State-of-the-art in engineering small molecule biosensors and their applications in metabolic engineering

Chaisupa, Patarasuda; Wright, R. Clay

doi:10.1016/j.slast.2023.10.005

SLAS Technology

2024

DOI: 10.1016/j.slast.2023.10.005

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State-of-the-art in engineering small molecule biosensors and their applications in metabolic engineering

Patarasuda Chaisupa,

R. Clay Wright

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2024

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Cited by 3 publications

References 277 publications

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

Sakoleva,

Vesenmaier,

Koch

et al. 2024

ChemBioChem

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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.

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

Sakoleva,

Vesenmaier,

Koch

et al. 2024

ChemBioChem

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Genetically encoded biosensors for the circular plastics bioeconomy

Chacón,

Dixon

2024

Metabolic Engineering Communications

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Genetically Encoded, Noise-Tolerant, Auxin Biosensors in Yeast

Chaisupa,

Rahman,

Hildreth

et al. 2024

ACS Synth. Biol.

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Auxins are crucial signaling molecules that regulate the growth, metabolism, and behavior of various organisms, most notably plants but also bacteria, fungi, and animals. Many microbes synthesize and perceive auxins, primarily indole-3-acetic acid (IAA, referred to as auxin herein), the most prevalent natural auxin, which influences their ability to colonize plants and animals. Understanding auxin biosynthesis and signaling in fungi may allow us to better control interkingdom relationships and microbiomes from agricultural soils to the human gut. Despite this importance, a biological tool for measuring auxin with high spatial and temporal resolution has not been engineered in fungi. In this study, we present a suite of genetically encoded, ratiometric, protein-based auxin biosensors designed for the model yeast Saccharomyces cerevisiae. Inspired by auxin signaling in plants, the ratiometric nature of these biosensors enhances the precision of auxin concentration measurements by minimizing clonal and growth phase variation. We used these biosensors to measure auxin production across diverse growth conditions and phases in yeast cultures and calibrated their responses to physiologically relevant levels of auxin. Future work will aim to improve the fold change and reversibility of these biosensors. These genetically encoded auxin biosensors are valuable tools for investigating auxin biosynthesis and signaling in S. cerevisiae and potentially other yeast and fungi and will also advance quantitative functional studies of the plant auxin perception machinery, from which they are built.

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State-of-the-art in engineering small molecule biosensors and their applications in metabolic engineering

Cited by 3 publications

References 277 publications

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

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

Genetically encoded biosensors for the circular plastics bioeconomy

Genetically Encoded, Noise-Tolerant, Auxin Biosensors in Yeast

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