Novel Approach for the Isolation and Immobilization of a Recombinant Transaminase: Applying an Advanced Nanocomposite System

Gábor Koplányi; Evelin Bell; Zsófia Molnár; Gábor Katona; Péter Lajos Neumann; Ferenc Ender; György T Balogh; Polona Žnidaršič-Plazl; László Poppe; Diána Balogh-Weiser

doi:10.1002/cbic.202200713

Novel Approach for the Isolation and Immobilization of a Recombinant Transaminase: Applying an Advanced Nanocomposite System

Chembiochem. 2023 Apr 3;24(7):e202200713. doi: 10.1002/cbic.202200713. Epub 2023 Mar 2.

Authors

Gábor Koplányi¹, Evelin Bell¹, Zsófia Molnár^{1

2}, Gábor Katona³, Péter Lajos Neumann^{4

5}, Ferenc Ender^{4

6}, György T Balogh^{7

8}, Polona Žnidaršič-Plazl⁹, László Poppe^{1

10}, Diána Balogh-Weiser^{1

11}

Affiliations

¹ Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, 1111, Műegyetem rkp. 3., Budapest, Hungary.
² Institute of Enzymology, ELKH Research Center of Natural Sciences, 1117, Magyar tudosók krt. 2. Budapest, Hungary.
³ Institute of Pharmaceutical Technology and Regulatory Affairs, University of Szeged, 6720, Eötvös u. 6., Szeged, Hungary.
⁴ Department of Electron Devices, Budapest University of Technology and Economics, 1111, Műegyetem rkp. 3., Budapest, Hungary.
⁵ Centre for Energy Research, Institute for Technical Physics and Materials Science, 1121, Konkoly-Thege M. út 29-33., Budapest, Hungary.
⁶ SpinSplit Llc., 1025, Vend u. 17., Budapest, Hungary.
⁷ Department of Chemical and Environmental Process Engineering, Budapest University of Technology and Economics, 1111, Műegyetem rkp. 3., Budapest, Hungary.
⁸ Institute of Pharmacodynamics and Biopharmacy, University of Szeged, 6720, Eötvös u. 6., Szeged, Hungary.
⁹ Faculty of Chemistry and Chemical Technology, University of Ljubljana Večna pot 113., 1000, Ljubljana, Slovenia.
¹⁰ Biocatalysis and Biotransformation Research Center Faculty of Chemistry and Chemical Engineering, Babeş-Bolyai University of Cluj-Napoca, 400028, Arany János Str. 11, Cluj-Napoca, Romania.
¹¹ Department of Physical Chemistry and Materials Science, Budapest University of Technology and Economics, 1111, Műegyetem rkp. 3., Budapest, Hungary.

PMID: 36653306
DOI: 10.1002/cbic.202200713

Abstract

The increasing application of recombinant enzymes demands not only effective and sustainable fermentation, but also highly efficient downstream processing and further stabilization of the enzymes by immobilization. In this study, a novel approach for the isolation and immobilization of His-tagged transaminase from Chromobacterium violaceum (CvTA) has been developed. A recombinant of CvTA was simultaneously isolated and immobilized by binding on silica nanoparticles (SNPs) with metal affinity linkers and additionally within poly(lactic acid) (PLA) nanofibers. The linker length and the nature of the metal ion significantly affected the enzyme binding efficiency and biocatalytic activity of CvTA-SNPs. The formation of PLA nanofibers by electrospinning enabled rapid embedding of CvTA-SNPs biocatalysts and ensured enhanced stability and activity. The developed advanced immobilization method reduces the time required for enzyme isolation, purification and immobilization by more than fourfold compared to a classical stepwise technique.

Keywords: electrospinning; immobilization; nanocomposites; nanoparticles; transaminases.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Enzymes, Immobilized* / metabolism
Lipase
Metals
Nanocomposites*
Polyesters
Transaminases

Substances

Enzymes, Immobilized
Transaminases
Polyesters
Lipase
Metals