津川　若子TSUGAWA Wakakoツガワ　ワカコ

業績情報

氏名・連絡先

• 氏名

  ツガワ　ワカコ, 津川　若子, TSUGAWA Wakako

• 生年

  1963

• 電話番号

  +81-42-388-7789

• Fax番号

  042-388-7027

• eメールアドレス

  tsugawa at cc.tuat.ac.jp

• 個人ホームページ

  http://www.tuat.ac.jp/~tanpaku/

主たる所属・職名

• 工学研究院　生命機能科学部門, 准教授

その他の所属

• 工学府　産業技術専攻
• 工学府　生命工学専攻
• 工学部　生命工学科

経歴

• -
  積水化学工業株式会社綜合研究所メディカル研究所研究員
  自 1987年04月, 至 1992年02月

学歴

• 東京工業大学
  理学部
  化学科
  至 1987年, 卒業

教育・研究活動状況

• 私たちの研究室は酵素などの蛋白質を研究ターゲットの中心において、1)新規な蛋白質を自然界に存在する微生物から単離、2) 構造・機能情報をもとに遺伝子レベルから蛋白質を設計・生産する蛋白質工学的手法による新たな蛋白質の開発、3) さらにこれらの蛋白質の応用として医薬品原料などの新しい物質生産系の開発や医療診断や環境分析などに用いるバイオセンシングシステムの開発、などの研究を進め、新たな産業創出の可能性を探究しています。以下に主な研究成果として、蛋白質工学による糖尿病診断用酵素の開発とバイオセンサーへの応用に関する研究について紹介します。その他の研究活動については、研究室のホームページ(http：//www.tuat.ac.jp/~tanpaku/)をご覧下さい。 蛋白質工学による糖尿病診断バイオセンサー用酵素の開発酵素などの蛋白質は20種類のアミノ酸から構成されていて、その長さや配列により構造を形成し、特徴的な機能を発揮します。蛋白質の構造はすべて遺伝子にコードされていることから、遺伝子の配列を改変して、その改変した遺伝子配列を組み換えDNA技術によって大腸菌などで生産することにより、天然には存在しない優れた機能を有する人工の蛋白質を設計生産できます。このような遺伝子レベルから蛋白質を改良/設計／生産する技術が蛋白質工学とよばれています。私たちが注目している酵素はグルコース脱水素酵素とよばれる蛋白質で、ブドウ糖の酸化を触媒する酵素です。しかし、この酵素は、ブドウ糖以外の似たような構造をもった糖類も酸化します。今後この酵素をブドウ糖分析に応用するためにはブドウ糖だけを選択的に酸化することが必要です。この酵素は約800個のアミノ酸分子から構成されていますが、そのうちの一つのアミノ酸を改変することでこの酵素をブドウ糖とだけ反応する酵素に改良することに成功しました。 酵素を分子認識素子に用いることにより、様々な化合物を極めて高感度に計測することができます。この酵素の機能とエレクトロニクス技術とを融合し、酵素などの生体関連の分子認識素子を用いた計測機器がバイオセンサーと呼ばれています。蛋白質工学によって創られた新しいグルコース脱水素酵素を用いることにより、極めて選択性の高い高感度なブドウ糖測定が行えるバイオセンサーが開発できます。ブドウ糖の測定は血液中のブドウ糖濃度が高くなる糖尿病の診断にはかかせない分析です。このようなバイオセンサーは今後ますます需要が高まってくる家庭での医療診断に基づく健康管理、在宅診断において応用されていくことが期待されています。

研究分野

• A489　ナノテク・材料, A37010　生体化学
• A489　ナノテク・材料, A37010　生体化学

研究キーワード

• 酵素測定法 バイオセンサー 酵素センサー バイオ燃料電池 ポイントオブケア

研究テーマ

• 糖化蛋白質の酵素測定法の開発
  自 2004
• 直接電子移動型グルコース脱水素酵素を用いたグルコース連続測定
  自 9999

担当授業科目

• 産業技術概論
  2022年, 専門科目等
• 産業技術実践研究
  2022年, 専門科目等
• プレゼンテーション実習
  2022年, 専門科目等
• ケーススタディ
  2022年, 専門科目等
• インターンシップ(通年・集中・工学府専門職学位)
  2022年, 専門科目等
• 基礎生物化学(1学期・月4・工学部)
  2022年, 専門科目等
• 食品・医薬品開発工学(3学期・水2・工学部)
  2022年, 専門科目等
• 生命分子産業技術論(1学期・火6・工学府専門職学位)
  2022年, 専門科目等
• 蛋白質・核酸科学(3学期・金1・工学部)
  2022年, 専門科目等

科学研究費助成事業

• 基盤研究(B)
  新奇酵素、鉄硫黄フラボプロテイン・シトクロム脱水素酵素の分子機構の解明
  自 2018年, 至 2018年
• 基盤研究(C)
  グルコシド-3-脱水素酵素オペロン中の機能未知のタンパク質の解析
  自 2017年, 至 2019年
• 基盤研究(B)
  新奇酵素、鉄硫黄フラボプロテイン・シトクロム脱水素酵素の分子機構の解明
  自 2017年, 至 2017年
• 基盤研究(B)
  新奇酵素、鉄硫黄フラボプロテイン・シトクロム脱水素酵素の分子機構の解明
  自 2016年, 至 2016年
• 基盤研究(B)
  ピロロキノリンキノンによるアミロイド蛋白質の選択的構造形成制御
  自 2010年, 至 2010年
• 基盤研究(B)
  ピロロキノリンキノンによるアミロイド蛋白質の選択的構造形成制御
  自 2009年, 至 2009年
• 基盤研究(B)
  ピロロキノリンキノンによるアミロイド蛋白質の選択的構造形成制御
  自 2008年, 至 2008年
• 特定領域研究(公募)
  細胞内蛋白質糖化プロセスのモニタリング
  自 2006年, 至 2006年
• 基盤研究(B)
  酵素・抗体・アプタマーの近接を利用した単一工程電気化学センサーの開発
  自 2023年, 至 2023年

