8OT8

Alditol oxidase from Actinomycetota bacterium in complex with D-xylulose

8OT8 の概要

• エントリーDOI: 10.2210/pdb8ot8/pdb
• 分子名称: FAD-binding protein, FLAVIN-ADENINE DINUCLEOTIDE, D-XYLULOSE, ... (4 entities in total)
• 機能のキーワード: alditol oxidase, flavoprotein, xylulose
• 由来する生物種: Actinomycetota bacterium
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 188447.06

構造登録者

Rotilio, L.,Mattevi, A. (登録日: 2023-04-20, 公開日: 2024-02-28)

主引用文献

Santema, L.L.,Rotilio, L.,Xiang, R.,Tjallinks, G.,Guallar, V.,Mattevi, A.,Fraaije, M.W.
Discovery and biochemical characterization of thermostable glycerol oxidases.
Appl.Microbiol.Biotechnol., 108:61-61, 2024

PubMed Abstract: Alditol oxidases are promising tools for the biocatalytic oxidation of glycerol to more valuable chemicals. By integrating in silico bioprospecting with cell-free protein synthesis and activity screening, an effective pipeline was developed to rapidly identify enzymes that are active on glycerol. Three thermostable alditol oxidases from Actinobacteria Bacterium, Streptomyces thermoviolaceus, and Thermostaphylospora chromogena active on glycerol were discovered. The characterization of these three flavoenzymes demonstrated their glycerol oxidation activities, preference for alkaline conditions, and excellent thermostabilities with melting temperatures higher than 75 °C. Structural elucidation of the alditol oxidase from Actinobacteria Bacterium highlighted a constellation of side chains that engage the substrate through several hydrogen bonds, a histidine residue covalently bound to the FAD prosthetic group, and a tunnel leading to the active site. Upon computational simulations of substrate binding, a double mutant targeting a residue pair at the tunnel entrance was created and found to display an improved thermal stability and catalytic efficiency for glycerol oxidation. The hereby described alditol oxidases form a valuable panel of oxidative biocatalysts that can perform regioselective oxidation of glycerol and other polyols. KEY POINTS: • Rapid pipeline designed to identify putative oxidases • Biochemical and structural characterization of alditol oxidases • Glycerol oxidation to more valuable derivatives.

PubMed: 38183484
DOI: 10.1007/s00253-023-12883-9

実験手法

X-RAY DIFFRACTION (2.4 Å)