Directed Evolution of a Tryptophan 2,3-Dioxygenase for the Diastereoselective Monooxygenation of Tryptophans.
Angew Chem Int Ed Engl
; 59(8): 3043-3047, 2020 02 17.
Article
em En
| MEDLINE
| ID: mdl-31828916
ABSTRACT
Herein, we report an engineered enzyme that can monooxygenate unprotected tryptophan into the corresponding 3a-hydroxyhexahydropyrrolo[2,3-b]indole-2-carboxylic acid (HPIC) in a single, scalable step with excellent turnover number and diastereoselectivity. Taking advantage of directed evolution, we analyzed the stepwise oxygen-insertion mechanism of tryptophan 2,3-dioxygenases, and transformed tryptophan 2,3-dioxygenase from Xanthomonas campestris into a monooxygenase for oxidative cyclization of tryptophans. It was revealed that residue F51 is vital in determining the product ratio of HPIC to N'-formylkynurenine. Our reactions and purification procedures use no organic solvents, resulting in an eco-friendly method to prepare HPICs for further applications.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Triptofano
/
Triptofano Oxigenase
/
Oxigenases de Função Mista
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article