論文

• Structure of lactate oxidase from Enterococcus hirae revealed new aspects of active site loop function: Product-inhibition mechanism and oxygen gatekeeper
  Hiraka, Kentaro; Yoshida, Hiromi; Tsugawa, Wakako; Asano, Ryutaro; La Belle, Jeffrey T.; Ikebukuro, Kazunori; Sode, Koji
  PROTEIN SCIENCE
  WILEY
  l-Lactate oxidase (LOx) is a flavin mononucleotide (FMN)-dependent triose phosphate isomerase (TIM) barrel fold enzyme that catalyzes the oxidation of l-lactate using oxygen as a primary electron acceptor. Although reductive half-reaction mechanism of LOx has been studied by structure-based kinetic studies, oxidative half-reaction and substrate/product-inhibition mechanisms were yet to be elucidated. In this study, the structure and enzymatic properties of wild-type and mutant LOxs from Enterococcus hirae (EhLOx) were investigated. EhLOx structure showed the common TIM-barrel fold with flexible loop region. Noteworthy observations were that the EhLOx crystal structures prepared by co-crystallization with product, pyruvate, revealed the complex structures with d-lactate form ligand, which was covalently bonded with a Tyr211 side chain. This observation provided direct evidence to suggest the product-inhibition mode of EhLOx. Moreover, this structure also revealed a flip motion of Met207 side chain, which is located on the flexible loop region as well as Tyr211. Through a saturation mutagenesis study of Met207, one of the mutants Met207Leu showed the drastically decreased oxidase activity but maintained dye-mediated dehydrogenase activity. The structure analysis of EhLOx Met207Leu revealed the absence of flipping in the vicinity of FMN, unlike the wild-type Met207 side chain. Together with the simulation of the oxygen-accessible channel prediction, Met207 may play as an oxygen gatekeeper residue, which contributes oxygen uptake from external enzyme to FMN. Three clades of LOxs are proposed based on the difference of the Met207 position and they have different oxygen migration pathway from external enzyme to active center FMN.
  2022年10月, 研究論文（学術雑誌）, 共同, 31, 10, 0961-8368, DOI(公開)(r-map)
• Transient potentiometry based D-serine sensor using engineered D-amino acid oxidase showing quasi-direct electron transfer property
  Takamatsu, Shouhei; Lee, Inyoung; Lee, Jinhee; Asano, Ryutaro; Tsugawa, Wakako; Ikebukuro, Kazunori; Dick, Jeffrey E.; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  D-Serine biosensing has been extensively reported based on enzyme sensors using flavin adenine dinucleotide (FAD) -dependent n-amino acid oxidase (DAAOx), based on the monitoring of hydrogen peroxide generated by the enzymatic reaction, which is affected by dissolved oxygen concentration in the measurement environment in in vivo use. Here we report a novel sensing principle for o-serine, transient potentiometry based o-serine sensor using engineered DAAOx showing quasi-direct electron transfer (DET) property. DAAOx Gly52Val mutant, revealed to possess dye-mediated dehydmgenase activity using artificial synthetic electron acceptors, while its oxidase activity was negligible. The enzyme was immobilized on electrode and was modified with amine-reactive phenazine ethosulfate, resulted an enzyme electrode showing quasi-DET type response. Although OCP based monitoring took more than several minutes to obtain steady state OCP value, the time dependent OCP change monitoring, transient potentiometry, provided rapid and sensitive sensor signals. While dOCP/dt based monitoring was suitable for sensing with longer than 5 s time resolution with o-serine concentration range between 0.5 mM and 5 mM, dOCP/d root t based monitoring is suitable for o-serine monitoring with much shorter time resolution (less than 1 s) with high sensitivity with wider dynamic range (20 mu M-30 mM). The maximum dOCP/d root t was -39.2 +/- 2.0 mV/s(1/2), the K-m(app) was 1.9 mM, and the lower limit of detection was 20 mu M. In addition, D-serine monitoring was also possible in the artificial cerebrospinal fluid. The transient potentiometry based sensing reported in this study will be further utilized to realize miniaturized, continuous, real-time, in vivo sensor for o-serine monitoring.
  2022年03月15日, 研究論文（学術雑誌）, 共同, 200, 0956-5663, DOI(公開)(r-map)
• Microgravity environment grown crystal structure information based engineering of direct electron transfer type glucose dehydrogenase
  Okuda-Shimazaki, Junko; Yoshida, Hiromi; Lee, Inyoung; Kojima, Katsuhiro; Suzuki, Nanoha; Tsugawa, Wakako; Yamada, Mitsugu; Inaka, Koji; Tanaka, Hiroaki; Sode, Koji
  COMMUNICATIONS BIOLOGY
  NATURE PORTFOLIO
  The heterotrimeric flavin adenine dinucleotide dependent glucose dehydrogenase is a promising enzyme for direct electron transfer (DET) principle-based glucose sensors within continuous glucose monitoring systems. We elucidate the structure of the subunit interface of this enzyme by preparing heterotrimer complex protein crystals grown under a space microgravity environment. Based on the proposed structure, we introduce inter-subunit disulfide bonds between the small and electron transfer subunits (5 pairs), as well as the catalytic and the electron transfer subunits (9 pairs). Without compromising the enzyme's catalytic efficiency, a mutant enzyme harboring Pro205Cys in the catalytic subunit, Asp383Cys and Tyr349Cys in the electron transfer subunit, and Lys155Cys in the small subunit, is determined to be the most stable of the variants. The developed engineered enzyme demonstrate a higher catalytic activity and DET ability than the wild type. This mutant retains its full activity below 70 ? as well as after incubation at 75 ? for 15 min - much higher temperatures than the current gold standard enzyme, glucose oxidase, is capable of withstanding.
  2022年12月06日, 研究論文（学術雑誌）, 共同, 5, 1, DOI(公開)(r-map)
• A Thiol-reactive Phenazine Ethosulfate - A Novel Redox Mediator for Quasi-direct Electron-transfer-type Sensors
  Fitriana, Maya; Hiraka, Kentaro; Ikebukuro, Kazunori; Sode, Koji; Tsugawa, Wakako
  SENSORS AND MATERIALS
  MYU, SCIENTIFIC PUBLISHING DIVISION
  A novel redox mediator, thiol-reactive phenazine ethosulfate (trPES), was used to modify an enzyme for the first time for biosensor development. Aerococcus viridans-lactate oxidase (LOx), widely used to study lactate biosensors, was modified with a single trPES molecule. A cysteine mutation was introduced into the vicinity of the LOx cofactor to enable the modification by trPES. LOx cysteine mutants were then successfully modified using trPES, thus acquiring quasi-direct electron transfer ability. An electrode immobilized with the trPES-LOx A96L/S210C mutant showed the highest amperometric response currents among the modified LOx cysteine mutants, indicating efficient electron transfer. The position around residues S210 to N212 on the LOx surface (distance <14 angstrom from FMN N5) is important for the mediator to access the reduced flavin. Then, the performances of the lactate sensor were improved by utilizing LOx A96L/S210C modified with trPES and arPES, another redox mediator used to modify a lysine residue. The lactate sensor has a detection range of up to 1 mM, a sensitivity of 6.62 mu A/mM.cm(2), and a limit of detection of 9.9 mu M. Furthermore, Aspergillus flavus-derived FAD glucose dehydrogenase was successfully modified with trPES and the response currents were obtained, showing the versatility of trPES for modifying other oxidoreductases.
  2022年, 研究論文（学術雑誌）, 共同, 34, 6, 0914-4935, DOI(公開)(r-map), 2105, 2124
• Strategic design and improvement of the internal electron transfer of heme b domain-fused glucose dehydrogenase for use in direct electron transfer-type glucose sensors
  Ito, Kohei; Okuda-Shimazaki, Junko; Kojima, Katsuhiro; Mori, Kazushige; Tsugawa, Wakako; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  A fusion enzyme composed of an Aspergillus flavus-derived flavin adenine dinucleotide glucose dehydrogenase (AfGDH) and an electron transfer domain of Phanerochaete chrysosporium-derived cellobiose dehydrogenase (Pcyb) was previously reported to show the direct electron transfer (DET) ability to an electrode. However, its slow intramolecular electron transfer (IET) rate from the FAD to the heme, limited the sensor signals. In this study, fusion FADGDH (Pcyb-AfGDH) enzymes were strategically redesigned by performing docking simulation, following surface-electrostatic potential estimation in the predicted area. Based on these predictions, we selected the amino acid substitution on Glu324, or on Asn408 to Lys to increase the positive charge at the rim of the interdomain region. Pcyb-AfGDH mutants were recombinantly produced using Pichia pastoris as the host microorganism, and their IET was evaluated. Spectroscopic observations showed that the Glu324Lys (E324K) and Asn408Lys (N408K) Pcyb-AfGDH mutants showed approximately 1.70and 9.0-fold faster IET than that of wildtype Pcyb-AfGDH, respectively. Electrochemical evaluation revealed that the mutant Pcyb-AfGDHimmobilized electrodes showed higher DET current values than that of the wildtype Pcyb-AfGDH-immobilized electrodes at pH 6.5, which was approximately 9-fold higher in the E324K mutant and 15-fold higher in the N408K mutant, than in the wildtype. Glucose enzyme sensors employing N408K mutant was able to measure glucose concentration under physiological condition using artificial interstitial fluid at pH 7.4, whereas the one with wildtype Pcyb-AfGDH was not. These results indicated that the sensor employed the redesigned mutant Pcyb-AfGDH can be used for future continuous glucose monitoring system based on direct electron transfer principle. (247 words).
  2021年03月15日, 研究論文（学術雑誌）, 共同, 176, 0956-5663, DOI(公開)(r-map)
• Rational design of direct electron transfer type L-lactate dehydrogenase for the development of multiplexed biosensor
