EXPRESSION AND PURIFICATION OF RECOMBINANT PHENYLALANINE AMMONIA-LYASE FROM PETROSELINUM CRISPUM

Norbert Artur  DIMA; Alina  FILIP; László Csaba BENCZE; Márk  OLÁH; Péter  SÁTORHELYI; Beáta G. VÉRTESSY; László POPPE; Csaba PAIZS

EXPRESSION AND PURIFICATION OF RECOMBINANT PHENYLALANINE AMMONIA-LYASE FROM PETROSELINUM CRISPUM

Authors

Norbert Artur DIMA Faculty of Chemistry and Chemical Engineering, Babeş-Bolyai University, Cluj-Napoca, Romania. Corresponding author: csaba.paizs@ubbcluj.ro.
Alina FILIP Research Center for Enzymology and Applied Biocatalysis, Faculty of Chemistry and Chemical Engineering, Babeş-Bolyai University, Cluj-Napoca, Romania. Emaill: alina.filip@ubbcluj.ro. https://orcid.org/0000-0001-6820-9204
László Csaba BENCZE Biocatalysis and Biotransformations Research Center, Babeş-Bolyai University, Faculty of Chemistry and Chemical Engineering, Cluj-Napoca, Romania. Email: cslbencze@chem.ubbcluj.ro. https://orcid.org/0000-0003-0956-9749
Márk OLÁH Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Hungary. Corresponding author: csaba.paizs@ubbcluj.ro. https://orcid.org/0000-0003-4628-6887
Péter SÁTORHELYI Fermentia Ltd, Budapest, Hungary. Corresponding author: paizs@chem.ubbcluj.ro.
Beáta G. VÉRTESSY Department of Biotechnology and Food Sciences, Budapest University of Technology and Economics; Institute of Enzymology, Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Budapest, Hungary. Email: vertessy.beata@ttk.mta.hu. https://orcid.org/0000-0002-1288-2982
László POPPE Department of Organic Chemistry and Technology, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Hungary. Email: poppe.laszlo@vbk.bme.hu. https://orcid.org/0000-0002-8358-1378
Csaba PAIZS Faculty of Chemistry and Chemical Engineering, Hungarian Line of Study, Babes-Bolyai University, Cluj Napoca, Romania. Email: csaba.paizs@ubbcluj.ro. https://orcid.org/0000-0002-7403-7098

Keywords:

phenylalanine ammonia-lyase, Petroselinum crispum, molecular cloning, expression and expression optimization

Abstract

In the present study the molecular cloning, expression and purification of recombinant PcPAL, with a cleavable N-terminal His-tag is described. The PcPAL gene was cloned into pET-19b vector and transformed to different E.coli host cells. The optimization of expression and purification processes provided recombinant protein with high purity in its native, tetrameric fold with a yield of 7-8 mg protein / 1 L culture. The activity of the recombinant protein was tested towards its natural substrate L-Phe, the K_M, and k_cat values suggesting excellent catalytic properties of the recombinant enzyme.

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STUDIA UBB CHEMIA, Volume 61 (LXI), No. 2, June 2016

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Published

2016-06-30

How to Cite

DIMA, N. A. ., FILIP, A. ., BENCZE, L. C., OLÁH, M. ., SÁTORHELYI, P. ., VÉRTESSY, B. G., … PAIZS, C. (2016). EXPRESSION AND PURIFICATION OF RECOMBINANT PHENYLALANINE AMMONIA-LYASE FROM PETROSELINUM CRISPUM. Studia Universitatis Babeș-Bolyai Chemia, 61(2), 21–34. Retrieved from https://studia.reviste.ubbcluj.ro/index.php/chemia/article/view/8305