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

Dima, Norbert A. and Filip, Alina and Bencze, László Csaba and Oláh, Márk and Sátorhelyi, Péter and Vértessy, Beáta (Grolmuszné) and Poppe, László (2016) Expression and purification of recombinant phenylalanine ammonia-lyase from Petroselinum crispum. STUDIA UNIVERSITATIS BABES-BOLYAI CHEMIA, 61 (2). pp. 21-34. ISSN 1224-7154

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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.

Item Type: Article
Subjects: Q Science / természettudomány > QD Chemistry / kémia
Q Science / természettudomány > QH Natural history / természetrajz > QH301 Biology / biológia > QH3011 Biochemistry / biokémia
T Technology / alkalmazott, műszaki tudományok > TP Chemical technology / vegyipar, vegyészeti technológia
SWORD Depositor: MTMT SWORD
Depositing User: MTMT SWORD
Date Deposited: 24 Jan 2017 08:13
Last Modified: 24 Jan 2017 08:13
URI: http://real.mtak.hu/id/eprint/46153

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