Fusion of a Coiled-Coil Domain Facilitates the High-Level Production of Catalytically Active Enzyme Inclusion Bodies

The increasing number of biocatalytic reactions implemented in chemical synthesis routes raises the urgent need for large amounts of enzymes. Hence, new generic methods are required for their simple and cost‐efficient production. Here, we describe a generally applicable method based on the productio...

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Bibliographic Details
Published in:ChemCatChem 2016-01, Vol.8 (1), p.142-152
Main Authors: Diener, Martin, Kopka, Benita, Pohl, Martina, Jaeger, Karl-Erich, Krauss, Ulrich
Format: Article
Language:English
Subjects:
Bacteriology
biocatalysis
enzyme immobilizates
Enzymes
inclusion bodies
organic media
synthetic chemistry
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Summary:The increasing number of biocatalytic reactions implemented in chemical synthesis routes raises the urgent need for large amounts of enzymes. Hence, new generic methods are required for their simple and cost‐efficient production. Here, we describe a generally applicable method based on the production of catalytically active inclusion bodies (CatIBs). CatIBs represent a promising new form of biologically produced, carrier‐free, biodegradable enzyme immobilizate. CatIBs are produced in Escherichia coli by expression of a gene fusion consisting of a coiled‐coil domain and a target enzyme. Employing this strategy, the lipase A of Bacillus subtilis (BsLA), the hydroxynitrile lyase of Arabidopsis thaliana (AtHNL), and the 2‐succinyl‐5‐enolpyruvyl‐6‐hydroxy‐3‐cyclohexene‐1‐carboxylate synthase MenD of E. coli (EcMenD) were successfully produced as CatIBs and used in aqueous and micro‐aqueous organic solvent based reaction systems, showing excellent stability and recyclability. Fusion of a coiled‐coil domain to various enzymes results in the formation of catalytically active inclusion bodies (CatIBs). These represent a novel type of biologically produced, biodegradable, and carrier‐free enzyme immobilizate for applications in lab‐scale biocatalysis research, synthetic chemistry, and industry.
ISSN:1867-3880
1867-3899
DOI:10.1002/cctc.201501001