Zymobacter palmae pyruvate decarboxylase production process development : Cloning in Escherichia coli, fed-batch culture and purification
Alcover Fortuny, Natàlia (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Carceller, Albert (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Álvaro, Gregorio (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Guillén, Marina (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)

Date:	2019
Abstract:	Pyruvate decarboxylase (PDC) is responsible for the decarboxylation of pyruvate, producing acetaldehyde and carbon dioxide and is of high interest for industrial applications. PDC is a very powerful tool in the enzymatic synthesis of chiral amines by combining it with transaminases when alanine is used as amine donor. However, one of the main drawback that hampers its use in biocatalysis is its production and the downstream processing on scale. In this paper, a production process of PDC from Zymobacter palmae has been developed. The enzyme has been cloned and overexpressed in Escherichia coli. It is presented, for the first time, the evaluation of the production of recombinant PDC in a bench-scale bioreactor, applying a substrate-limiting fed-batch strategy which led to a volumetric productivity and a final PDC specific activity of 6942 U L−1h−1 and 3677 U gDCW−1 (dry cell weight). Finally, PDC was purified in fast protein liquid chromatography equipment by ion exchange chromatography. The developed purification process resulted in 100% purification yield and a purification factor of 3. 8.
Grants:	Ministerio de Economía y Competitividad CTQ2014-53114-R Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-1462 Ministerio de Economía y Competitividad BES-2015-072542
Rights:	Tots els drets reservats.
Language:	Anglès
Document:	Article ; recerca ; Versió acceptada per publicar
Subject:	Fed-batch culture ; High cell density cultures ; PDC purification ; Pyruvate decarboxylase (PDC) ; Zymobacter palmae
Published in:	Engineering in life science, Vol. 19, Issue 7 (July 2019) , p. 502-512, ISSN 1618-0240

DOI: 10.1002/elsc.201900010
PMID: 32625027

Postprint 33 p, 1.6 MB

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