8S1J

Crystal structure of t-anethole oxygenase from Stenotrophomonas maltophilia

8S1J の概要

• エントリーDOI: 10.2210/pdb8s1j/pdb
• 分子名称: Trans-anethole oxygenase, PROTOPORPHYRIN IX CONTAINING FE, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: oxygenase, c=c cleavage, bacterial, heme, oxidoreductase
• 由来する生物種: Stenotrophomonas maltophilia
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 72077.79

構造登録者

Rozeboom, H.J.,Fraaije, M.W. (登録日: 2024-02-15, 公開日: 2024-08-14)

主引用文献

Purwani, N.N.,Rozeboom, H.J.,Willers, V.P.,Wijma, H.J.,Fraaije, M.W.
Discovery of a new class of bacterial heme-containing CC cleaving oxygenases.
N Biotechnol, 83:82-90, 2024

PubMed Abstract: Previously, some bacteria were shown to harbour enzymes capable of catalysing the oxidative cleavage of the double bond of t-anethole and related compounds. The cofactor dependence of these enzymes remained enigmatic due to a lack of biochemical information. We report on catalytic and structural details of a representative of this group of oxidative enzymes: t-anethole oxygenase from Stenotrophomonas maltophilia (TAO). The bacterial enzyme could be recombinantly expressed and purified, enabling a detailed biochemical study that has settled the dispute on its cofactor dependence. We have established that TAO contains a tightly bound b-type heme and merely depends on dioxygen for catalysis. It was found to accept t-anethole, isoeugenol and O-methyl isoeugenol as substrates, all being converted into the corresponding aromatic aldehydes without the need of any cofactor regeneration. The elucidated crystal structure of TAO has revealed that it contains a unique active site architecture that is conserved for this distinct class of heme-containing bacterial oxygenases. Similar to other hemoproteins, TAO has a histidine (His121) as proximal ligand. Yet, unique for TAOs, an arginine (Arg89) is located at the distal axial position. Site directed mutagenesis confirmed crucial roles for these heme-liganding residues and other residues that form the substrate binding pocket. In conclusion, the results reported here reveal a new class of bacterial heme-containing oxygenases that can be used for the cleavage of alkene double bonds, analogous to ozonolysis in organic chemistry.

PubMed: 39053683
DOI: 10.1016/j.nbt.2024.07.002

実験手法

X-RAY DIFFRACTION (1.83 Å)