5YBQ

Fe(II)/(alpha)ketoglutarate-dependent dioxygenase PrhA-V150L/A232S in complex with preaustinoid A2

Summary for 5YBQ

• Entry DOI: 10.2210/pdb5ybq/pdb
• Descriptor: PrhA, FE (III) ION, 2-OXOGLUTARIC ACID, ... (5 entities in total)
• Functional Keywords: alpha-kegoglutarate-dependent dioxygenase, oxidoreductase
• Biological source: Penicillium brasilianum
• Total number of polymer chains: 2
• Total formula weight: 71714.59

Authors

Nakashima, Y.,Senda, M. (deposition date: 2017-09-05, release date: 2018-01-24, Last modification date: 2023-11-22)

Primary citation

Nakashima, Y.,Mori, T.,Nakamura, H.,Awakawa, T.,Hoshino, S.,Senda, M.,Senda, T.,Abe, I.
Structure function and engineering of multifunctional non-heme iron dependent oxygenases in fungal meroterpenoid biosynthesis.
Nat Commun, 9:104-104, 2018

PubMed Abstract: Non-heme iron and α-ketoglutarate (αKG) oxygenases catalyze remarkably diverse reactions using a single ferrous ion cofactor. A major challenge in studying this versatile family of enzymes is to understand their structure-function relationship. AusE from Aspergillus nidulans and PrhA from Penicillium brasilianum are two highly homologous Fe(II)/αKG oxygenases in fungal meroterpenoid biosynthetic pathways that use preaustinoid A1 as a common substrate to catalyze divergent rearrangement reactions to form the spiro-lactone in austinol and cycloheptadiene moiety in paraherquonin, respectively. Herein, we report the comparative structural study of AusE and PrhA, which led to the identification of three key active site residues that control their reactivity. Structure-guided mutagenesis of these residues results in successful interconversion of AusE and PrhA functions as well as generation of the PrhA double and triple mutants with expanded catalytic repertoire. Manipulation of the multifunctional Fe(II)/αKG oxygenases thus provides an excellent platform for the future development of biocatalysts.

PubMed: 29317628
DOI: 10.1038/s41467-017-02371-w

Experimental method

X-RAY DIFFRACTION (2.25 Å)