid flow and resulting in enhanced activity of CaLB on MNPs. The enantiomer selective acylation of 4-(morpholin-4-yl)butan-2-ol (±)-1, being the chiral alcohol constituent of the mucolytic drug Fedrilate, was carried out by CaLB-MNPs in the U-shape reactor. The CaLB-MNPs in the U-shape reactor were compared in batch reactions to the lyophilized CaLB and to the CaLB-MNPs using the same reaction composition, and the same amounts of CaLB showed similar or higher activity in flow mode and superior activity as compared to the lyophilized powder form. The U-shape permanent magnet design represents a general and easy-to-access implementation of MNP-based flow microreactors, being useful for many biotransformations and reducing costly and time-consuming downstream processes. LA - English DB - MTMT ER - TY - JOUR AU - Nagy, Flóra AU - Bell, Evelin AU - Jipa, Monica AU - Hornyánszky, Gábor AU - Szilágyi, András Ferenc AU - Nagyné László, Krisztina AU - Katona, Gábor AU - Paizs, Csaba AU - Poppe, László AU - Balogh Weiser, Diána TI - Cross-Linked Enzyme-Adhered Nanoparticles (CLEANs) for Continuous-Flow Bioproduction JF - CHEMSUSCHEM J2 - CHEMSUSCHEM VL - 15 PY - 2022 IS - 2 PG - 10 SN - 1864-5631 DO - 10.1002/cssc.202102284 UR - https://m2.mtmt.hu/api/publication/32515084 ID - 32515084 N1 - Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest, 1111, Hungary Biocatalysis and Biotransformation Research Center, Babeş-Bolyai University of Cluj-Napoca, Arany János str. 11, Cluj-Napoca, 400028, Romania Department of Physical Chemistry and Materials Science, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest, 1111, Hungary SynBiocat LLC, Szilasliget u 3, Budapest, 1072, Hungary Export Date: 5 April 2022 CODEN: CHEMI Correspondence Address: Poppe, L.; Department of Organic Chemistry and Technology, Műegyetem rkp. 3, Hungary; email: poppe.laszlo@vbk.bme.hu Correspondence Address: Balogh-Weiser, D.; Department of Organic Chemistry and Technology, Műegyetem rkp. 3, Hungary; email: balogh.weiser.diana@vbk.bme.hu Funding details: 103413, P37_273 Funding details: 131467 Funding details: Budapesti Műszaki és Gazdaságtudományi Egyetem, BME Funding details: Magyar Tudományos Akadémia, MTA, BO/00175/21, ÚNKP‐21‐5‐BME‐386 Funding details: Emberi Eroforrások Minisztériuma, EMMI Funding text 1: This work was supported by the Higher Education Excellence Program of the Ministry of Human Capacities (Budapest, Hungary) in the frame of Biotechnology research area of Budapest University of Technology and Economics (TKP2020 IES, Grant No. BME‐IE‐BIO). D.B.W. thanks support form NKFIH‐PD‐19 (ID: 131467) and acknowledges the János Bolyai Research Scholarship of the Hungarian Academy of Sciences (BO/00175/21) and the ÚNKP‐21‐5 (ÚNKP‐21‐5‐BME‐386) New National Excellence Program of the Ministry of Human Capacities. C.P., G.K., and L.P. thank the support of NEMSyB, ID P37_273, Cod MySMIS 103413 [funded by Ministry of Education and Research, Bucharest, Romania and European Regional Development Fund, Competitiveness Operational Program 2014–2020 (POC), Priority axis 1, Action 1.1]. D.B.W. was also financed by CELSA (KU Leuven‐BME: ConSolid, 2020). AB - Nanostructured but micro-sized biocatalysts were created by bottom-up technology using multi-functionalized silica nanoparticles (NPs) as nano-sized building blocks to form cross-linked enzyme-adhered nanoparticles (CLEANs) as robust micro-sized particles with beneficial internal structure and good mechanical properties. Systematic surface modification of NPs with a grafting mixture consisting of organosilanes with reactive (aminopropyl) and inert (e. g., vinyl, propyl, phenyl, or octyl) functions resulted in functional NPs enabling cross-linking agents, such as glutardialdehyde or bisepoxides (glycerol diglycidyl ether, neopentylglycol diglycidyl ether, and poly(propylene glycol) diglycidyl ether), to bind and cross-link enzymes covalently and to form macroporous microparticles. These CLEANs were able to diminish several weaknesses of traditional cross-linked enzyme aggregates as biocatalysts, such as poor mechanical resistance, difficult recovery, and storage, strengthening their use for packed-bed enzyme reactors. Lipase B from Candida antarctica (CaLB) was selected as model enzyme