Development of Novel Artificial Dinuclear Metalloenzymes and Application to Catalytic Oxidation Reactions

• Project/Area Number: 25620044
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Inorganic chemistry
• Research Institution: Osaka University
• Principal Investigator: ITOH Shinobu 大阪大学, 工学(系)研究科(研究院), 教授 (30184659)
• Co-Investigator(Renkei-kenkyūsha): SUGIMOTO Hideki 大阪大学, 大学院工学研究科, 准教授 (00315970) ; FUJIEDA Nobutaka 大阪大学, 大学院工学研究科, 助教 (00452318)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: 人工金属酵素 / 酸化酵素 / 遷移金属錯体 / 金属結合タンパク質 / チロシナーゼ / 超好熱菌 / コウジ菌 / 二核金属酵素 / 酸化反応 / 触媒反応 / 超好熱菌由来タンパク質

Outline of Final Research Achievements

In this study, novel artificial metalloenzymes have been developed using dicopper copper monooxygenase tyrosinase and metal-binding protein TM1459 from hyperthermophiles. The crystal structures of the deoxy-, met-, and oxy-forms of tyrosinase from Aspergillus oryzae and its mutants have been determined, and their spectroscopic and magnetic characteristics as well as the catalytic activity have been explored in detail. On the basis of those information, artificial dicopper enzymes have been developed for the catalytic phenol oxidation reactions. Furthermore, novel metalloenzymes containing several transition-metal ions such as Zn, Cu, Ni, Co, Fe, Mn, Cr, V, Ru, Os have been developed by using a simple metal-binding protein from Thermotoga maritima, and their crustal structures as well as the spectroscopic, magnetic properties and catalytic activity have been examined in detail.