  Hiraka, Kentaro; Tsugawa, Wakako; Asano, Ryutaro; Yokus, Murat A.; Ikebukuro, Kazunori; Daniele, Michael A.; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  The development of wearable multiplexed biosensors has been focused on systems to measure sweat L-lactate and other metabolites, where the employment of the direct electron transfer (DET) principle is expected. In this paper, a fusion enzyme between an engineered L-lactate oxidase derived from Aerococcus viridans, AvLOx A96L/N212K mutant, which is minimized its oxidase activity and b-type cytochrome protein was constructed to realize multiplexed DET-type lactate and glucose sensors. The sensor with a fusion enzyme showed DET to a gold electrode, with a limited operational range less than 0.5 mM. A mutation was introduced into the fusion enzyme to increase K-m value and eliminate its substrate inhibition to construct b2LOxS. Together with the employment of an outer membrane, the detection range of the sensor with b2LOxS was expanded up to 10 mM. A simultaneous lactate and glucose monitoring system was constructed using a flexible thin-film multiplexed electrodes with b2LOxS and a DET-type glucose dehydrogenase, and evaluated their performance in the artificial sweat. The sensors achieved simultaneous detection of lactate and glucose without cross-talking error, with the detected linear ranges of 0.5-20 mM for lactate and 0.1-5 mM for glucose, sensitivities of 4.1 nA/mM.mm(2) for lactate and 56 nA/mM.mm(2) for glucose, and limit of detections of 0.41 mM for lactate and 0.057 mM for glucose. The impact of the presence of electrochemical interferants (ascorbic acid, acetaminophen and uric acid), was revealed to be negligible. This is the first report of the DET-type enzyme based lactate and glucose dual sensing systems.
  2021年03月15日, 研究論文（学術雑誌）, 共同, 176, 0956-5663, DOI(公開)(r-map)
• Development of glycated peptide enzyme sensor based flow injection analysis system for haemoglobin A1c monitoring using quasi-direct electron transfer type engineered fructosyl peptide oxidase
  Hatada, Mika; Saito, Satomi; Yonehara, Satoshi; Tsugawa, Wakako; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  Haemoglobin A1e (hemoglobin A1e, HbA1c) is an important long-term glycemic control marker for diabetes. The aim of this study was to develop an enzyme flow injection analysis (FIA) system using engineered fructosyl peptide oxidase (FPOx) based on 2.5th generation principle for an HbA1c automated analytical system. FPOx from Phaeosphaeria nodorum (PnFPOx) was engineered by introducing a Lys residue at the R414 position, to be modified with amine reactive phenazine ethosulfate (arPES) in proximity of FAD. The engineered PnFPOx mutant with minimized oxidase activity, N56A/R414K, showed quasi-direct electron transfer (quasi-DET) ability after PES-modification. The FIA system was constructed by employing a PES-modified PnFPOx N56A/R414K and operated at 0 V against Ag/AgCl. The system showed reproducible responses with a linear range of 20-500 mu M for both fructosyl valine (FV) and fructosyl valylhistidine (FVH), with sensitivities of 0.49 nA mu AO and 0.13 nA mu M-1, and the detection limits of 1.3 mu M and 2.0 mu M for FV and FVH, respectively. These results indicate that the enzyme electrochemical FIA system covers the clinical range of HbA1c detection for more 200 consecutive measurements. Protease digested three different levels of HbA1c samples including healthy and diabetic range subjects were also measured with the FIA system. Thus, it will be possible to develop an integrated system consisting of sample pretreatment and sample electrochemical measurement based on an FIA system possessing quasi-DET type PnFPOx.
  2021年04月01日, 研究論文（学術雑誌）, 共同, 177, 1, 0956-5663, DOI(公開)(r-map), 112984
• Development of an Interdigitated Electrode-Based Disposable Enzyme Sensor Strip for Glycated Albumin Measurement
  Hatada, Mika; Loew, Noya; Okuda-Shimazaki, Junko; Khanwalker, Mukund; Tsugawa, Wakako; Mulchandani, Ashok; Sode, Koji
  MOLECULES
  MDPI
  Glycated albumin (GA) is an important glycemic control marker for diabetes mellitus. This study aimed to develop a highly sensitive disposable enzyme sensor strip for GA measurement by using an interdigitated electrode (IDE) as an electrode platform. The superior characteristics of IDE were demonstrated using one microelectrode of the IDE pair as the working electrode (WE) and the other as the counter electrode, and by measuring ferrocyanide/ferricyanide redox couple. The oxidation current was immediately reached at the steady state when the oxidation potential was applied to the WE. Then, an IDE enzyme sensor strip for GA measurement was prepared. The measurement of fructosyl lysine, the protease digestion product of GA, exhibited a high, steady current immediately after potential application, revealing the highly reproducible measurement. The sensitivity (2.8 nA mu M-1) and the limit of detection (1.2 mu M) obtained with IDE enzyme sensor strip were superior compared with our previously reported sensor using screen printed electrode. Two GA samples, 15 or 30% GA, corresponding to healthy and diabetic levels, respectively, were measured after protease digestion with high resolution. This study demonstrated that the application of an IDE will realize the development of highly sensitive disposable-type amperometric enzyme sensors with high reproducibility.
  2021年02月, 研究論文（学術雑誌）, 共同, 26, 3, DOI(公開)(r-map), 734
• Third generation impedimetric sensor employing direct electron transfer type glucose dehydrogenase
  Ito, Yuka; Okuda-Shimazaki, Junko; Tsugawa, Wakako; Loew, Noya; Shitanda, Isao; Lin, Chi-En; La Belle, Jeffrey; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  Faradaic electrochemical impedance spectroscopy (faradaic EIS) is an attractive measurement principle for biosensors. However, there have been no reports on sensors employing direct electron transfer (DET)-type redox enzymes based on faradaic EIS principle. In this study, we have attempted to construct the 3rd-generation faradaic enzyme EIS sensor, which used DET-type flavin adenine dinucleotide (FAD) dependent glucose dehydrogenase (GDH) complex, to elucidate its characteristic properties as well as to investigate its potential application as the future immunosensor platform. The gold disk electrodes (GDEs) with DET-type FADGDH prepared using self-assembled monolayer (SAM) showed the glucose concentration dependent impedance change, which was confirmed by the change in the charge transfer resistance (R-ct). The Delta(1/R-ct) values were also affected by DC bias potential and the length of SAM. Based on the Nyquist plot and Bode plot simulations, glucose sensing by imaginary impedance monitoring under fixed frequency (5 mHz) was carried out, revealing the higher sensitivity at low glucose concentration with wider linear range (0.024-0.2 mM). Considering this high sensitivity toward glucose, the 3rd-generation faradaic enzyme EIS sensor would provide alternative platform for future impedimetric immunosensing system, which does not use redox probe.
  2019年03月15日, 研究論文（学術雑誌）, 共同, 129, 0956-5663, DOI(公開)(r-map), 189, 197
• Affinity sensor for haemoglobin A1c based on single-walled carbon nanotube field-effect transistor and fructosyl amino acid binding protein
  Hatada, Mika; Thien-Toan Tran; Tsugawa, Wakako; Sode, Koji; Mulchandani, Ashok
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  Haemoglobin A1c (HbA1c) is a significant glycaemic marker for diabetes mellitus. The level of HbA1c reflects the mean blood glucose level over the prior 2-3 months and it is useful for the assessment of therapeutic effectiveness and for diagnosis. In this study, we report the label-free affinity sensor for HbA1c based on the chemiresistor-type field-effect transistor, which has a simple sensor configuration. Single-walled carbon nano tubes (SWNTs) were used as the transducing element. The fructosyl amino acid binding protein from Rhizobiwn radiobacter (SocA), which binds to a-fructosyl amino acid specifically, was used as the biorecognition element for fructosyl valine (FV), the product of the proteolytic hydrolysis of HbA1c. The developed sensor shows the ability to measure as low as L2 nM FV, which is 14-fold more sensitive compared to the previously reported fluorescence -based sensor using SocA. This sensor also exhibits high specificity where no significant response is observed from either fructosyl lysine (FK) or glucose, which are potential interferents. FK is the alpha-fructosyl amino acid from glycated albumin, another glycated protein, whereas glucose is naturally present at very high concentration in the blood. We propose that the modulation of the surface charges on the SWNTs caused by the conformational change in SocA upon ligand binding leads to the proportionate changes in the number of carriers in the SWNT channel.
  2019年03月15日, 研究論文（学術雑誌）, 共同, 129, 0956-5663, DOI(公開)(r-map), 254, 259
• Development of a third-generation glucose sensor based on the open circuit potential for continuous glucose monitoring
  Lee, Inyoung; Loew, Noya; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  Continuous glucose monitoring (CGM) systems are most important in the current Type I diabetes care and as component for the development of artificial pancreas systems because the amount of insulin being supplied is calculated based on the CGM results. Therefore, to stably and accurately control the blood glucose level, CGM should be stable and accurate for a long period. We have been engaged in the biomolecular engineering and application of FAD dependent glucose dehydrogenase complex (FADGDH) which is capable of direct electron transfer. In this study, we report the development of the third-generation type open circuit potential (OCP) principle-based glucose sensor with direct electron transfer FADGDH immobilized on gold electrodes using a self-assembled monolayer (SAM). We developed a novel algorithm for OCP-based glucose sensors. By employing this new algorithm, high reproducibility of measurement and sensor preparation were achieved. In addition, the signal was not affected by the presence of acetaminophen and ascorbic acid in the sample solution. The thus optimized third-generation OCP-based glucose sensor could be operated continuously for more than 9 days without significant change in the signal, sensitivity and dynamic range, indicating its potential application for CGM systems.
  2019年01月15日, 研究論文（学術雑誌）, 共同, 124, 0956-5663, DOI(公開)(r-map), 216, 223
• Designer fungus FAD glucose dehydrogenase capable of direct electron transfer
