A Versatile Protein Scaffold Engineered for the Hierarchical Assembly of Robust and Highly Active Enzymes

Advanced Science, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 22, 2025

Abstract Scaffold proteins play immense roles in bringing enzymes together to enhance their properties. However, the direct fusion of scaffold with bulky guest may disrupt assembly process or diminish catalytic efficiency. Most self‐assembling protein scaffolds are engineered form structures beforehand, and then carry via different conjugation strategies vitro. Here, a robust is presented, from Methanococcus jannaschii using disulfide bonds, which efficiently assembles into higher‐order helices without additional chemistry bio‐conjugation When fused directly monomeric Endo‐1,4‐beta‐xylanase A, efficiency enzyme increased by 2.5 times enhanced thermostability. Additionally, integrating multimeric metalloenzyme nitrile hydratase overcame typical stability‐activity trade‐off such industrial enzyme, yielding three‐fold higher activity 28‐fold Structural analyses suggest that artificially made helical twist create new interface interactions provide concentration active sites enzymes. Further fluorescent pairs exhibited 12‐fold FRET efficiency, suggesting its potential for dual‐enzyme cascade applications. Overall, this study showcases simple yet powerful organizes hierarchical performance.

Language: Английский

A pivotal switch in β-domain determines the substrate selectivity of TPS genes involved in Gardenia jasminoides floral scent synthesis

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 288, P. 138333 - 138333

Published: Dec. 7, 2024

Language: Английский