Research Products

• [Journal Article] Redox behavior of novel nickel and palladium complexes supported by trianionic non-innocent ligand containing β-diketiminate and phenol groups (2015)
  • Author(s): Shinobu Itoh, et al.
  • Journal Title: J. Porphyrins Phthalocyanines Volume: 19 Issue: 01-03 Pages: 377-387
  • DOI: 10.1142/s1088424615500248
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Electrochemical reduction of dioxygen by copper complexes with pyridylalkylamine ligands dissolved in aqueous buffer solution: relationship between activity and redox potential (2014)
  • Author(s): Shinobu Itoh, et al.
  • Journal Title: Dalton Trans. Volume: 43 Issue: 28 Pages: 10705-10709
  • DOI: 10.1039/c4dt00606b
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Geometric control of nuclearity in copper(I)/dioxygen chemistry (2014)
  • Author(s): T. Abe, Y. Morimoto, T. Tano, K. Mieda, H. Sugimotok N. Fujieda, T. Ogura, and S. Itoh
  • Journal Title: Inorganic Chemistry Volume: 53 Issue: 16 Pages: 8786-8794
  • DOI: 10.1021/ic501461n
  • Peer Reviewed
• [Journal Article] Copper complexes supported by an N2S-tridentate ligand as active site models of copper monooxygenases (2014)
  • Author(s): Shinobu Itoh, et al.
  • Journal Title: Dalton Trans. Volume: 42 Issue: 12 Pages: 4871-4877
  • DOI: 10.1039/c3dt52952e
  • Peer Reviewed
• [Journal Article] Activation Mechanism of MelB Tyrosinase from Aspergillus oryzae by acidic treatment (2013)
  • Author(s): Fujieda, N., Murata, M., Yabuta, S., Ikeda, T., Shimokawa, C., Nakamura, Y., Hata, Y., and Itoh, S.
  • Journal Title: J. Biol. Inorg. Chem. Volume: 18 Issue: 1 Pages: 19-26
  • DOI: 10.1007/s00775-012-0945-5
  • Peer Reviewed
• [Journal Article] Crystal Structures Of Copper-Depleted And Copper-Bound Fungal Pro-Tyrosinase: Insights Into Endogenous Cysteine-Dependent Copper Incorporation (2013)
  • Author(s): Shinobu Itoh, et al.
  • Journal Title: J. Biol.Chem. Volume: 288 Issue: 30 Pages: 22128-22140
  • DOI: 10.1074/jbc.m113.477612
  • Peer Reviewed
• [Journal Article] Redox Properties of Mononuclear Copper(II)-Superoxide Complex (2013)
  • Author(s): Shinobu Itoh
  • Journal Title: Inorg. Chem. Volume: 52 Issue: 18 Pages: 10431-10437
  • DOI: 10.1021/ic401261z
  • Peer Reviewed
• [Journal Article] Flavonolate Complexes of MII (M = Mn, Fe, Co, Ni, Cu, and Zn). Structural and Functional Models for the ES (Enzyme-Substrate) Complex of Quercetin 2,3-Dioxygenase (2013)
  • Author(s): Shinobu Itoh, et al.
  • Journal Title: Inorg. Chem. Volume: 52 Issue: 19 Pages: 10936-10948
  • DOI: 10.1021/ic400972k
  • Peer Reviewed
• [Presentation] Developing Mononuclear Copper Reaction Site for Oxygen Activation (2015)
  • Author(s): Shinobu Itoh
  • Organizer: Gordon Research Conference (Metals in Biology)
  • Place of Presentation: Ventura, California, USA
  • Year and Date: 2015-01-25 – 2015-01-30
  • Invited
• [Presentation] Oxygen Activation at Mononuclear Copper Reaction Center (2014)
  • Author(s): Shinobu Itoh
  • Organizer: 7th Asian Biological Inorganic Chemistry Conference (AsBIC7)
  • Place of Presentation: Gold Coast, Australia
  • Year and Date: 2014-11-30 – 2014-12-05
  • Invited
• [Presentation] Molecular Oxygen Activation by Copper Proteins and Models (2014)
  • Author(s): Shinobu Itoh
  • Organizer: 9th International Copper Meeting
  • Place of Presentation: Vivo Equense, Italy
  • Year and Date: 2014-10-05 – 2014-10-10
  • Invited
• [Presentation] Controlling Reactivity of Copper-Peroxide Complexes (2014)
  • Author(s): Shinobu Itoh
  • Organizer: 41th International Conference on Coordination Chemistry (ICCC-41)
  • Place of Presentation: Suntec Singapore Convention & Exhibition Center, Singapore
  • Year and Date: 2014-07-21 – 2014-07-25
  • Invited
• [Presentation] Nickel-complex Catalyzed Hydroxylation of Aliphatic and Aromatic Compounds (2014)
  • Author(s): Shinobu Itoh
  • Organizer: 2014 Canada-Japan Workshop (pre-ISHC)
  • Place of Presentation: University of Ottawa, Canada
  • Year and Date: 2014-07-05 – 2014-07-06
  • Invited
• [Presentation] C-H Bond Activation by Copper Active-oxygen Complexes (2014)
  • Author(s): Shinobu Itoh
  • Organizer: International Conference on Hydrogen Atom Transfer (iCHAT 2014)
  • Place of Presentation: Rome, Italy
  • Year and Date: 2014-06-22 – 2014-06-26
  • Invited
• [Presentation] Mononuclear Copper Active Oxygen Complexes Relevant to the Reactive Intermediate Involved in Copper Monooxygenases (2013)
  • Author(s): Shinobu Itoh
  • Organizer: 16th International Conference of Biological Inorganic Chemiastry (ICBIC16)
  • Place of Presentation: Grenoble, France
  • Invited
• [Presentation] Copper Complexes of Non-innocent β-Diketiminate Ligands (2013)
  • Author(s): Shinobu Itoh
  • Organizer: 15th Asian Chemical Congress (15ACC)
  • Place of Presentation: Singapore
  • Invited
• [Presentation] Ligand Design for Oxygen Activation by Copper Complexes (2013)
  • Author(s): Shinobu Itoh
  • Organizer: The 2nd Canada-Japan Joint Symposium on Coordination Chemistry
  • Place of Presentation: Okinawa
  • Invited
• [Presentation] Oxygen Activation Chemistry at Mononuclear Copper Reaction Center (2013)
  • Author(s): Shinobu Itoh
  • Organizer: The 4th Asian Conference on Coordination Chemistry (ACCC4)
  • Place of Presentation: Jeju, South Korea
  • Invited
• [Presentation] Recent Advances in Copper-Dioxygen Chemistry (2013)
  • Author(s): Shinobu Itoh
  • Organizer: Symposium on Modern Trends in Inorganic Chemistry–XV
  • Place of Presentation: Roorkee, India
  • Invited
• [Book] Chemical Science of π-Electron Systems (2015)
  • Author(s): Shinobu Itoh and Yuma Morimoto
  • Total Pages: 500
  • Publisher: Springer
• [Book] Handbook of Metalloproteins to the Encyclopedia of Inorganic and Bioinorganic Chemistry (2015)
  • Author(s): Nobutaka Fujieda and Shinobu Itoh
  • Total Pages: 300
  • Publisher: John Wiley & Sons