  Ito, Kohei; Okuda-Shimazaki, Junko; Mori, Kazushige; Kojima, Katsuhiro; Tsugawa, Wakako; Ikebukuro, Kazunori; Lin, Chi-En; La Belle, Jeffrey; Yoshida, Hiromi; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  Fungi-derived flavin adenine dinucleotide glucose dehydrogenases (FADGDHs) are currently the most popular and advanced enzymes for self-monitoring of blood glucose sensors; however, the achievement of direct electron transfer (DET) with FADGDHs is difficult. In this study, a designer FADGDH was constructed by fusing Aspergillus flavus derived FADGDH (AfGDH) and a Phanerochaete chrisosporium CDH (PcCDH)-derived heme b-binding cytochrome domain to develop a novel FADGDH that is capable of direct electron transfer with an electrode. A structural prediction suggested that the heme in the CDH may exist in proximity to the FAD of AfGDH if the heme b-binding cytochrome domain is fused to the AfGDH N-terminal region. Spectroscopic observations of recombinantly produced designer FADGDH confirmed the intramolecular electron transfer between FAD and the heme. A decrease in pH and the presence of a divalent cation improved the intramolecular electron transfer. An enzyme electrode with the immobilized designer FADGDH showed an increase in current immediately after the addition of glucose in a glucose concentration-dependent manner, whereas those with wild-type AfGDH did not show an increase in current. Therefore, the designer FADGDH was confirmed to be a novel GDH that possesses electrode DET ability. The difference in the surface electrostatic potentials of AfGDH and the catalytic domain of PcCDH might be why their intramolecular electron transfer ability is inferior to that of CDH. These relevant and consistent findings provide us with a novel strategic approach for the improvement of the DET properties of designer FADGDH. (241 words)
  2019年01月01日, 研究論文（学術雑誌）, 共同, 123, 0956-5663, DOI(公開)(r-map), 114, 123
• X-ray structure of the direct electron transfer-type FAD glucose dehydrogenase catalytic subunit complexed with a hitchhiker protein
  Hiromi Yoshida, Katsuhiro Kojima, Masaki Shiota, Keiichi Yoshimatsu, Tomohiko Yamazaki, Stefano Ferri, Wakako Tsugawa, Shigehiro Kamitori, Koji Sode
  ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
  INT UNION CRYSTALLOGRAPHY
  The bacterial flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase complex derived from Burkholderia cepacia (BcGDH) is a representative molecule of direct electron transfer-type FAD-dependent dehydrogenase complexes. In this study, the X-ray structure of BcGDH gamma alpha, the catalytic subunit (alpha-subunit) of BcGDH complexed with a hitchhiker protein (gamma-subunit), was determined. The most prominent feature of this enzyme is the presence of the 3Fe-4S cluster, which is located at the surface of the catalytic subunit and functions in intramolecular and intermolecular electron transfer from FAD to the electron-transfer subunit. The structure of the complex revealed that these two molecules are connected through disulfide bonds and hydrophobic interactions, and that the formation of disulfide bonds is required to stabilize the catalytic subunit. The structure of the complex revealed the putative position of the electron-transfer subunit. A comparison of the structures of BcGDH gamma alpha and membrane-bound fumarate reductases suggested that the whole BcGDH complex, which also includes the membrane-bound beta-subunit containing three heme c moieties, may form a similar overall structure to fumarate reductases, thus accomplishing effective electron transfer.
  2019年08月29日, 研究論文（学術雑誌）, 共同, 75, Pt9, DOI(公開)(r-map), 841, 851
• G-Quadruplex Structure Improves the Immunostimulatory Effects of CpG Oligonucleotides.
  Kazuaki Hoshi, Tomohiko Yamazaki, Yuuki Sugiyama, Kaori Tsukakoshi, Wakako Tsugawa, Koji Sode, Kazunori Ikebukuro
  Nucleic acid therapeutics
  Single-strand oligodeoxynucleotides (ODNs) containing unmethylated cytosine-phosphate-guanine (CpG) are recognized by the toll-like receptor 9, a component of the innate immunity. Therefore, they could act as immunotherapeutic agents. Chemically modified CpG ODNs containing a phosphorothioate backbone instead of phosphodiester (PD) were developed as immunotherapeutic agents resistant to nuclease degradation. However, they cause adverse side effects, and so there is a necessity to generate novel CpG ODNs. In the present study, we designed a nuclease-resistant nonmodified CpG ODN that forms G-quadruplex structures. G-quadruplex formation in CpG ODNs increased nuclease resistance and cellular uptake. The CpG ODNs designed in this study induced interleukin-6 production in a human B lymphocyte cell line and human peripheral blood mononuclear cells. These results indicate that G-quadruplex formation can be used to increase the immunostimulatory activity of CpG ODNs having a natural PD backbone.
  2019年08月06日, 研究論文（学術雑誌）, 共同, 29, 4, DOI(公開)(r-map), 224, 229
• Synthesis of a hemin-containing copolymer as a novel immunostimulator that induces IFN-gamma production
  Hoshi, Kazuaki; Yamazaki, Tomohiko; Yoshikawa, C. Hiaki; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji
  INTERNATIONAL JOURNAL OF NANOMEDICINE
  DOVE MEDICAL PRESS LTD
  Background: Hemozoin, a chemical analog of a malarial pigment, is a crystal composed of heme dimers that can act as a potent Th1-type adjuvant, which strongly induces antibody production. However, the clinical applications of malarial hemozoin have limitations due to biosafety concerns and difficulties in the manufacturing process. Based on the premise that an analog of the heme polymer might display immunostimulatory effects, a hemin-containing polymer was developed as a novel immunostimulator. Materials and methods: To synthesize the copolymer containing hemin and N-isopropylacrylamide (NIPAM), this study employed a conventional radical polymerization method using 2,2'-azodiisobutyronitrile as the radical initiator; the synthesized copolymer was designated as NIPAM-hemin. Results: NIPAM-hemin was soluble and showed no cytotoxicity in vitro. The NIPAM-hemin copolymer induced the production of interferon (IFN)-gamma and interleukin (IL)-6 from peripheral blood mononuclear cells, although hemin and the NIPAM monomer individually did not induce the production of any cytokines. The production of IFN-gamma induced by NIPAM-hemin was independent of toll-like receptor 9 and the NLRP3 inflammasome pathway. Conclusion: Given that NIPAM-hemin induced IL-6 and IFN-gamma production in immune cells without any cytotoxic effects, NIPAM-hemin has potential therapeutic applications as a Th1-type adjuvant.
  2018年, 研究論文（学術雑誌）, 共同, 13, 1178-2013, DOI(公開)(r-map), 4461, 4472
• Convenient and Universal Fabrication Method for Antibody-Enzyme Complexes as Sensing Elements Using the SpyCatcher/SpyTag System
  Kimura, Hayato; Asano, Ryutaro; Tsukamoto, Natsumi; Tsugawa, Wakako; Sode, Koji
  ANALYTICAL CHEMISTRY
  AMER CHEMICAL SOC
  Antibody enzyme complexes (AECs) are ideal sensing elements, especially when oxidoreductases are used as the enzymes in the complex, with the potential to carry out rapid electrochemical measurements. However, conventional methods for the fabrication of AECs, including direct fusion and chemical conjugation, are associated with issues regarding the generation of insoluble aggregates and production of homogeneous AECs. Here, we developed a convenient and universal method for the fabrication of homogeneous AECs using the SpyCatcher/SpyTag system. We used an anti-epidermal growth factor receptor (EGFR) variable domain of a heavy chain antibody (VHH) and a glucose dehydrogenase (GDH) derived from Aspergillus flavus (AfGDH) as the model antibody and enzyme, respectively. Both SpyTag-fused VHH and SpyCatcher-fused AfGDH were successfully prepared using an Escherichia coli expression system, whereas anti-EGFR AECs were produced by simply mixing the two fusion proteins. A bivalent AEC, Af GDH with two VHH at both terminals, was also prepared and exhibited an increased affinity. A soluble EGFR was successfully detected in a dose-dependent manner using immobilized anti-EGFR immunoglobulin G (IgG) and bivalent AEC. We also confirmed the universality of this AEC fabricating method by applying it to another VHH. This method results in the convenient and universal preparation of sensing elements with the potential for electrochemical measurement.
  2018年12月18日, 研究論文（学術雑誌）, 共同, 90, 24, 0003-2700, DOI(公開)(r-map), 14500, 14506
• A Designed L-Lactate Dehydrogenase Derived from L-Lactate Oxidase by Oxygen Accessible Pathway Engineering
  Hiraka, Kentaro; Kojima, Katsuhiro; Lin, Chi-En; Tsugawa, Wakako; Asano, Ryutaro; Yoshida, Hiromi; La Belle, Jeffrey; Sode, Koji
  PROTEIN SCIENCE
  WILEY
  2018年11月, 研究論文（学術雑誌）, 共同, 27, 0961-8368, DOI(公開)(r-map), 137, 138
• Engineered fungus derived FAD-dependent glucose dehydrogenase with acquired ability to utilize hexaammineruthenium(III) as an electron acceptor
  Okurita, Madoka; Suzuki, Nanami; Loew, Noya; Yoshida, Hiromi; Tsugawa, Wakako; Mori, Kazushige; Kojima, Katsuhiro; Klonoff, David C.; Sode, Koji
  BIOELECTROCHEMISTRY
  ELSEVIER SCIENCE SA
  Fungal FAD-dependent glucose dehydrogenases (FADGDHs) are considered to be superior enzymes for glucose sensor strips because of their insensitivity to oxygen and maltose. One highly desirable mediator for enzyme sensor strips is hexaammineruthenium(III) chloride because of its low redox potential and high storage stability. However, in contrast to glucose oxidase (GOx), fungal FADGDH cannot utilize hexaammineruthenium(III) as electron acceptor. Based on strategic structure comparison between FADGDH and GOx, we constructed a mutant of Aspergillus flavus-derived FADGDH, capable of utilizing hexaammineruthenium(III) as electron acceptor: AfGDH-H403D. In AfGDH-H403D, a negative charge introduced at the pathway-entrance leading to the FAD attracts the positively charged hexaammineruthenium(III) and guides it into the pathway. The corresponding amino acid in wild-type GOx is negatively charged, which explains the ability of GOx to utilize hexaammineruthenium(III) as electron acceptor. Electrochemical measurements showed a response current of 46.0 mu A for 10 mM glucose with AfGDH-H403D and hexaammineruthenium(III), similar to that with wild-type AfGDH and ferricyanide (47.8 mu A). Therefore, AfGDH-H403D is suitable for constructing enzyme electrode strips with hexaammineruthenium(III) chloride as sole mediator. Utilization of this new, improved fungal FADGDH should lead to the development of sensor strips for blood glucose monitoring with increased accuracy and less stringent packing requirements. (C) 2018 Elsevier B.V. All rights reserved.
