Structure of the Reductase Domain of a fungal Carboxylic Acid Reductase and its Substrate Scope in Thioester and Aldehyde Reduction

Bastian Daniel, Chiam Hashem, Marlene Leithold, Theo Sagmeister, Adrian Tripp, Holly Stolterfoht-Stock, Julia Messenlehner, Ronan Keegan, Christoph K. Winkler, Jonathan Guyang Ling, Sabry H.H. Younes, Gustav Oberdorfer, Farah Diba Abu Bakar, Karl Gruber, Tea Pavkov-Keller*, Margit Winkler*

*Korrespondierende/r Autor/-in für diese Arbeit

Publikation: Beitrag in einer FachzeitschriftArtikelBegutachtung

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 Neurospora crassa (Nc). The NcCAR R-domain revealed activity for N-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 NcCAR R-domain reveals a tunnel that putatively harbors the phosphopantetheinylacyl-intermediate, which is in good agreement with docking experiments performed with the minimal substrate. In vitro 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 NcCAR reduced aldehydes to primary alcohols. In conclusion, aldehyde overreduction can no longer be attributed exclusively to the host background
Originalspracheenglisch
Seiten (von - bis)15668–15674
Seitenumfang7
FachzeitschriftACS Catalysis
Jahrgang12
Ausgabenummer24
DOIs
PublikationsstatusVeröffentlicht - 16 Dez. 2022

ASJC Scopus subject areas

  • Chemie (insg.)
  • Katalyse

Fields of Expertise

  • Human- & Biotechnology

Kooperationen

  • BioTechMed-Graz

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