Zymobacter palmae pyruvate decarboxylase production process development: Cloning in Escherichia coli, fed-batch culture and purification

Natàlia Alcover, Albert Carceller, Gregorio Álvaro, Marina Guillén

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© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim 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.
Idioma originalAnglès
Pàgines (de-a)502-512
Nombre de pàgines11
RevistaEngineering in Life Sciences
Volum19
Número7
DOIs
Estat de la publicacióPublicada - 1 de jul. 2019

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