Insight into the broadened substrate scope of nitrile hydratase by static and dynamic structure analysis.
Journal
Chemical science
ISSN: 2041-6520
Titre abrégé: Chem Sci
Pays: England
ID NLM: 101545951
Informations de publication
Date de publication:
20 Jul 2022
20 Jul 2022
Historique:
received:
25
04
2022
accepted:
28
06
2022
entrez:
3
8
2022
pubmed:
4
8
2022
medline:
4
8
2022
Statut:
epublish
Résumé
The narrow substrate scope limits the wide industrial application of enzymes. Here, we successfully broadened the substrate scope of a nitrile hydratase (NHase) through mutation of two tunnel entrance residues based on rational tunnel calculation. Two variants, with increased specific activity, especially toward bulky substrates, were obtained. Crystal structure analysis revealed that the mutations led to the expansion of the tunnel entrance, which might be conducive to substrate entry. More importantly, molecular dynamics simulations illustrated that the mutations introduced anti-correlated movements to the regions around the substrate tunnel and the active site, which would promote substrate access during the dynamic process of catalysis. Additionally, mutations on the corresponding tunnel entrance residues on other NHases also enhanced their activity toward bulky substrates. These results not only revealed that residues located at the enzyme surface were a key factor in enzyme catalytic performance, but also provided dynamic evidence for insight into enzyme substrate scope broadening.
Identifiants
pubmed: 35919716
doi: 10.1039/d2sc02319a
pii: d2sc02319a
pmc: PMC9297474
doi:
Types de publication
Journal Article
Langues
eng
Pagination
8417-8428Informations de copyright
This journal is © The Royal Society of Chemistry.
Déclaration de conflit d'intérêts
The authors declare no competing interests.
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