Expression and purification of recombinant phenylalanine 2,3-aminomutase from Pantoea agglomerans

Andrea Varga, Alina Filip, László Csaba Bencze, Péter Sátorhelyi, Evelin Bell, B. Vértessy, L. Poppe, Csaba Paizs

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

In the present study, the gene of phenylalanine 2,3-aminomutase from Pantoea agglomerans (PaPAM) was cloned into pET-19b vector and used for its expression in competent Escherichia coli cells. The recombinant plasmid, PaPAM-pET-19b, was transformed into competent E. coli strain BL21(DE3)pLysS cells. Overnight culture of the transformed bacteria was induced by the addition of isopropylthio-β-D-galactoside (IPTG) to the final concentrations of 0.1, 0.5 and 1 mM. Also, the effects of different temperatures (18, 25 and 30°C) and the incubation time of PaPAM were examined. The fermentation process was scaled up to 10 L fermentor. Affinity purification conditions were analyzed by SDS-PAGE. The Tm and the activity of the purified enzyme was also investigated.

Original languageEnglish
Pages (from-to)7-19
Number of pages13
JournalStudia Universitatis Babes-Bolyai Chemia
Volume61
Issue number2
Publication statusPublished - 2016

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Phenylalanine
Purification
Escherichia coli
Galactosides
Fermentation
Bacteria
Plasmids
Genes
Enzymes
Temperature

Keywords

  • Optimization
  • Pantoea agglomerans
  • Phenylalanine 2,3-aminomutase
  • Protein expression

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Varga, A., Filip, A., Bencze, L. C., Sátorhelyi, P., Bell, E., Vértessy, B., ... Paizs, C. (2016). Expression and purification of recombinant phenylalanine 2,3-aminomutase from Pantoea agglomerans. Studia Universitatis Babes-Bolyai Chemia, 61(2), 7-19.

Expression and purification of recombinant phenylalanine 2,3-aminomutase from Pantoea agglomerans. / Varga, Andrea; Filip, Alina; Bencze, László Csaba; Sátorhelyi, Péter; Bell, Evelin; Vértessy, B.; Poppe, L.; Paizs, Csaba.

In: Studia Universitatis Babes-Bolyai Chemia, Vol. 61, No. 2, 2016, p. 7-19.

Research output: Contribution to journalArticle

Varga, Andrea ; Filip, Alina ; Bencze, László Csaba ; Sátorhelyi, Péter ; Bell, Evelin ; Vértessy, B. ; Poppe, L. ; Paizs, Csaba. / Expression and purification of recombinant phenylalanine 2,3-aminomutase from Pantoea agglomerans. In: Studia Universitatis Babes-Bolyai Chemia. 2016 ; Vol. 61, No. 2. pp. 7-19.
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