Expression and purification of recombinant phenylalanine ammonia-lyase from Petroselinum crispum

Norbert Artur Dima, Alina Filip, László Csaba Bencze, Márk Oláh, Péter Sátorhelyi, B. Vértessy, L. Poppe, Csaba Paizs

Research output: Contribution to journalArticle

8 Citations (Scopus)

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 KM, and kcat values suggesting excellent catalytic properties of the recombinant enzyme.

Original languageEnglish
Pages (from-to)21-34
Number of pages14
JournalStudia Universitatis Babes-Bolyai Chemia
Volume61
Issue number2
Publication statusPublished - 2016

Fingerprint

Phenylalanine Ammonia-Lyase
Recombinant Proteins
Purification
Cloning
Escherichia coli
Genes
Substrates
Enzymes
Proteins

Keywords

  • Expression and expression optimization
  • Molecular cloning
  • Petroselinum crispum
  • Phenylalanine ammonia-lyase

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Dima, N. A., Filip, A., Bencze, L. C., Oláh, M., Sátorhelyi, P., Vértessy, B., ... Paizs, C. (2016). Expression and purification of recombinant phenylalanine ammonia-lyase from Petroselinum crispum. Studia Universitatis Babes-Bolyai Chemia, 61(2), 21-34.

Expression and purification of recombinant phenylalanine ammonia-lyase from Petroselinum crispum. / Dima, Norbert Artur; Filip, Alina; Bencze, László Csaba; Oláh, Márk; Sátorhelyi, Péter; Vértessy, B.; Poppe, L.; Paizs, Csaba.

In: Studia Universitatis Babes-Bolyai Chemia, Vol. 61, No. 2, 2016, p. 21-34.

Research output: Contribution to journalArticle

Dima, Norbert Artur ; Filip, Alina ; Bencze, László Csaba ; Oláh, Márk ; Sátorhelyi, Péter ; Vértessy, B. ; Poppe, L. ; Paizs, Csaba. / Expression and purification of recombinant phenylalanine ammonia-lyase from Petroselinum crispum. In: Studia Universitatis Babes-Bolyai Chemia. 2016 ; Vol. 61, No. 2. pp. 21-34.
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