2026-06-08T12:56:11Zhttps://docta.ucm.es/rest/oai/requestoai:docta.ucm.es:20.500.14352/1054532024-07-03T00:07:32Zcom_20.500.14352_14col_20.500.14352_15
Docta Complutense
author
Ledesma Fernández, Alba
author
Velasco Lozano, Susana
author
Campos Muelas, Pedro
author
Madrid González, Ricardo
author
López Gallego, Fernando
author
Cortajarena, Aitziber L.
2024-07-02T15:02:41Z
2024-07-02T15:02:41Z
2024-04-12
Ledesma-Fernandez A,Velasco-Lozano S, Campos-Muelas P, Madrid R,L opez-Gallego F, Cortajarena AL. Engineering bio-brick protein scaffolds for organizing enzymeassemblies. Protein Science. 2024;33(5):e4984.
0961-83681469-896X
10.1002/pro.4984
https://hdl.handle.net/20.500.14352/105453
https://doi.org/10.1002/pro.4984
https://pubmed.ncbi.nlm.nih.gov/38607190/
Enzyme scaffolding is an emerging approach for enhancing the catalytic efficiency of multi‐enzymatic cascades by controlling their spatial organization and stoichiometry. This study introduces a novel family of engineered SCAffolding Bricks, named SCABs, utilizing the consensus tetratricopeptide repeat (CTPR) domain for organized multi‐enzyme systems. Two SCAB systems are developed, one employing head‐to‐tail interactions with reversible covalent disulfide bonds, the other relying on non‐covalent metal‐driven assembly via engineered metal coordinating interfaces. Enzymes are directly fused to SCAB modules, triggering assembly in a non‐reducing environment or by metal presence. A proof‐of‐concept with formate dehydrogenase (FDH) and L‐alanine dehydrogenase (AlaDH) shows enhanced specific productivity by 3.6‐fold compared to free enzymes, with the covalent stapling outperforming the metal‐driven assembly. This enhancement likely stems from higher‐order supramolecular assembly and improved NADH cofactor regeneration, resulting in more efficient cascades. This study underscores the potential of protein engineering to tailor scaffolds, leveraging supramolecular spatial‐organizing tools, for more efficient enzymatic cascade reactions.
eng
Engineering bio‐brick protein scaffolds for organizing enzyme assemblies
journal article
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URL
https://docta.ucm.es/bitstreams/9346206c-c338-428e-9b26-0ea976aea890/download
File
MD5
a0abb686ad54f5a210e1fb3830a800a4
13930961
application/pdf
Engineering_bio-brick_protein.pdf