Substitutability of metal-binding sites in an alcohol dehydrogenase / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1518-1526, 2022.
Artículo
en Chino
| WPRIM
| ID: wpr-927797
ABSTRACT
Covalently anchoring of a ligand/metal via polar amino acid side chain(s) is often observed in metalloenzyme, while the substitutability of metal-binding sites remains elusive. In this study, we utilized a zinc-dependent alcohol dehydrogenase from Thermoanaerobacter brockii (TbSADH) as a model enzyme, analyzed the sequence conservation of the three residues Cys37, His59, and Asp150 that bind the zinc ion, and constructed the mutant library. After experimental validation, three out of 224 clones, which showed comparative conversion and ee values as the wild-type enzyme in the asymmetric reduction of the model substrate tetrahydrofuran-3-one, were screened out. The results reveal that the metal-binding sites in TbSADH are substitutable without tradeoff in activity and stereoselectivity, which lay a foundation for designing ADH-catalyzed new reactions via metal ion replacement.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Zinc
/
Alcohol Deshidrogenasa
/
Dominio Catalítico
/
Dominios Proteicos
/
Ligandos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2022
Tipo del documento:
Artículo
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