  2018年10月, 研究論文（学術雑誌）, 共同, 123, 1567-5394, DOI(公開)(r-map), 62, 69
• Elucidation of the intra- and inter-molecular electron transfer pathways of glucoside 3-dehydrogenase
  Miyazaki, Ryota; Yamazaki, Tomohiko; Yoshimatsu, Keiichi; Kojima, Katsuhiro; Asano, Ryutaro; Sode, Koji; Tsugawa, Wakako
  BIOELECTROCHEMISTRY
  ELSEVIER SCIENCE SA
  Glucoside 3 dehydrogenase (G3DH) is a flavin adenine dinucleotide (FAD)-containing oxidoreductase that catalyzes the oxidation of the hydroxy group on the C-3 position of pyranose and shows broad substrate specificity by oxidizing many saccharides. Due to unique site specificity and wide substrate specificity, G3DHs can be used for synthesis of sugar derivatives, anodic catalysis in biofuel cells, multi-sugar analysis using enzyme electrode, and for enzymatic detection of 1,5 anhydro D glucitol, a clinical marker for diabetes. However, few studies have focused on the fundamental biochemical properties of G3DH, including its electron transfer pathway. In this study, we isolated the G3DH gene from Rhizobium radiobacter, a homologue of marine bacterial G3DH, and reported that the isolated gene fragment contains the genes encoding the G3DH catalytic subunit (subunit I), G3DH hitch-hiker subunit (subunit II), and cytochrome c-like molecule (CYTc). Furthermore, we report the recombinant expression of G3DH from R. radiobacter in Escherichia coli, the characterization of recombinant G3DH and the investigation of the molecular electron pathway of G3DH. We first prepared the G3DH subunit I-II complex using a co-expression vector for both subunits. The G3DH subunit I-II complex showed dye mediated G3DH activity toward methyl a D glucoside (MaG). Electron paramagnetic resonance (EPR) and inductively coupled plasma optical emission spectroscopy (ICP-OES) analyses revealed that subunit I contains an iron-sulfur cluster. We, then, prepared recombinant CYTc and revealed that it is capable of accepting electrons from the catalytic subunit of G3DH by absorption spectrum analysis. These results suggested that R. radiobacter G3DH possesses an iron sulfur cluster that may play an important role in the electron transfer from FAD to cytochrome c like molecule, which is an external electron acceptor of G3DH. Furthermore, we demonstrated that CYTc mediate the electron transfer from G3DH to electrode without the artificial electron mediator. (C) 2018 Elsevier B.V. All rights reserved.
  2018年08月, 研究論文（国際会議プロシーディングス）, 共同, 122, 1567-5394, DOI(公開)(r-map), 115, 122
• Construction and characterization of flavin adenine dinucleotide glucose dehydrogenase complex harboring a truncated electron transfer subunit
  Okuda-Shimazaki, Junko; Loew, Noya; Hirose, Nana; Kojima, Katsuhiro; Mori, Kazushige; Tsugawa, Wakako; Sode, Koji
  ELECTROCHIMICA ACTA
  PERGAMON-ELSEVIER SCIENCE LTD
  One of the most prominent glucose dehydrogenases (GDHs) capable of direct electron transfer with electrodes is the FADGDH complex derived from Burkholderia cepacia. This FADGDH complex comprises the following three distinct subunits: the catalytic subunit (a subunit) that has an FAD cofactor in its redox center, a hitch-hiker protein from the bacterial TAT secretion system (g subunit), and the electron transfer subunit (b subunit). The electron transfer subunit (b subunit) of the FADGDH complex is a threeheme c containing cytochrome c like molecule (heme 1, heme 2 and heme 3 from the N-terminal). In this study, an FADGDH complex harboring a truncated electron transfer subunit composed of only heme 3 was constructed, and its enzymatic activity and electrochemical properties were investigated to elucidate the role of heme 3 and its region. A truncated electron transfer subunit, trb subunit, was designed using the 3D structures of homologous cytochrome c proteins. The designed trb subunit was expressed as soluble and functional heme c molecules forming complexes with ga catalytic complexes. Thus, the formed FADGDH complex has inter-molecular electron transfers from the FAD to the trb subunit, and from the trb subunit to the external electron acceptor, showing electron transfer subunit-mediated characteristic dye-mediated dehydrogenase activity with a Ru-complex. Therefore, heme 3 in the electron transfer subunit is responsible for accepting the electron from the ga catalytic complex. Moreover, the FADGDH complex harboring the trb subunit showed DET activity toward the electrode. Spectroelectrochemical observations revealed that the trb subunit possessed a lower formal potential than any of the 3 hemes in the whole electron transfer subunit. These unexpected electrochemical properties of the heme in the trb subunit may potentially result in the construction of a DET principle-based glucose sensor, which can be operated at a much lower potential than those achieved using the FADGDH complex with a whole electron transfer subunit. (C) 2018 Published by Elsevier Ltd.
  2018年07月01日, 研究論文（学術雑誌）, 共同, 277, 0013-4686, DOI(公開)(r-map), 276, 286
• Development of a glucose sensor employing quick and easy modification method with mediator for altering electron acceptor preference
  Hatada, Mika; Loew, Noya; Inose-Takahashi, Yuka; Okuda-Shimazaki, Junko; Tsugawa, Wakako; Mulchandani, Ashok; Sode, Koji
  BIOELECTROCHEMISTRY
  ELSEVIER SCIENCE SA
  Enzyme based electrochemical biosensors are divided into three generations according to their type of electron transfer from the cofactors of the enzymes to the electrodes. Although the 3rd generation sensors using direct electron transfer (DET) type enzymes are ideal, the number of enzyme types which possess DET ability is limited. In this study, we report of a glucose sensor using mediator-modified glucose dehydrogenase (GDH), that was fabricated by a new quick-and-easy method using the pre-functionalized amine reactive phenazine ethosulfate (arPES). Thus mediator-modified GDH obtained the ability to transfer electrons to bulky electron acceptors as well as electrodes. The concentration of glucose was successfully measured using electrodes with immobilized PES-modified GDH, without addition of external electron mediators. Therefore, continuous monitoring systems can be developed based on this ”2.5th generation” electron transfer principle utilizing quasi-DET. Furthermore, we successfully modified two other diagnostically relevant enzymes, glucoside 3-dehydrogenase and lactate oxidase, with PES. Therefore, various kinds of diagnostic enzymes can achieve quasi-DET ability simply by modification with arPES, suggesting that continuous monitoring systems based on the 2.5th generation principle can be developed for various target molecules. (C) 2018 Elsevier B.V. All rights reserved.
  2018年06月, 研究論文（学術雑誌）, 共同, 121, 1567-5394, DOI(公開)(r-map), 185, 190
• The electrochemical behavior of a FAD dependent glucose dehydrogenase with direct electron transfer subunit by immobilization on self-assembled monolayers
  Lee, Inyoung; Loew, Noya; Tsugawa, Wakako; Lin, Chi-En; Probst, David; La Belle, Jeffrey T.; Sode, Koji
  BIOELECTROCHEMISTRY
  ELSEVIER SCIENCE SA
  Continuous glucose monitoring (CGM) is a vital technology for diabetes patients by providing tight glycemic control. Currently, many commercially available CGM sensors use glucose oxidase (GOD) as sensor element, but this enzyme is not able to transfer electrons directly to the electrode without oxygen or an electronic mediator. We previously reported a mutated FAD dependent glucose dehydrogenase complex (FADGDH) capable of direct electron transfer (DET) via an electron transfer subunit without involving oxygen or a mediator. In this study, we investigated the electrochemical response of DET by controlling the immobilization of DET-FADGDH using 3 types of self-assembled monolayers (SAMs) with varying lengths. With the employment of DET-FADGDH and SAM, high current densities were achieved without being affected by interfering substances such as acetaminophen and ascorbic acid. Additionally, the current generated from DET-FADGDH electrodes decreased with increasing length of SAM, suggesting that the DET ability can be affected by the distance between the enzyme and the electrode. These results indicate the feasibility of controlling the immobilization state of the enzymes on the electrode surface. (C) 2017 Published by Elsevier B.V.
  2018年06月, 研究論文（学術雑誌）, 共同, 121, 1567-5394, DOI(公開)(r-map), 1, 6
• Mutagenesis Study of the Cytochrome c Subunit Responsible for the Direct Electron Transfer-Type Catalytic Activity of FAD-Dependent Glucose Dehydrogenase
  Yamashita, Yuki; Suzuki, Nanoha; Hirose, Nana; Kojima, Katsuhiro; Tsugawa, Wakako; Sode, Koji
  INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
  MDPI
  The FAD-dependent glucose dehydrogenase from Burkholderia cepacia (FADGDH) is a hetero-oligomeric enzyme that is capable of direct electron transfer (DET) with an electrode. The cytochrome c (cyt c) subunit, which possesses three hemes (heme 1, heme 2, and heme 3, from the N-terminal sequence), is known to enable DET; however, details of the electron transfer pathway remain unknown. A mutagenesis investigation of the heme axial ligands was carried out to elucidate the electron transfer pathway to the electron mediators and/or the electrode. The sixth axial ligand for each of the three heme irons, Met109, Met263, and Met386 were substituted with His. The catalytic activities of the wild-type (WT) and mutant enzymes were compared by investigating their dye-mediated dehydrogenase activities and their DET abilities toward the electrode. The results suggested that (1) heme 1 with Met109 as an axial ligand is mainly responsible for the electron transfer with electron acceptors in the solution, but not for the DET with the electrode; (2) heme 2 with Met263 is responsible for the DET-type reaction with the electrode; and (3) heme 3 with Met386 seemed to be the electron acceptor from the catalytic subunit. From these results, two electron transfer pathways were proposed depending on the electron acceptors. Electrons are transferred from the catalytic subunit to heme 3, then to heme 2, to heme 1 and, finally, to electron acceptors in solution. However, if the enzyme complex is immobilized on the electrode and is used as electron acceptors, electrons are passed to the electrode from heme 2.
