8AEP

Reductase domain of the carboxylate reductase of Neurospora crassa

Summary for 8AEP

• Entry DOI: 10.2210/pdb8aep/pdb
• Descriptor: Acetyl-CoA synthetase-like protein, CHLORIDE ION, SULFATE ION, ... (4 entities in total)
• Functional Keywords: oxidereductase, carboxylate reductase, oxidoreductase
• Biological source: Neurospora crassa
• Total number of polymer chains: 2
• Total formula weight: 90068.03

Authors

Daniel, B.,Schrufer, A.,Marlene, L.,Sagmeister, T.,Pavkov-Keller, T. (deposition date: 2022-07-13, release date: 2023-03-08, Last modification date: 2024-02-07)

Primary citation

Daniel, B.,Hashem, C.,Leithold, M.,Sagmeister, T.,Tripp, A.,Stolterfoht-Stock, H.,Messenlehner, J.,Keegan, R.,Winkler, C.K.,Ling, J.G.,Younes, S.H.H.,Oberdorfer, G.,Abu Bakar, F.D.,Gruber, K.,Pavkov-Keller, T.,Winkler, M.
Structure of the Reductase Domain of a Fungal Carboxylic Acid Reductase and Its Substrate Scope in Thioester and Aldehyde Reduction.
Acs Catalysis, 12:15668-15674, 2022

PubMed Abstract: The synthesis of aldehydes from carboxylic acids has long been a challenge in chemistry. In contrast to the harsh chemically driven reduction, enzymes such as carboxylic acid reductases (CARs) are considered appealing biocatalysts for aldehyde production. Although structures of single- and didomains of microbial CARs have been reported, to date no full-length protein structure has been elucidated. In this study, we aimed to obtain structural and functional information regarding the reductase (R) domain of a CAR from the fungus (). The CAR R-domain revealed activity for -acetylcysteamine thioester (S-(2-acetamidoethyl) benzothioate), which mimics the phosphopantetheinylacyl-intermediate and can be anticipated as the minimal substrate for thioester reduction by CARs. The determined crystal structure of the CAR R-domain reveals a tunnel that putatively harbors the phosphopantetheinylacyl-intermediate, which is in good agreement with docking experiments performed with the minimal substrate. studies were performed with this highly purified R-domain and NADPH, demonstrating carbonyl reduction activity. The R-domain was able to accept not only a simple aromatic ketone but also benzaldehyde and octanal, which are typically considered to be the final product of carboxylic acid reduction by CAR. Also, the full-length CAR reduced aldehydes to primary alcohols. In conclusion, aldehyde overreduction can no longer be attributed exclusively to the host background.

PubMed: 37180375
DOI: 10.1021/acscatal.2c04426

Experimental method

X-RAY DIFFRACTION (2.3 Å)