[Zurück]

C. Peters, F. Rudroff, M.D. Mihovilovic, U. Bornscheuer:
"A fusion protein of an enoate reductase and a Baeyer-Villiger monoxygenase facilitates synthesis of chiral lactones";
Biological Chemistry, 1 (2017), 398; S. 31 - 37.

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 sideproduct
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.

alcohol dehydrogenase; Baeyer-Villiger monooxygenase; biocatalysis; enoate reductase; enzyme cascade; fusion proteins

http://dx.doi.org/10.1515/hsz-2016-0150