Loop engineering of enzymes to control their immobilization and ultimately fabricate more efficient heterogeneous biocatalysts.

ABSTRACT: Enzyme immobilization is indispensable for enhancing enzyme performance in various industrial applications. Typically, enzymes require specific spatial arrangements for optimal functionality, underscoring the importance of correct orientation. Despite well-known N- or C-terminus tailoring techniques, alternatives for achieving orientation control are limited. Here, we propose a novel approach that tailors the enzyme surface with engineered His-rich loops. To that aim, we first solve the X-ray crystal structure of a hexameric alcohol dehydrogenase from Thermus thermophilus HB27 (TtHBDH) (PDB: 9FBD). Guided by this 3D structure, we engineer the enzyme surface with a new loop enriched with six His residues to control enzyme orientation. Molecular dynamics simulations reveal that the engineere

SUBMITTER: Zeballos N 

PROVIDER: S-EPMC11751856 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature