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Licensed Unlicensed Requires Authentication Published by De Gruyter June 10, 2016

Fusion proteins of an enoate reductase and a Baeyer-Villiger monooxygenase facilitate the synthesis of chiral lactones

  • Christin Peters , Florian Rudroff , Marko D. Mihovilovic and Uwe T. Bornscheuer EMAIL logo
From the journal Biological Chemistry


Nature uses the advantages of fusion proteins for multi-step reactions to facilitate the metabolism in cells as the conversion of substrates through intermediates to the final product can take place more rapidly and with less side-product formation. In a similar fashion, also for enzyme cascade reactions, the fusion of biocatalysts involved can be advantageous. In the present study, we investigated fusion of an alcohol dehydrogenase (ADH), an enoate reductase (ERED) and a Baeyer-Villiger monooxygenase (BVMO) to enable the synthesis of (chiral) lactones starting from unsaturated alcohols as substrates. The domain order and various linkers were studied to find optimal conditions with respect to expression levels and enzymatic activities. Best results were achieved for the ERED xenobiotic reductase B (XenB) from Pseudomonas putida and the cyclohexanone monooxygenase (CHMO) from Acinetobacter sp., whereas none of the ADHs studied could be fused successfully. This fusion protein together with separately supplied ADH resulted in similar reaction rates in in vivo biocatalysis reactions. After 1.5 h we could detect 40% more dihydrocarvone lactone in in vivo reactions with the fusion protein and ADH then with the single enzymes.

Dedicated to Professor Romas J. Kazlauskas on the occasion of his 60th birthday.


We thank the DFG (grant no Bo1862/6-1) and the FWF (grant no. I723-N17) for financial support.


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Supplemental Material:

The online version of this article (DOI: 10.1515/hsz-2016-0150) offers supplementary material, available to authorized users.

Received: 2016-3-15
Accepted: 2016-6-8
Published Online: 2016-6-10
Published in Print: 2017-1-1

©2017 Walter de Gruyter GmbH, Berlin/Boston

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