Zymobacter palmae pyruvate decarboxylase production process development : Cloning in Escherichia coli, fed-batch culture and purification
| Autor: | Natàlia Alcover, Marina Guillén, Albert Carceller, Gregorio Álvaro |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Alanine
Environmental Engineering Downstream processing Chemistry Ion chromatography High cell density cultures Bioengineering Fast protein liquid chromatography hemic and immune systems Pyruvate decarboxylase (PDC) macromolecular substances medicine.disease_cause PDC purification Fed-batch culture Biochemistry medicine Bioreactor Zymobacter palmae Escherichia coli Research Articles Pyruvate decarboxylase Biotechnology |
| Zdroj: | Dipòsit Digital de Documents de la UAB Universitat Autònoma de Barcelona Recercat: Dipósit de la Recerca de Catalunya Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya) Eng Life Sci Recercat. Dipósit de la Recerca de Catalunya instname |
| Popis: | 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(−1)h(−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. |
| Databáze: | OpenAIRE |
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