  2018年04月, 研究論文（学術雑誌）, 共同, 19, 4, 1422-0067, DOI(公開)(r-map)
• Minimizing the effects of oxygen interference on L-lactate sensors by a single amino acid mutation in Aerococcus viridans L-lactate oxidase
  Hiraka, Kentaro; Kojima, Katsuhiro; Lin, Chi-En; Tsugawa, Wakako; Asano, Ryutaro; La Belle, Jeffrey T.; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  L-lactate biosensors employing L-lactate oxidase (LOx) have been developed mainly to measure L-lactate concentration for clinical diagnostics, sports medicine, and the food industry. Some L-lactate biosensors employ artificial electron mediators, but these can negatively impact the detection of L-lactate by competing with the primary electron acceptor: molecular oxygen. In this paper, a strategic approach to engineering an AvLOx that minimizes the effects of oxygen interference on sensor strips was reported. First, we predicted an oxygen access pathway in Aerococcus viridans LOx (AvLOx) based on its crystal structure. This was subsequently blocked by a bulky amino acid substitution. The resulting Ala96Leu mutant showed a drastic reduction in oxidase activity using molecular oxygen as the electron acceptor and a small increase in dehydrogenase activity employing an artificial electron acceptor. Secondly, the Ala96Leu mutant was immobilized on a screen-printed carbon electrode using glutaraldehyde cross-linking method. Amperometric analysis was performed with potassium ferricyanide as an electron mediator under argon or atmospheric conditions. Under argon condition, the response current increased linearly from 0.05 to 0.5 mM L-lactate for both wild-type and Ala96Leu. However, under atmospheric conditions, the response of wild-type AvLOx electrode was suppressed by 9-12% due to oxygen interference. The Ala96Leu mutant maintained 56-69% of the response current at the same L-lactate level and minimized the relative bias error to 19% from 49% of wild-type. This study provided significant insight into the enzymatic reaction mechanism of AvLOx and presented a novel approach to minimize oxygen interference in sensor applications, which will enable accurate detection of L-lactate concentrations.
  2018年04月30日, 研究論文（学術雑誌）, 共同, 103, 0956-5663, DOI(公開)(r-map), 163, 170
• Characterization of Electron Mediator Preference of Aerococcus viridans-Derived Lactate Oxidase for Use in Disposable Enzyme Sensor Strips
  Noya Loew, Maya Fitriana, Kentaro Hiraka, Koji Sode] and Wakako Tsugawa
  Sensors and Materials
  2017年12月, 研究論文（学術雑誌）, 共同, 29, 12, 1703, 1711
• Minimally Invasive Microneedle Array Electrodes Employing Direct Electron Transfer Type Glucose Dehydrogenase for the Development of Continuous Glucose Monitoring Sensors
  SanjivSharmaaEriTakagibTonyCassaWakakoTsugawabKojiSode
  Procedia Technology
  2017年10月, 研究論文（学術雑誌）, 共同, 27, -, 208, 209
• Continuous operation of an ultra-low-power microcontroller using glucose as the sole energy source
  Lee, Inyoung; Sode, Takashi; Loew, Noya; Tsugawa, Wakako; Lowe, Christopher Robin; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  An ultimate goal for those engaged in research to develop implantable medical devices is to develop mechatronic implantable artificial organs such as artificial pancreas. Such devices would comprise at least a sensor module, an actuator module, and a controller module. For the development of optimal mechatronic implantable artificial organs, these modules should be self-powered and autonomously operated. In this study, we aimed to develop a microcontroller using the BioCapacitor principle. A direct electron transfer type glucose dehydrogenase was immobilized onto mesoporous carbon, and then deposited on the surface of a miniaturized Au electrode (7 mm(2)) to prepare a miniaturized enzyme anode. The enzyme fuel cell was connected with a 100 mu F capacitor and a power boost converter as a charge pump. The voltage of the enzyme fuel cell was increased in a stepwise manner by the charge pump from 330 mV to 3.1 V, and the generated electricity was charged into a 100 mu F capacitor. The charge pump circuit was connected to an ultra-low-power microcontroller. Thus prepared BioCapacitor based circuit was able to operate an ultra-low-power microcontroller continuously, by running a program for 17 h that turned on an LED every 60 s. Our success in operating a microcontroller using glucose as the sole energy source indicated the probability of realizing implantable self-powered autonomously operated artificial organs, such as artificial pancreas.
  2017年07月, 研究論文（学術雑誌）, 共同, 93, 0956-5663, DOI(公開)(r-map), 335, 339
• Development of a screen-printed carbon electrode based disposable enzyme sensor strip for the measurement of glycated albumin
  Hatada, Mika; Tsugawa, Wakako; Kamio, Eri; Loew, Noya; Klonoff, David C.; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  Glycated proteins, such as glycated hemoglobin (HbA1c) or glycated albumin (GA) in the blood, are essential indicators of glycemic control for diabetes mellitus. Since GA, compared to HbA1c, is more sensitive to short term changes in glycemic levels, GA is expected to be used as an alternative or together with HbA1c as a surrogate marker indicator for glycemic control. In this paper we report the development of a sensing system for measuring GA by combining an enzyme analysis method, which is already used in clinical practice, with electrochemical principles. We used fructosyl amino acid oxidase, hexaammineruthenium(III) chloride as the electron mediator, and an inexpensive and economically attractive screen-printed carbon electrode. We used chronoamperometry to measure protease-digested GA samples. The developed sensor strips were able to measure protease-digested samples containing GA in very small sample volumes (1.3 mu L) within about 1 min. We also prepared enzyme sensor strips suitable for clinical use in which the enzyme and the mediator were deposited and dried on. This sensor system showed a clear correlation between the GA concentration and the resulting current. The strips were stable following 3 months of storage at 37 degrees C. We conclude that this disposable enzyme sensor strip system for measuring GA is suitable for point-of-care test (POCT) applications. (C) 2016 Elsevier B.V. All rights reserved.
  2017年02月, 研究論文（国際会議プロシーディングス）, 共同, 88, 0956-5663, DOI(公開)(r-map), 167, 173
• Mediator Preference of Two Different FAD-Dependent Glucose Dehydrogenases Employed in Disposable Enzyme Glucose Sensors
  Noya Loew, Wakako Tsugawa, Daichi Nagae, Katsuhiro Kojima, Koji Sode
  SENSORS
  MDPI AG
  Most commercially available electrochemical enzyme sensor strips for the measurement of blood glucose use an artificial electron mediator to transfer electrons from the active side of the enzyme to the electrode. One mediator recently gaining attention for commercial sensor strips is hexaammineruthenium(III) chloride. In this study, we investigate and compare the preference of enzyme electrodes with two different FAD-dependent glucose dehydrogenases (FADGDHs) for the mediators hexaammineruthenium(III) chloride, potassium ferricyanide (the most common mediator in commercial sensor strips), and methoxy phenazine methosulfate (mPMS). One FADGDH is a monomeric fungal enzyme, and the other a hetero-trimeric bacterial enzyme. With the latter, which contains a heme-subunit facilitating the electron transfer, similar response currents are obtained with hexaammineruthenium(III), ferricyanide, and mPMS (6.8 mu A, 7.5 mu A, and 6.4 mu A, respectively, for 10 mM glucose). With the fungal FADGDH, similar response currents are obtained with the negatively charged ferricyanide and the uncharged mPMS (5.9 mu A and 6.7 mu A, respectively, for 10 mM glucose), however, no response current is obtained with hexaammineruthenium(III), which has a strong positive charge. These results show that access of even very small mediators with strong charges to a buried active center can be almost completely blocked by the protein.
  2017年11月16日, 研究論文（学術雑誌）, 共同, 17, 11, DOI(公開)(r-map), 2636
• Novel fungal FAD glucose dehydrogenase derived from Aspergillus niger for glucose enzyme sensor strips
  Koji Sode, Noya Loew, Yosuke Ohnishi, Hayato Tsuruta, Kazushige Mori, Katsuhiro Kojima, Wakako Tsugawa, Jeffrey T. LaBelle, David C. Klonoff
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  In this study, a novel fungus FAD dependent glucose dehydrogenase, derived from Aspergillus niger (AnGDH), was characterized. This enzyme's potential for the use as the enzyme for blood glucose monitor enzyme sensor strips was evaluated, especially by investigating the effect of the presence of xylose during glucose measurements. The substrate specificity of AnGDH towards glucose was investigated, and only xylose was found as a competing substrate. The specific catalytic efficiency for xylose compared to glucose was 1.8%. The specific activity of AnGDH for xylose at 5 mM concentration compared to glucose was 3.5%. No other sugars were used as substrate by this enzyme. The superior substrate specificity of AnGDH was also demonstrated in the performance of enzyme sensor strips. The impact of spiking xylose in a sample with physiological glucose concentrations on the sensor signals was investigated, and it was found that enzyme sensor strips using AnGDH were not affected at all by 5 mM (75 mg/dL) xylose. This is the first report of an enzyme sensor strip using a fungus derived FADGDH, which did not show any positive bias at a therapeutic level xylose concentration on the signal for a glucose sample. This clearly indicates the superiority of AnGDH over other conventionally used fungi derived FADGDHs in the application for SMBG sensor strips. The negligible activity of AnGDH towards xylose was also explained on the basis of a 3D structural model, which was compared to the 3D structures of A. flavus derived FADGDH and of two glucose oxidases.
  2017年08月18日, 研究論文（学術雑誌）, 共同, 87, DOI(公開)(r-map), 305, 311
• Electrochemical sensing system employing fructosamine 6-kinase enables glycated albumin measurement requiring no proteolytic digestion.
  Kameya M, Tsugawa W, Yamada-Tajima M, Hatada M, Suzuki K, Sakaguchi-Mikami A, Ferri S, Klonoff DC, Koji SODE
  Biotechnol J.
