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Issue 22, 2018
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Tailor-made catalytically active inclusion bodies for different applications in biocatalysis

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Abstract

We have recently demonstrated that fusions of different target enzymes to the coiled-coil domain TDoT induced the formation of catalytically active inclusion bodies (CatIBs) in E. coli (doi:10.1016/j.jbiotec.2017.04.033). Here we show that the CatIB properties can be tailored to the requirements of different reaction systems using two different coiled-coil domains as fusion tags: TDoT and 3HAMP. As an example the benzaldehyde lyase from Pseudomonas fluorescens (PfBAL) was chosen, which catalyzes the formation of 2-hydroxy ketones and benzoins from aromatic donor aldehydes and aromatic or aliphatic acceptor aldehydes. Using these fusion tags two different kinds of PfBAL-CatIBs were successfully produced that differ in morphology, solubility during washing steps, initial rate activity, protein and lipid content. TDoT-PfBAL and 3HAMP-PfBAL CatIBs were studied relative to the soluble enzyme concerning their application in biocatalysis in continuous reaction mode and in batch using a mono- and biphasic solvent system. Both CatIBs outperformed the soluble enzyme with respect to stability under reaction conditions. In the buffer/DMSO system used for the continuous synthesis of (R)-2-hydroxy-1-phenylpropanone, TDoT-PfBAL was superior to the soluble enzyme. Besides, 3HAMP-PfBAL was more suitable in a biphasic reaction system for the synthesis of (R)-benzoins and revealed an up to 3-fold higher activity compared to the soluble enzyme.

Graphical abstract: Tailor-made catalytically active inclusion bodies for different applications in biocatalysis

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Supplementary files

Article information


Submitted
10 Sep 2018
Accepted
12 Oct 2018
First published
15 Oct 2018

Catal. Sci. Technol., 2018,8, 5816-5826
Article type
Paper

Tailor-made catalytically active inclusion bodies for different applications in biocatalysis

R. Kloss, T. Karmainski, V. D. Jäger, D. Hahn, A. Grünberger, M. Baumgart, U. Krauss, K. Jaeger, W. Wiechert and M. Pohl, Catal. Sci. Technol., 2018, 8, 5816
DOI: 10.1039/C8CY01891J

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