Designing a novel (R)-ω-transaminase for asymmetric synthesis of sitagliptin intermediate via motif swapping and semi-rational design.
(R)-ω-transaminase
Enantioselectivity
Motif swapping
Protein engineering
Sitagliptin
Journal
International journal of biological macromolecules
ISSN: 1879-0003
Titre abrégé: Int J Biol Macromol
Pays: Netherlands
ID NLM: 7909578
Informations de publication
Date de publication:
31 Dec 2023
31 Dec 2023
Historique:
received:
16
06
2023
revised:
25
09
2023
accepted:
09
10
2023
medline:
23
11
2023
pubmed:
12
10
2023
entrez:
11
10
2023
Statut:
ppublish
Résumé
The application of (R)-ω-transaminases as biocatalysts for chiral amine synthesis has been hampered by inadequate stereoselectivity and narrow substrate spectrum. Herein, an effective evolution strategy for (R)-ω-transaminase designing for the asymmetric synthesis of sitagliptin intermediate is presented. Since natural transaminases lack activity toward bulky prositagliptin ketone, transaminase scaffolds with catalytic machinery and activity toward the truncated prositagliptin ketone were firstly screened based on substrate walking principle. A transaminase chimera was established synchronously conferring catalytic activity and (R)-selectivity toward prositagliptin ketone through motif swapping, followed by stepwise evolution. The process resulted in a "best" engineered variant MwTA
Identifiants
pubmed: 37820904
pii: S0141-8130(23)04245-9
doi: 10.1016/j.ijbiomac.2023.127348
pii:
doi:
Substances chimiques
Transaminases
EC 2.6.1.-
Sitagliptin Phosphate
TS63EW8X6F
Ketones
0
Amines
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
127348Informations de copyright
Copyright © 2023 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this work.