  2016年10月, 研究論文（学術雑誌）, 共同, 11, 6, DOI(公開)(r-map), 797, 804
• Continuous operation of an ultra-low-power microcontroller using glucose as the sole energy source,
  李仁榮、早出貴司、ノヤ・レーブ、津川若子、クリストファー･ロビン・ロウ、早出広司
  Biosens. Bioelectron
  2016年09月, 研究論文（学術雑誌）, 共同, 2016, 20116
• An Fe-S cluster in the conserved Cys-rich region in the catalytic subunit of FAD-dependentdehydrogenase complexes.
  doi: 10.1016/j.bioelechem.2016.01.010. Epub 2016 Feb 2.
  Bioelectrochemistry
  2016年01月, 研究論文（学術雑誌）, 共同, 2016, 112, DOI(公開)(r-map), 178, 83
• BioCapacitor: A novel principle for biosensors
  Koji Sode, Tomohiko Yamazaki, Inyoung Lee, Takuya Hanashi, Wakako Tsugawa
  BIOSENSORS & BIOELECTRONICS
  2015年08月07日, 共同, 76, DOI(公開)(r-map), 20, 28
• Engineering of dye-mediated dehydrogenase property of fructosyl amino acid oxidases by site-directed mutagenesis studies of its putative proton relay system.
  Kim S, Nibe E, Ferri S, Tsugawa W, Sode K.
  Biotechnol Lett.
  2010年08月, 研究論文（学術雑誌）, 共同, 32, 8, 1123, 9
• Wireless monotoring of blood glucose levels in flatfish with a needle biosensor
  Hideaki Endo,Eiji Takahashi, Masataka Murata, Hitoshi Ohnuki, Huifeng Ren, Wakako Tsugawa and Koji Sode
  Fisheries science
  2010年07月, 研究論文（学術雑誌）, 共同, 76, 4, 687, 694
• Motif-based search for a novel fructosyl peptide oxidase from genome databases.
  Kim S, Ferri S, Tsugawa W, Mori K, Sode K.
  Biotechnol Bioeng.
  2010年06月, 研究論文（学術雑誌）, 共同, 106, 3, 358, 66
• Enzyme fuel cell for cellulolytic sugara conversion employing FAD glucose dehydrogenase and carbon croth electrode on direct tlectron transfer principle.
  Desriani, Takuya Hanashi, Tomohiko Yamazaki, Tsugawa Wakako, Koji Sode
  The open electrochemistry journal.
  2010年02月, 研究論文（学術雑誌）, 共同, 2, 6, 10
• The construction of a glucose-sensing luciferase.
  種岡　坂口三上　山崎　津川　早出
  Biosens Bioelectron.
  2009年09月, 研究論文（学術雑誌）, 共同, 25, 1, 76, 81
• Review of fructosyl amino acid oxidase engineering research: a glimpse into the future of hemoglobin A1c biosensing.
  Ferri S, Kim S, Tsugawa W, Sode K.
  J Diabetes Sci Technol.
  2009年05月, 研究論文（学術雑誌）, 共同, 3, 3, 585, 92
• BioCapacitor--a novel category of biosensor.
  Hanashi T, Yamazaki T, Tsugawa W, Ferri S, Nakayama D, Tomiyama M, Ikebukuro K, Sode K.
  Biosens Bioelectron.
  2009年03月, 研究論文（学術雑誌）, 共同, 24, 7, 1837, 42
• Cumulative effect of amino acid substitution for development of fructosyl valine-specific fructosyl amine oxidase
  Seungsu Kim, Seiji Miura, Stefano Ferri, Wakako Tsugawa, Koji Sode
  Enzyme and microbial technology
  2009年01月, 研究論文（学術雑誌）, 共同, 44, 1, 52, 56
• Wireless enzyme sensor system for real-time monitoring of blood glucose levels in fish.
  Endo H, Yonemori Y, Hibi K, Ren H, Hayashi T, Tsugawa W, Sode K.
  Biosens Bioelectron.
  2009年05月, 研究論文（学術雑誌）, 共同, 24, 5, 1417, 1423
• A glimps into the future of hemoglobin A1c biosensing.
  Stefano Ferri, Seungsu Kim, Wakako Tsugawa, Koji Sode
  Journal of diabetes science and technology
  2009年01月, 研究論文（学術雑誌）, 共同, 3, 3, 585, 592
• Propionate sensor using coenzyme-A transferase and acyl-CoA oxidase.
  Sode K, Tsugawa W, Aoyagi M, Rajashekhara E, Watanabe K.
  Protein Pept Lett.
  2008年08月, 研究論文（学術雑誌）, 共同, 15, 8, 779, 81
• A Novel Wireless Glucose Sensor Employing Direct Electron Transfer Principle Based Enzyme Fuel Cell
  Noriko Kakehi, Tomohiko Yamazaki, Wakako Tsugawa and Koji Sode
  Biosensors & Bioelectronics
  2007年12月, 研究論文（学術雑誌）, 共同, 22-9-10, 2250, 5
• Development of Fructosyl Amine Oxidase Specific to Fructosyl Valine by Site-directed Mutagenesis
  Seiji Miura, Stefano Ferri, Wakako Tsugawa, Seungsu Kim and Koji Sode
  Protein Engineering, Design and Selection
  2007年08月, 研究論文（学術雑誌）, 共同, 21, 4, 233, 9
• Active Site Analysis of Fructosyl Amine Oxidase Using Homology Modeling and Site-directed Mutagenesis
  Seiji Miura, Stefano Ferri, Wakako Tsugawa, Seungsu Kim and Koji Sode
  Biotechnol.Lett
  2006年10月, 研究論文（学術雑誌）, 共同, 28, 23, 1895, 1900
• Novel Fluorescent Sensing System for α-Fructosyl Amino Acids Based on Engineered Fructosyl Amino Acid Binding Protein
  Akane Sakaguchi, Stefano Ferri, Wakako Tsugawa and Koji Sode
  Biosensors & Bioelectronics
  2006年09月, 研究論文（学術雑誌）, 共同, 2006, 9
• Isolation and Characterization of a Fructosyl-amine Oxidase from Arthrobacter sp FV1-1
  Stefano Ferri, Akane Sakaguchi, Hiroki Goto, Wakako Tsugawa, and Koji Sode
  Biotechnol.Lett
  2005年01月, 研究論文（学術雑誌）, 共同, 27, 27, 32
• Cloning and Expression of a Fructosyl-amine Oxidase from the Marine Yeast Pichia sp. N1-1
  Stefano Ferri, Seiji Miura, Akane Sakaguchi, Fumimasa Ishimura, Wakako Tsugawa and Koji Sode
  Marine Biotechnology
  2004年06月, 研究論文（学術雑誌）, 共同, 6, 625, 632
• Development of Highly-Sensitive Fructosyl-valine Enzyme Sensor Employing Recombinant Fructosyl Amine Oxidase
  Akane Sakaguchi, Wakako Tsugawa, Stefano Ferri and Koji Sode
  Electrochemistry
  2003年06月, 研究論文（学術雑誌）, 共同, 71, 6, 442, 445
• Amperometric Glucose Sensor Using Thermostable Co-factor Binding Glucose Dehydrogenase
  Yukie Nakazawa, Tomohiko Yamazaki, Wakako Tsugawa, Kazunori Ikebukuro and Koji Sode
  IEEJ Transactions on Sensors and Micromachines
  2003年06月, 研究論文（学術雑誌）, 共同, 123, 6, 185, 189
• X-ray structure of the direct electron transfer-type FAD glucose dehydrogenase catalytic subunit complexed with a small subunit
  Hiromi YOSHIDA , Katsuhiro KOJIMA , Wakako TSUGAWA , Shigehiro KAMITORI and Koji SODE
  Photon Factory Activity Report 2020
  2021年, （MISC）研究論文, 共同, 38
• Rapid, convenient, and highly sensitive detection of human hemoglobin in serum using a high-affinity bivalent antibody–enzyme complex
  Daimei Miura, Hayato Kimura, Wakako Tsugawa, Kazunori Ikebukuro, Koji Sode, Ryutaro Asano
  Talanta
  2021年11月01日, 研究論文（学術雑誌）, 共同, 234, 1, DOI(公開)(r-map), 122638
• Continuous electrochemical monitoring of L-glutamine using redox-probe-modified L-glutamine-binding protein based on intermittent pulse amperometry
  Shouhei Takamatsu, Jinhee Lee, Ryutaro Asano, Wakako Tsugawa, Kazunori Ikebukuro, Koji Sode
  Sensors and Actuators B: Chemical
  2021年11月01日, 研究論文（学術雑誌）, 共同, 346, 1, DOI(公開)(r-map), 346
• A self-powered glucose sensor based on BioCapacitor principle with micro-sized enzyme anode employing direct electron transfer type FADGDH
  Inyoung Lee,, Junko Okuda-Shimazaki, Wakako Tsugawa, Kazunori Ikebukuro, Koji Sode
  J. Phys. Energy
  2021年03月01日, 研究論文（学術雑誌）, 共同, 3, DOI(公開)(r-map), 034009
• Rapid and homogeneous electrochemical detection by fabricating a high affinity bispecific antibody-enzyme complex using two Catcher/Tag systems
  HayatoKimuraaDaimeiMiuraaWakakoTsugawaaKazunoriIkebukuroaKojiSodebRyutaroAsano
  Biosensors and Bioelectronics
  2021年03月01日, 研究論文（学術雑誌）, 共同, 175, 1, DOI(公開)(r-map), 112885
• Rational Design of Direct Electron Transfer Type L-lactate Dehydrogenase for the Development of Multiplexed Biosensor
  Kentaro Hiraka, WakakoTsugawa, Ryutaro Asano, Murat A.Yokus, Kazunori Ikebukuro, Michael A.Daniele, Koji Sode
  BIOSENSORS & BIOELECTRONICS
  2020年12月25日, 研究論文（学術雑誌）, 共同, DOI(公開)(r-map)
• Strategic design and improvement of the internal electron transfer of heme b domain-fused glucose dehydrogenase for use in direct electron transfer-type glucose sensors
  Kohei Ito, JunkoOkuda-Shimazaki , Katsuhiro Kojima, Kazushige Mori, Wakako Tsugawa, Ryutaro Asano, Kazunori Ikebukuro, KojiSode
  BIOSENSORS & BIOELECTRONICS
  2020年12月17日, 研究論文（学術雑誌）, 共同, DOI(公開)(r-map)
• Creation of a novel DET type FAD glucose dehydrogenase harboring Escherichia coli derived cytochrome b(562) as an electron transfer domain
  Yanase, Takumi; Okuda-Shimazaki, Junko; Mori, Kazushige; Kojima, Katsuhiro; Tsugawa, Wakako; Sode, Koji
  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
  ACADEMIC PRESS INC ELSEVIER SCIENCE
  Fungi-derived flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenases (FADGDHs) are the most popular and advanced enzymes for SMBG sensors because of their high substrate specificity toward glucose and oxygen insensitivity. However, this type of FADGDH hardly shows direct electron transfer (DET) ability. In this study, we developed a new DET-type FADGDH by harboring Cytochrome b(562) (cyt b(562)) derived from Escherichia coli as the electron transfer domain. The structural genes encoding fusion enzymes composed of cyt b(562) at either the N- or C-terminus of fungal FADGDH, (cyt b(562)-GDH or GDH-cyt b(562)), were constructed, recombinantly expressed, and characteristics of the fusion proteins were investigated. Both constructed fusion enzymes were successfully expressed in E. coli, as the soluble and GDH active proteins, showing cyt b(562) specific redox properties. Thusconstructed fusion proteins showed internal electron transfer between FAD in FADGDH and fused cyt b(562). Consequently, both cyt b(562)-GDH and GDH-cyt b(562) showed DET abilities toward electrode. Interestingly, cyt b(562)-GDH showed much rapid internal electron transfer and higher DET ability than GDH-cyt b(562). Thus, we demonstrated the construction and production of a new DET-type FADGDH using E.coli as the host cells, which is advantageous for future industrial application and further engineering. (C) 2020 Elsevier Inc. All rights reserved.
  2020年09月10日, 研究論文（学術雑誌）, 共同, 530, 1, 0006-291X, DOI(公開)(r-map), 82, 86
• Alteration of Electron Acceptor Preferences in the Oxidative Half-Reaction of Flavin-Dependent Oxidases and Dehydrogenases
  Hiraka, Kentaro; Tsugawa, Wakako; Sode, Koji
  INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
  MDPI
  In this review, recent progress in the engineering of the oxidative half-reaction of flavin-dependent oxidases and dehydrogenases is discussed, considering their current and future applications in bioelectrochemical studies, such as for the development of biosensors and biofuel cells. There have been two approaches in the studies of oxidative half-reaction: engineering of the oxidative half-reaction with oxygen, and engineering of the preference for artificial electron acceptors. The challenges for engineering oxidative half-reactions with oxygen are further categorized into the following approaches: (1) mutation to the putative residues that compose the cavity where oxygen may be located, (2) investigation of the vicinities where the reaction with oxygen may take place, and (3) investigation of possible oxygen access routes to the isoalloxazine ring. Among these approaches, introducing a mutation at the oxygen access route to the isoalloxazine ring represents the most versatile and effective strategy. Studies to engineer the preference of artificial electron acceptors are categorized into three different approaches: (1) engineering of the charge at the residues around the substrate entrance, (2) engineering of a cavity in the vicinity of flavin, and (3) decreasing the glycosylation degree of enzymes. Among these approaches, altering the charge in the vicinity where the electron acceptor may be accessed will be most relevant.
