A Plurizyme with Transaminase and Hydrolase Activity Catalyzes Cascade Reactions.
Angew Chem Int Ed Engl
; 61(37): e202207344, 2022 09 12.
Article
em En
| MEDLINE
| ID: mdl-35734849
ABSTRACT
Engineering dual-function single polypeptide catalysts with two abiotic or biotic catalytic entities (or combinations of both) supporting cascade reactions is becoming an important area of enzyme engineering and catalysis. Herein we present the development of a PluriZyme, TR2 E2 , with efficient native transaminase (kcat 69.49±1.77â
min-1 ) and artificial esterase (kcat 3908-0.41â
min-1 ) activities integrated into a single scaffold, and evaluate its utility in a cascade reaction. TR2 E2 (pHopt 8.0-9.5; Topt 60-65 °C) efficiently converts methyl 3-oxo-4-(2,4,5-trifluorophenyl)butanoate into 3-(R)-amino-4-(2,4,5-trifluorophenyl)butanoic acid, a crucial intermediate for the synthesis of antidiabetic drugs. The reaction proceeds through the conversion of the ß-keto ester into the ß-keto acid at the hydrolytic site and subsequently into the ß-amino acid (e.e. >99 %) at the transaminase site. The catalytic power of the TR2 E2 PluriZyme was proven with a set of ß-keto esters, demonstrating the potential of such designs to address bioinspired cascade reactions.
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Base de dados:
MEDLINE
Assunto principal:
Aminoácidos
/
Transaminases
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article