9H92

FAD-dependent oxidase sorD

Summary for 9H92

• Entry DOI: 10.2210/pdb9h92/pdb
• Descriptor: FAD-linked oxidoreductase sorD, alpha-D-mannopyranose-(1-6)-alpha-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, ... (7 entities in total)
• Functional Keywords: flavin, fad, natural product, oxidase, sorbicillinoids, oxidoreductase
• Biological source: Penicillium rubens Wisconsin 54-1255
• Total number of polymer chains: 1
• Total formula weight: 53328.09

Authors

Tjallinks, G.,Mattevi, A. (deposition date: 2024-10-29, release date: 2025-03-19, Last modification date: 2025-04-02)

Primary citation

Tjallinks, G.,Angeleri, N.,Nguyen, Q.T.,Mannucci, B.,Arentshorst, M.,Visser, J.,Ram, A.F.J.,Fraaije, M.W.,Mattevi, A.
Structural and Mechanistic Characterization of the Flavin-Dependent Monooxygenase and Oxidase Involved in Sorbicillinoid Biosynthesis.
Acs Chem.Biol., 20:646-655, 2025

PubMed Abstract: Sorbicillinoids are yellow secondary metabolites synthesized through an elegant combination of enzymatic and spontaneous biochemical processes. The flavin-dependent monooxygenase SorC and oxidase SorD are crucial in this interplay, enabling the generation of a diverse array of functionally complex sorbicillinoids. By solving the crystal structures of SorC and SorD from with sorbicillin bound in the active site, we describe the catalytically active binding conformations, crucial for attaining enantioselective and stereoselective control in these enzymatic reactions. The structure of SorC was resolved with the cofactor FAD in its state, which allowed us to identify key residues that modulate flavin mobility and other conformational changes. Catalytic residues of SorC were also confirmed by detailed characterization of wild-type and several SorC variants. Meanwhile, using a CRISPR/Cas9-based multicopy-genome integration system, we could heterologously express the flavin-dependent oxidase SorD from in with high yields and purity. This allowed us to obtain the crystal structure of SorD with sorbicillin bound in a viable catalytic conformation. Structural analysis of the obtained complex provided insights into the substrate binding pose and highlighted potentially critical active site residues. Ultimately, having both SorC and SorD at our disposal enabled us to investigate their functions and interplays in the biosynthesis of a vast array of functionally complex sorbicillinoids.

PubMed: 40052414
DOI: 10.1021/acschembio.4c00783

Experimental method

X-RAY DIFFRACTION (1.55 Å)