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Updated: May 4, 2026
![Protein Film Infrared Electrochemistry Demonstrated for Study of H2 Oxidation by a [NiFe] Hydrogenase](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F55858.jpg&w=3840&q=50)
Protein Film Infrared Electrochemistry Demonstrated for Study of H2 Oxidation by a [NiFe] Hydrogenase
Published on: December 4, 2017
H2O2-dependent substrate oxidation by an engineered diiron site in a bacterial hemerythrin
Yasunori Okamoto1, Akira Onoda, Hiroshi Sugimoto
1Department of Applied Chemistry, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan. onoda@chem.eng.osaka-u.ac.jp thayashi@chem.eng.osaka-u.ac.jp.
Researchers engineered a diiron site in DcrH-Hr to oxidize external substrates using hydrogen peroxide (H2O2). The modified enzyme successfully oxidized guaiacol and 1,4-cyclohexadiene, demonstrating a new catalytic capability.
Area of Science:
- Biochemistry
- Enzyme Engineering
- Oxidation Reactions
Background:
- The carboxylate-bridged diiron site in DcrH-Hr is crucial for O2 binding.
- Engineering efforts aim to expand the catalytic functions of metalloenzymes.
Purpose of the Study:
- To engineer the DcrH-Hr diiron site for H2O2-dependent oxidation of external substrates.
- To investigate the impact of introducing a histidine residue on enzyme activity.
Main Methods:
- Site-directed mutagenesis was used to replace Ile119 with Histidine (His).
- The engineered I119H variant was assayed for oxidation of guaiacol and 1,4-cyclohexadiene in the presence of H2O2.
Main Results:
- The I119H variant of DcrH-Hr exhibited H2O2-dependent oxidation activity.
- Specific substrates, guaiacol and 1,4-cyclohexadiene, were successfully oxidized by the engineered enzyme.
Conclusions:
- Engineering the diiron site in DcrH-Hr can confer new substrate oxidation capabilities.
- The introduction of a histidine residue near the diiron site is key to this modified function.
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