6H3O

Alcohol oxidase from Phanerochaete chrysosporium mutant F101S

6H3O の概要

• エントリーDOI: 10.2210/pdb6h3o/pdb
• 分子名称: Alcohol oxidase, FLAVIN-ADENINE DINUCLEOTIDE, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: alcohol oxidase, octamer, fad-binding domain, mutant f101s, oxidoreductase
• 由来する生物種: Phanerochaete chrysosporium
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 587051.03

構造登録者

Nguyen, Q.-T.,Romero, E.,Dijkman, W.P.,de Vasconcellos, S.P.,Binda, C.,Mattevi, A.,Fraaije, M.W. (登録日: 2018-07-19, 公開日: 2018-10-10, 最終更新日: 2024-01-17)

主引用文献

Nguyen, Q.T.,Romero, E.,Dijkman, W.P.,de Vasconcellos, S.P.,Binda, C.,Mattevi, A.,Fraaije, M.W.
Structure-Based Engineering of Phanerochaete chrysosporium Alcohol Oxidase for Enhanced Oxidative Power toward Glycerol.
Biochemistry, 57:6209-6218, 2018

PubMed Abstract: Glycerol is a major byproduct of biodiesel production, and enzymes that oxidize this compound have been long sought after. The recently described alcohol oxidase from the white-rot basidiomycete Phanerochaete chrysosporium (PcAOX) was reported to feature very mild activity on glycerol. Here, we describe the comprehensive structural and biochemical characterization of this enzyme. PcAOX was expressed in Escherichia coli in high yields and displayed high thermostability. Steady-state kinetics revealed that PcAOX is highly active toward methanol, ethanol, and 1-propanol ( k = 18, 19, and 11 s, respectively), but showed very limited activity toward glycerol ( k = 0.2 s at 2 M substrate). The crystal structure of the homo-octameric PcAOX was determined at a resolution of 2.6 Å. The catalytic center is a remarkable solvent-inaccessible cavity located at the re side of the flavin cofactor. Its small size explains the observed preference for methanol and ethanol as best substrates. These findings led us to design several cavity-enlarging mutants with significantly improved activity toward glycerol. Among them, the F101S variant had a high k value of 3 s, retaining a high degree of thermostability. The crystal structure of F101S PcAOX was solved, confirming the site of mutation and the larger substrate-binding pocket. Our data demonstrate that PcAOX is a very promising enzyme for glycerol biotransformation.

PubMed: 30272958
DOI: 10.1021/acs.biochem.8b00918

実験手法

X-RAY DIFFRACTION (2.5 Å)