Structure and Electron Transfer of Quinoproteins and application to Bio-electrode Devices

醌蛋白的结构和电子转移及其在生物电极器件中的应用

• 批准号: 15380082
• 负责人: KANO Kenji
• 金额: $ 8.64万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380082/
• 关键词: Histamine dehydrogenase; 6-S-cyteinyl-FMN; Expression system; Quinohemoproteins; Amine dehydrogenase; Alcohol dehydrogenase; Biosensors; Biofuel cells; 直接電子移動型酵素電極反応

1)Structural analysis and Electron Transfer Characteristics of Histamine Dehydrogenase (HmDH)It has been revealed that HmDH has 6-S-cyteinyl-FMN and 4Fe-4S cluster as redox prosthetic groups. Under neutral conditions, 2-electron reductions occurs per monomer by the substrate, while the number of the electron is switched to unit over pH 9.5. The properties can be explained in terms of the change in the redox potentials. The kinetics of half-reduction has been analyzed with stopped flow method. The holo form of HmDH has been expressed in E.coli in the purity of 100%. Site-directed mutation improve the specificity of the enzyme.2)Electron Transfer of QuinohemoproteinsPositive charge in the vicinity of the active site of quinohemoprotein amine dehydrogenase (QH-AmDH) plays an important role in the electron transfer from to electron acceptor. The direct electron transfer from QH-AmDH to electrodes was not observed. The redox potentials of quinohemoproteins alcohol dehydrogenase depend on pH. This would be due to the structural change of the enzyme.3)Application to Biosensor and Biofuel CellsIt has been found that mutant HmDH works as a good catalysis of mediated biosensor for histamine with high specificity. Selectivity of mediator reduced the interference. The dehydrogenases invested here were utilized also as catalysts in bioanode of biofuel cells, where bilirubin oxidase (BOD) was used as cathode enzyme. Direct electron transfer of BOD is affected by the electrode materials. The current density were increased up few ten mA cm^<-2>.

1)组胺脱氢酶（HmDH）的结构分析和电子转移特性研究表明，HmDH含有6-S-半胱氨酰-FMN和4Fe-4S簇作为氧化还原辅基。在中性条件下，每个单体通过底物发生2-电子还原，而电子数在pH 9.5以上切换为单位。这些性质可以用氧化还原电位的变化来解释。用停流法分析了半还原动力学。在大肠杆菌中表达了HmDH的完整形式，纯度为100%。定点突变提高了酶的特异性。2）奎诺血蛋白的电子转移奎诺血蛋白胺脱氢酶（QH-AmDH）活性位点附近的正电荷在电子从受体到受体的转移中起着重要作用。没有观察到从QH-AmDH到电极的直接电子转移。醌血红素蛋白醇脱氢酶的氧化还原电位依赖于pH值，这可能是由于酶的结构发生了变化。3）生物传感器和生物燃料电池的应用研究发现，突变型HmDH作为介导型组胺生物传感器的催化剂，具有很好的特异性。介质的选择性降低了干扰。在生物燃料电池的生物阳极中，胆红素氧化酶（BOD）被用作阴极酶。电极材料对BOD的直接电子转移有影响。电流密度增加了几十mA cm ↑ [2]<-2>。

项目成果

Kenji Kano: "Redox Properties of Quinohemoprotein Amine Dehydrogenase from Paracoccus denitrifficans"Biochim.Biophys.Acta. 1647・1/2. 289-296 (2003)

Kenji Kano：“来自脱氮副球菌的醌血蛋白胺脱氢酶的氧化还原特性”Biochim.Biophys.Acta 1647·1/2（2003）。

Mediated spectroelectrochemical titration of proteins for redox potential measurements by a separator-less one-compartment bulk electrolysis method.

• DOI: 10.1016/j.ab.2004.11.017
• 发表时间: 2005-02
• 期刊: Analytical biochemistry
• 影响因子: 2.9
• 作者: S. Tsujimura;Atsushi Kuriyama;N. Fujieda;K. Kano;T. Ikeda

Separator-less One-compartment Bulk Electrolysis with a Small Auxiliary Electrode and Its Application to Spectroelectrochemistry

小辅助电极无隔板单室本体电解及其在光谱电化学中的应用

• 发表时间: 2004
• 期刊: Electrochemistry 72(7)
• 作者: Y.Kamitaka;S.Tsujimura;T.Ikeda;K.Kano;Yong-Fu Wang;Seiya Tsujimura;Seiya Tsujimura;Seiya Tsujimura;Seiya Tsujimura;Shinki Kojima;Seiya Tsujimura;Atsushi Kuriyama
• 通讯作者: Atsushi Kuriyama

Kinetic Study of Direct Bioelectrocatalysis of Dioxygen Reduction with Bilirubin Oxidase at Carbon Electrodes

• DOI: 10.5796/electrochemistry.72.437
• 发表时间: 2004-06
• 期刊: Electrochemistry
• 影响因子: 2.5
• 作者: S. Tsujimura;T. Nakagawa;K. Kano;T. Ikeda

Current Status of Batteries in Next Generation

下一代电池的现状

• 发表时间: 2005
• 期刊: E.T.S.
• 作者: Hayase;K.;Tujioka;K.;Yokogoshi;H.;Sayaka Nanjo et al.;横越英彦;横越英彦;Seiya Tsujimura et al.;横越英彦;横越英彦 編集・分担執筆;Seiya Tsujimura et al.
• 通讯作者: Seiya Tsujimura et al.