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Fusion of a Coiled‐Coil Domain Facilitates the High‐Level Production of Catalytically Active Enzyme Inclusion Bodies
Author(s) -
Diener Martin,
Kopka Benita,
Pohl Martina,
Jaeger KarlErich,
Krauss Ulrich
Publication year - 2016
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201501001
Subject(s) - bacillus subtilis , chemistry , enzyme , lipase , escherichia coli , aqueous solution , inclusion bodies , combinatorial chemistry , organic chemistry , biochemistry , biology , bacteria , gene , genetics
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 ( Bs LA), the hydroxynitrile lyase of Arabidopsis thaliana ( At HNL), and the 2‐succinyl‐5‐enolpyruvyl‐6‐hydroxy‐3‐cyclohexene‐1‐carboxylate synthase MenD of E. coli ( Ec MenD) were successfully produced as CatIBs and used in aqueous and micro‐aqueous organic solvent based reaction systems, showing excellent stability and recyclability.