| RRC ID |
66220
|
| 著者 |
Bernhardsgrütter I, Schell K, Peter DM, Borjian F, Saez DA, Vöhringer-Martinez E, Erb TJ.
|
| タイトル |
Awakening the Sleeping Carboxylase Function of Enzymes: Engineering the Natural CO2-Binding Potential of Reductases.
|
| ジャーナル |
J Am Chem Soc
|
| 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.
|
| 巻・号 |
141(25)
|
| ページ |
9778-9782
|
| 公開日 |
2019-6-26
|
| DOI |
10.1021/jacs.9b03431
|
| PMID |
31188584
|
| PMC |
PMC6650136
|
| MeSH |
Archaea / enzymology
Archaeal Proteins / chemistry
Archaeal Proteins / genetics
Archaeal Proteins / metabolism
Bacterial Proteins / chemistry
Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Carbon Cycle
Carbon Dioxide / chemistry
Carbon Dioxide / metabolism
Carboxy-Lyases / chemistry*
Carboxy-Lyases / genetics
Carboxy-Lyases / metabolism
Catalytic Domain / genetics
Kinetics
Molecular Dynamics Simulation
Mutagenesis, Site-Directed
Oxidoreductases / chemistry*
Oxidoreductases / genetics
Oxidoreductases / metabolism
Sphingomonadaceae / enzymology
Water / chemistry
Water / metabolism
|
| IF |
14.612
|
| リソース情報 |
| 原核生物(大腸菌) |
|