for development of robust CLEANs, which were successfully tested for various industrially relevant applications including a kinetic resolution of a racemic alcohol and the production of various natural fragrance compounds under continuous-flow conditions. LA - English DB - MTMT ER - TY - JOUR AU - Koplányi, Gábor AU - Bell, Evelin AU - Molnár, Zsófia Klára AU - Tóth, Gergő Dániel AU - Józó, Muriel AU - Szilágyi, András Ferenc AU - Ender, Ferenc AU - Pukánszky, Béla AU - Vértessy, Beáta (Grolmuszné) AU - Poppe, László AU - Balogh Weiser, Diána TI - Entrapment of Phenylalanine Ammonia-Lyase in Nanofibrous Polylactic Acid Matrices by Emulsion Electrospinning JF - CATALYSTS J2 - CATALYSTS VL - 11 PY - 2021 IS - 10 PG - 14 SN - 2073-4344 DO - 10.3390/catal11101149 UR - https://m2.mtmt.hu/api/publication/32242806 ID - 32242806 N1 - Export Date: 26 October 2021 LA - English DB - MTMT ER - TY - THES AU - Bell, Evelin TI - Fém-fehérje komplex és átmenetifém katalizátorok fejlesztése és alkalmazása PB - Budapesti Műszaki és Gazdaságtudományi Egyetem PY - 2021 SP - 137 UR - https://m2.mtmt.hu/api/publication/32217259 ID - 32217259 LA - Hungarian DB - MTMT ER - TY - JOUR AU - Bell, Evelin AU - Kovacs, Norbert Krisztian AU - Alacs, Balint AU - Molnár, Zsófia Klára AU - Hornyánszky, Gábor TI - Immobilization of Phenylalanine Ammonia-lyase via EDTA Based Metal Chelate Complexes - Optimization and Prospects JF - PERIODICA POLYTECHNICA-CHEMICAL ENGINEERING J2 - PERIOD POLYTECH CHEM ENG VL - 65 PY - 2021 IS - 3 SP - 308 EP - 319 PG - 12 SN - 0324-5853 DO - 10.3311/PPch.17891 UR - https://m2.mtmt.hu/api/publication/32083731 ID - 32083731 N1 - Funding Agency and Grant Number: NRDI Fund [project (TKP2020 IES)] under Ministry for Innovation and Technology [BME-IE-BIO]; [SNN-125637] Funding text: The research reported in this paper and carried out at BME has been supported by the NRDI Fund [project (TKP2020 IES, Grant No. BME-IE-BIO) based on the charter of bolster issued by the NRDI Office under the auspices of the Ministry for Innovation and Technology; and project (SNN-125637)]. AB - 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 aminoaikyl moieties modified with an EDTA dianhydride (EDTADa)-derived chelator and charged with transition metal ions. Out of the five investigated metal ions - Fe3+, Co2+, NP2+, 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 (Y-A=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. LA - English DB - MTMT ER - TY - JOUR AU - Imarah, Ali O. AU - Csuka, Pál AU - Bataa, Naran AU - Decsi, Balázs AU - Bell, Evelin AU - Molnár, Zsófia Klára AU - Balogh Weiser, Diána AU - Poppe, László TI - Magnetically Agitated Nanoparticle-Based Batch Reactors for Biocatalysis with Immobilized Aspartate Ammonia-Lyase JF - CATALYSTS J2 - CATALYSTS VL - 11 PY - 2021 IS - 4 PG - 13 SN - 2073-4344 DO - 10.3390/catal11040483 UR - https://m2.mtmt.hu/api/publication/31959087 ID - 31959087 N1 - Funding Agency and Grant Number: National Research, Development and Innovation Office (Budapest, Hungary) [TKP2020 IES, BME-IE-BIO]; OTKAOrszagos Tudomanyos Kutatasi Alapprogramok (OTKA) [SNN-125637, PD-131467] Funding text: This research was funded by the National Research, Development and Innovation Office (Budapest, Hungary), grant numbers TKP2020 IES, Grant No. BME-IE-BIO, and OTKA, SNN-125637 and PD-131467. Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest, H-1111, Hungary Chemical Engineering Department, College of Engineering, University of Babylon, Hilla Babylon, 5100, Iraq Fermentia Microbiological Ltd, Berlini út 47-49, Budapest, H-1045, Hungary Institute of Enzymology, Research Center for Natural Sciences, Hungarian Academy of Science, Magyar tudósok körútja 2, Budapest, H-1117, Hungary Department of Phsysical Chemistry and Materials Science, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest, H-1111, Hungary SynBiocat Ltd, Szilasliget u 3, Budapest, H-1172, Hungary Biocatalysis and Biotransformation Research Center, Faculty of Chemistry and Chemical Engineering, Babeş-Bolyai University of Cluj-Napoca, Arany János Str. 11, Cluj-Napoca, RO-400028, Romania Export Date: 13 May 2021 Correspondence Address: Poppe, L.; Department of Organic Chemistry and Technology, Műegyetem rkp. 3, Hungary; email: poppe.laszlo@vbk.bme.hu Correspondence Address: Poppe, L.; SynBiocat Ltd, Szilasliget u 3, Hungary; email: poppe.laszlo@vbk.bme.hu Correspondence Address: Poppe, L.; Biocatalysis and Biotransformation Research Center, Arany János Str. 11, Romania; email: poppe.laszlo@vbk.bme.hu Funding details: Hungarian Scientific Research Fund, OTKA, PD-131467, SNN-125637 Funding details: National Research, Development and Innovation Office Funding text 1: Funding: This research was funded by the National Research, Development and Innovation Office (Budapest, Hungary), grant numbers TKP2020 IES, Grant No. BME-IE-BIO, and OTKA, SNN-125637 and PD-131467. Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest, H-1111, Hungary Chemical Engineering Department, College of Engineering, University of Babylon, Hilla Babylon, 5100, Iraq Fermentia Microbiological Ltd, Berlini út 47-49, Budapest, H-1045, Hungary Institute of Enzymology, Research Center for Natural Sciences, Hungarian Academy of Science, Magyar tudósok körútja 2, Budapest, H-1117, Hungary Department of Phsysical Chemistry and Materials Science, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest, H-1111, Hungary SynBiocat Ltd, Szilasliget u 3, Budapest, H-1172, Hungary Biocatalysis and Biotransformation Research Center, Faculty of Chemistry and Chemical Engineering, Babeş-Bolyai University of Cluj-Napoca, Arany János Str. 11, Cluj-Napoca, RO-400028, Romania Export Date: 14 May 2021 Correspondence Address: Poppe, L.; Department of Organic Chemistry and Technology, Műegyetem rkp. 3, Hungary; email: poppe.laszlo@vbk.bme.hu Correspondence Address: Poppe, L.; SynBiocat Ltd, Szilasliget u 3, Hungary; email: poppe.laszlo@vbk.bme.hu Correspondence Address: Poppe, L.; Biocatalysis and Biotransformation Research Center, Arany János Str. 11, Romania; email: poppe.laszlo@vbk.bme.hu Funding details: Hungarian Scientific Research Fund, OTKA, PD-131467, SNN-125637 Funding details: National Research, Development and Innovation Office Funding text 1: Funding: This research was funded by the National Research, Development and Innovation Office (Budapest, Hungary), grant numbers TKP2020 IES, Grant No. BME-IE-BIO, and OTKA, SNN-125637 and PD-131467. AB - In this study, we investigated the influence of different modes of magnetic mixing on effective enzyme activity of aspartate ammonia-lyase from Pseudomonas fluorescens immobilized onto epoxy-functionalized magnetic nanoparticles by covalent binding (AAL-MNP). The effective specific enzyme activity of AAL-MNPs in traditional shake vial method was compared to the specific activity of the MNP-based biocatalyst in two devices designed for magnetic agitation. The first device agitated the AAL-MNPs by moving two permanent magnets at two opposite sides of a vial in x-axis direction (being perpendicular to the y-axis of the vial); the second device unsettled the MNP biocatalyst by rotating the two permanent magnets around the y-axis of the vial. In a traditional shake vial, the substrate and biocatalyst move in the same direction with the same pattern. In magnetic agitation modes, the MNPs responded differently to the external magnetic field of two permanent magnets. In the axial agitation mode, MNPs formed a moving cloud inside the vial, whereas in the rotating agitation mode, they formed a ring. Especially, the rotating agitation of the MNPs generated small fluid flow inside the vial enabling the mixing of the reaction mixture, leading to enhanced effective activity of AAL-MNPs compared to shake vial agitation. LA - English DB - MTMT ER - TY - JOUR AU - Bata, Zsófia AU - Molnár, Zsófia Klára AU - Madaras, Erzsébet AU - Molnár, Bence AU - Bell, Evelin AU - Varga, Andrea AU - Leveles, Ibolya AU - Qian, Renzhe AU - Hammerschmidt, Friedrich AU - Paizs, Csaba AU - Vértessy, Beáta (Grolmuszné) AU - Poppe, László TI - Substrate Tunnel Engineering Aided by X-ray Crystallography and Functional Dynamics Swaps the Function of MIO-Enzymes JF - ACS CATALYSIS J2 - ACS CATAL VL - 11 PY - 2021 IS - 8 SP - 4538 EP - 4549 PG - 12 SN - 2155-5435 DO - 10.1021/acscatal.1c00266 UR - https://m2.mtmt.hu/api/publication/31939010 ID - 31939010 AB - The enzyme family harboring the post-translationally formed 5-methylene-3,5-dihydro-4H-imidazol-4-one (MIO) catalytic residue comprises both aromatic amino acid ammonia-lyases (ALs) and 2,3-aminomutases (AMs). The structural origin of the different functions and the role of the inner loop region in substrate binding are not fully understood. Here, we provide the three-dimensional structures for Petroselinum crispum phenylalanine AL (PcPAL) with fully resolved inner loops in a catalytically competent conformation. Using molecular modeling, we demonstrate that in both ALs and AMs of eukaryotic origin, just a small opening of the inner loop is sufficient for ligand egress. Furthermore, we show that ligand-binding tunnels are analogous to eukaryotic MIO-enzymes and that the critical initial part of these tunnels is present across the whole enzyme family. Engineering of these binding tunnels converts an (R)-AM to a highly selective (S)-β-AL thus establishing a nonclassified enzyme function. LA - English DB - MTMT ER - TY - JOUR AU - Bell, Evelin AU - Molnár, Zsófia Klára AU - Varga, Andrea AU - Nagy, Flóra AU - Hornyánszky, Gábor AU - Paizs, Csaba AU - Balogh Weiser, Diána AU - Poppe, László TI - “Fishing and hunting”– Selective immobilization of a recombinant phenylalanine ammonia-lyase from fermentation media JF - MOLECULES J2 - MOLECULES VL - 24 PY - 2019 IS - 22 SN - 1420-3049 DO - 10.3390/molecules24224146 UR - https://m2.mtmt.hu/api/publication/30920179 ID - 30920179 N1 - Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Budapest, 1111, Hungary Fermentia Microbiological Ltd., Budapest, 1405, Hungary Institute of Enzymology, Research Center for Natural Sciences, Hungarian Academy of Science, Budapest, 1117, Hungary Biocatalysis and Biotransformation Research Centre, Faculty of Chemistry and Chemical Engineering, Babeş-Bolyai University of Cluj-Napoca, Cluj-Napoca, 400028, Romania SynBiocat Ltd., Budapest, 1172, Hungary Department of Physical Chemistry and Materials Science, Budapest University of Technology and Economics, Budapest, 1111, Hungary Cited By :7 Export Date: 6 April 2022 CODEN: MOLEF Correspondence Address: Balogh-Weiser, D.; Department of Organic Chemistry and Technology, Hungary; email: dweiser@mail.bme.hu Chemicals/CAS: phenylalanine ammonia lyase, 9024-28-6; Enzymes, Immobilized; Phenylalanine Ammonia-Lyase; Plant Proteins; Recombinant Proteins Funding details: NTP-NFTÖ-18-B-0379 Funding details: Budapesti Műszaki és Gazdaságtudományi Egyetem, BME Funding details: National Authority for Scientific Research and Innovation, ANCSI Funding details: European Cooperation in Science and Technology, COST, 103413, CM 1303, P37_273 Funding details: Emberi Eroforrások Minisztériuma, EMMI Funding details: European Regional Development Fund, FEDER Funding details: Nemzeti Kutatási Fejlesztési és Innovációs Hivatal, NKFIH, SNN-125637 Funding text 1: Acknowledgments: This work was supported by National Talent Program of Ministry of Human Capacities (NTP-NFTÖ-18-B-0379). Funding text 2: This work was supported NKFI (Budapest, Hungary; SNN-125637) and by the Higher Education Excellence Program of the Ministry of Human Capacities (Budapest, Hungary) in the frame of Biotechnology research area of Budapest University of Technology and Economics (BME FIKP-BIO). CP and LP thank the COST Action SysBiocat (CM 1303) and project NEMSyB, ID P37-273, Cod MySMIS 103413 [funded by National Authority for Scientific Research and Innovation (ANCSI, Bucharest, Romania) and European Regional Development Fund, Competitiveness Operational Program 2014-2020 (POC), Priority axis 1, Action 1.1] for support. Funding text 3: Funding: This work was supported NKFI (Budapest, Hungary; SNN-125637) and by the Higher Education Excellence Program of the Ministry of Human Capacities (Budapest, Hungary) in the frame of Biotechnology research area of Budapest University of Technology and Economics (BME FIKP-BIO). CP and LP thank the COST Action SysBiocat (CM 1303) and project NEMSyB, ID P37_273, Cod MySMIS 103413 [funded by National Authority for Scientific Research and Innovation (ANCSI, Bucharest, Romania) and European Regional Development Fund, Competitiveness Operational Program 2014-2020 (POC), Priority axis 1, Action 1.1] for support. AB - 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. LA - English DB - MTMT ER -