Awakening the Sleeping Carboxylase Function of Enzymes: Engineering the Natural CO2-Binding Potential of Reductases

Bernhardsgrütter I.; Schell K.; Peter D.M.; Borjian F.; Saez D.A.; Vöhringer-Martinez E.; Erb T.J.

Abstract

Developing new carbon dioxide (CO2) fixing enzymes is a prerequisite to create new biocatalysts for diverse applications in chemistry, biotechnology and synthetic biology. Here we used bioinformatics to identify a "sleeping carboxylase function" in the superfamily of medium-chain dehydrogenases/reductases (MDR), i.e. enzymes that possess a low carboxylation side activity next to their original enzyme reaction. We show that propionyl-CoA synthase from Erythrobacter sp. NAP1, as well as an acrylyl-CoA reductase from Nitrosopumilus maritimus possess carboxylation yields of 3 +/- 1 and 4.5 +/- 0.9%. We use rational design to engineer these enzymes further into carboxylases by increasing interactions of the proteins with CO2 and suppressing diffusion of water to the active site. The engineered carboxylases show improved CO2-binding and kinetic parameters comparable to naturally existing CO2-fixing enzymes. Our results provide a strategy to develop novel CO2-fixing enzymes and shed light on the emergence of natural carboxylases during evolution.

Más información

Título según WOS: Awakening the Sleeping Carboxylase Function of Enzymes: Engineering the Natural CO2-Binding Potential of Reductases
Título según SCOPUS: Awakening the Sleeping Carboxylase Function of Enzymes: Engineering the Natural CO2-Binding Potential of Reductases
Título de la Revista: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volumen: 141
Número: 25
Editorial: AMER CHEMICAL SOC
Fecha de publicación: 2019
Página de inicio: 9778
Página final: 9782
Idioma: English
DOI:

10.1021/jacs.9b03431

Notas: ISI, SCOPUS