Biosensor-Guided Engineering of a Baeyer-Villiger Monooxygenase for Aliphatic Ester Production

Overview

Journal Chembiochem

Specialty Biochemistry

Date 2024 Sep 25

PMID 39320950

Authors

Thaleia Sakoleva

Florian Vesenmaier

Lena Koch

Jarne E Schunke

Kay D Novak

Sascha Grobe

Mark Dorr

Uwe T Bornscheuer

Thomas Bayer

Affiliations

Soon will be listed here.

Abstract

Esters are valuable aroma compounds and can be produced enzymatically by Baeyer-Villiger monooxygenases (BVMOs) from (aliphatic) ketone precursors. However, a genetically encoded biosensor system for the assessment of BVMO activity and the detection of reaction products is missing. In this work, we assembled a synthetic enzyme cascade - featuring an esterase, an alcohol dehydrogenase, and LuxAB - in the heterologous host Escherichia coli. Target esters are produced by a BVMO, subsequently cleaved, and the corresponding alcohol oxidized through the artificial pathway. Ultimately, aldehyde products are detected in vivo by LuxAB, a luciferase from Photorhabdus luminescens that emits bioluminescence upon the oxidation of aldehydes to the corresponding carboxylates. This biosensor system greatly accelerated the screening and selection of active BVMO variants from a focused library, omitting commonly used low-throughput chromatographic analysis. Engineered enzymes accepted linear aliphatic ketones such as 2-undecanone and 2-dodecanone and exhibited improved ester formation.

Citing Articles

Biosensor-Guided Engineering of a Baeyer-Villiger Monooxygenase for Aliphatic Ester Production.

Sakoleva T, Vesenmaier F, Koch L, Schunke J, Novak K, Grobe S Chembiochem. 2024; 26(1):e202400712.

PMID: 39320950 PMC: 11727011. DOI: 10.1002/cbic.202400712.

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