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A curcumin direct protein biosensor for cell-free prototyping

Kennedy, Agata, Griffin, Guy, Freemont, Paul S, Polizzi, Karen M, Moore, Simon J. (2022) A curcumin direct protein biosensor for cell-free prototyping. Engineering Biology, 6 (2-3). pp. 62-68. ISSN 2398-6182. (doi:10.1049/enb2.12024) (KAR id:100820)

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Official URL:
https://doi.org/10.1049/enb2.12024

Abstract

In synthetic biology, biosensors are routinely coupled with a gene expression system for detecting small molecules and physical signals. We reveal a fluorescent complex, based on the interaction of an coli double bond reductase ( CurA), as a detection unit with its substrate curcumin-we call this a direct protein (DiPro) biosensor. Using a cell-free synthetic biology approach, we use the CurA DiPro biosensor to fine tune 10 reaction parameters (cofactor, substrate, and enzyme levels) for cell-free curcumin biosynthesis, assisted through acoustic liquid handling robotics. Overall, we increase CurA-curcumin DiPro fluorescence within cell-free reactions by 78-fold. This finding adds to the growing family of protein-ligand complexes that are naturally fluorescent and potentially exploitable for a range of applications, including medical imaging to engineering high-value chemicals.

Item Type: Article
DOI/Identification number: 10.1049/enb2.12024
Uncontrolled keywords: curcumin, synthetic biology, fine chemical, cell‐free, biosensor
Subjects: Q Science > QR Microbiology
Divisions: Divisions > Division of Natural Sciences > Biosciences
Funders: Engineering and Physical Sciences Research Council (https://ror.org/0439y7842)
SWORD Depositor: JISC Publications Router
Depositing User: JISC Publications Router
Date Deposited: 11 Apr 2023 13:57 UTC
Last Modified: 12 Apr 2023 13:01 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/100820 (The current URI for this page, for reference purposes)
Moore, Simon J.: https://orcid.org/0000-0002-1968-206X
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