  2020年06月, 研究論文（学術雑誌）, 共同, 21, 11, DOI(公開)(r-map)
• Employment of 1-Methoxy-5-Ethyl Phenazinium Ethyl Sulfate as a Stable Electron Mediator in Flavin Oxidoreductases-Based Sensors
  Fitriana, Maya; Loew, Noya; Witarto, Arief Budi; Ikebukuro, Kazunori; Sode, Koji; Tsugawa, Wakako
  SENSORS
  MDPI
  In this paper, a novel electron mediator, 1-methoxy-5-ethyl phenazinium ethyl sulfate (mPES), was introduced as a versatile mediator for disposable enzyme sensor strips, employing representative flavin oxidoreductases, lactate oxidase (LOx), glucose dehydrogenase (GDH), and fructosyl peptide oxidase (FPOx). A disposable lactate enzyme sensor with oxygen insensitive Aerococcus viridans-derived engineered LOx (AvLOx), with A96L mutant as the enzyme, was constructed. The constructed lactate sensor exhibited a high sensitivity (0.73 +/- 0.12 mu A/mM) and wide linear range (0-50 mM lactate), showings that mPES functions as an effective mediator for AvLOx. Employing mPES as mediator allowed this amperometric lactate sensor to be operated at a relatively low potential of +0.2 V to 0 V vs. Ag/AgCl, thus avoiding interference from uric acid and acetaminophen. The lactate sensors were adequately stable for at least 48 days of storage at 25 degrees C. These results indicated that mPES can be replaced with 1-methoxy-5-methyl phenazinium methyl sulfate (mPMS), which we previously reported as the best mediator for AvLOx-based lactate sensors. Furthermore, this study revealed that mPES can be used as an effective electron mediator for the enzyme sensors employing representative flavin oxidoreductases, GDH-based glucose sensors, and FPOx-based hemoglobin A1c (HbA1c) sensors.
  2020年05月, 研究論文（学術雑誌）, 共同, 20, 10, DOI(公開)(r-map)
• Rational engineering of Aerococcus viridans L-lactate oxidase for the mediator modification to achieve quasi-direct electron transfer type lactate sensor
  Hiraka, Kentaro; Kojima, Katsuhiro; Tsugawa, Wakako; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji
  BIOSENSORS & BIOELECTRONICS
  ELSEVIER ADVANCED TECHNOLOGY
  The L-lactate oxidase (LOx) based lactate sensors are widely used for clinical diagnostics, sports medicine, and food quality control. However, dissolved oxygen interference and electroactive interferent effects are inherent issues of current lactate sensors. In this paper, a quasi-direct electron transfer (quasi-DET) type lactate sensor was developed using rationally engineered Aerococcus viridans LOx (AvLOx) modified with amine-reactive phenazine ethosulfate (PES). Since the modification of wild type AvLOx by PES did not result quasi-DET, engineered AvLOx with additional Lys residue was designed. The additional Lys residue was introduced by substituting residue locating on the surface of AvLOx, and within 20 angstrom of the isoalloxazine ring of FMN. Among several constructed mutants, Ala96Leu/Asn212Lys double mutant showed the highest dye-mediated dehydrogenase activity with negligible oxidase activity, showing quasi-DET properties after PES modification, when the enzyme was immobilized on screen printed carbon electrode. The constructed electrode did not show oxygen interference in cyclic voltammetric analysis and distinct catalytic current with 20 mM L-lactate. The sensor performance of a chronoamperometric L-lactate sensor employing PES modified Ala96Leu/Asn212Lys AvLOx, marked with linear range between 0 and 1 mM, with sensitivity of 13 mu A/mM.cm(2), and a limit of detection of 25 mu M for L-lactate. By applying -200 mV vs. Ag/AgCl, L-lactate could be monitored with negligible interference from 170 mu M ascorbic acid, 1.3 mM acetaminophen, 1.4 mM uric acid or 20 mM glucose. These results indicated that a quasi-DET type lactate sensor was developed that did not suffer from the interference of oxygen and representative electroactive ingredient compounds.
  2020年03月01日, 研究論文（学術雑誌）, 共同, 151, 0956-5663, DOI(公開)(r-map)
• Engineered Glucose Oxidase Capable of Quasi-Direct Electron Transfer after a Quick-and-Easy Modification with a Mediator
  Suzuki, Nanami; Lee, Jinhee; Loew, Noya; Takahashi-Inose, Yuka; Okuda-Shimazaki, Junko; Kojima, Katsuhiro; Mori, Kazushige; Tsugawa, Wakako; Sode, Koji
  INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
  MDPI
  Glucose oxidase (GOx) has been widely utilized for monitoring glycemic levels due to its availability, high activity, and specificity toward glucose. Among the three generations of electrochemical glucose sensor principles, direct electron transfer (DET)-based third-generation sensors are considered the ideal principle since the measurements can be carried out in the absence of a free redox mediator in the solution without the impact of oxygen and at a low enough potential for amperometric measurement to avoid the effect of electrochemically active interferences. However, natural GOx is not capable of DET. Therefore, a simple and rapid strategy to create DET-capable GOx is desired. In this study, we designed engineered GOx, which was made readily available for single-step modification with a redox mediator (phenazine ethosulfate, PES) on its surface via a lysine residue rationally introduced into the enzyme. Thus, PES-modified engineered GOx showed a quasi-DET response upon the addition of glucose. This strategy and the obtained results will contribute to the further development of quasi-DET GOx-based glucose monitoring dedicated to precise and accurate glycemic control for diabetic patient care.
  2020年02月, 研究論文（学術雑誌）, 共同, 21, 3, DOI(公開)(r-map)
• Engineered Glucose Oxidase Capable of Quasi-Direct Electron Transfer after a Quick-and-Easy Modification with a Mediator
  Nanami Suzuki, Jinhee Lee, Noya Loew, Yuka Takahashi-Inose, Junko Okuda-Shimazaki, Katsuhiro Kojima, Kazushige Mori, Wakako Tsugawa, Koji Sode
  INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
  2020年02月02日, 研究論文（学術雑誌）, 共同, 21, 3, DOI(公開)(r-map)
• Development of an Electrochemical Adenosine　Diphosphate (ADP) Sensing Principle Employing ADP-dependent Kinase
  Wakako Tsugawa, Hinako Futagami and Koji Sode
  Sensors and Materials
  2015年09月, 研究論文（学術雑誌）, 共同, 27, 9, 839–845
• Advancing the Development of Glycated Protein Biosensing Technology Next-Generation Sensing Molecules."
  Kameya, M., Sakaguchi-Mikami, A., Ferri, S., Tsugawa, W., & Sode, K.
  Journal of diabetes science and technology
  2015年09月, 研究論文（学術雑誌）, 共同, 9, 2, 183, 191
• The development of an autonomous self-powered bio-sensing actuator
  Takuya Hanashi,Tomohiko Yamazaki, Hiroshi Tanaka, Kazunori Ikebukuro, Wakako Tsugawa ,Koji Sode
  Sensors and Actuators B: Chemical
  2014年06月, 研究論文（学術雑誌）, 共同, 196, DOI(公開)(r-map), 429, 433
• Engineering glucose oxidase to minimize the influence of oxygen on sensor response
  Yohei Horaguchi, Shoko Saito, Katsuhiro Kojima,Wakako Tsugawa, Stefano Ferri,Koji Sode
  Electrochimica Acta
  2014年04月, 研究論文（学術雑誌）, 共同, 126, 20, DOI(公開)(r-map), 158, 161
• Identification and functional analysis of fructosyl amino acid-binding protein from Gram-positive bacterium Arthrobacter sp.
  A Sakaguchi-Mikami; S Ferri; S Katayama; Wakako Tsugawa; K Sode
  Journal of applied microbiology
  2013年05月, 研究論文（学術雑誌）, 共同, 114, 5, 1449, 1456
• Mutational analysis of the oxygen-binding site of cholesterol oxidase and its impact on dye-mediated dehydrogenase activity
  Katsuhiro Kojima,Tomohiko Kobayashi, Wakako Tsugawa, Stefano Ferri, Koji Sode
  Journal of Molecular Catalysis B: Enzymatic
  2013年04月, 研究論文（学術雑誌）, 共同, 88, DOI(公開)(r-map), 41, 46
• Cloning and characterization of fructosamine-6-kinase from Arthrobacter aurescens.
  Akane Sakaguchi-Mikami; Stefano Ferri; Miho Kameya; Wakako Tsugawa; Koji Sode
  Applied biochemistry and biotechnology
  2013年03月, 研究論文（学術雑誌）, 共同, 170, 3, 710, 717
• Engineering fructosyl peptide oxidase to improve activity toward the fructosyl hexapeptide standard for HbA1c measurement.
  Stefano Ferri; Yusuke Miyamoto; Akane Sakaguchi-Mikami; Wakako Tsugawa; Koji Sode
  Molecular biotechnology
  2013年03月, 研究論文（学術雑誌）, 共同, 54, 3, 939, 943
• Substrate specificity engineering of Escherichia coli derived fructosamine 6-kinase.
  Kojima K, Mikami-Sakaguchi A, Kameya M, Miyamoto Y, Ferri S, Tsugawa W, Sode K.
  Biotechnology Letters
  2013年02月, 研究論文（学術雑誌）, 共同, 35, 2, 253, 8
• 直接電子移動型酵素が拓くバイオエンジニアリング
  津川若子・早出広司
  電気化学会誌
  2013年01月, 共同, 81, 1, 20, 25
• Construction of mutant glucose oxidase with increased dye-mediated dehydrogenase activity.
  Yohei Horaguchi, Shoko Saito, Katsuhiro Kojima, Wakako Tsugawa, Stefano Ferri, Koji Sode
  Int J Mol Sci.
  2012年11月, 研究論文（学術雑誌）, 共同, 13, 11, 14149, 57
• BioLC-oscillator: a self -powered wireless glucose- sensing system with the glucose dependent resonance frequency
  Takuya Hanashi, Tomohiko Yamazaki, Wakako Tsugawa, Kazunori Ikebukuro, Koji Sode
  Electrochemistry
  2012年05月, 研究論文（学術雑誌）, 共同, 80, 5, 367, 370
• Glucose monitoring by direct electron transfer needle -type miniaturized electrode.
  Hitomi Shimizu, Wakako Tsugawa
  Electrochemistry
  2012年05月, 研究論文（学術雑誌）, 共同, 80, 5, 375
• Sensitive electrochemical ATP assay combined with enzymatic ATP amplification reaction
  Daisuke Okamura, Kunihiko Machida, Takeshi Kawahara, Katsuhiro Kojima, Wakako Tsugawa
  Electrochemistry
  2012年05月, 研究論文（学術雑誌）, 共同, 80, 5, 334, 336
• Nitrous oxide sensing using oxygen-insensitive direct-electron- transfer- type nitrous oxide reductase
  Wakako Tsugawa, Hitomi Shimizu, Masahiro Tatara, Yoshiyuki Ueno, Katsuhiro Kojima, Koji Sode
  Electrochemistry
  2012年05月, 研究論文（学術雑誌）, 共同, 80, 5, 371, 374
• ヘモグロビンA1c（HbA1c）バイオセンサー
  三上あかね　フェリ・ステファノ　津川若子　早出広司
  臨床化学
  2011年11月, 共同, 40, 4, 304, 311
• Construction of engineered fructosyl peptidyl oxidase for enzyme sensor applications under normal atmospheric conditions
  Kim S, Nibe E, Tsugawa W, Kojima K, Ferri S, Sode K.
  Biotechnol Lett.
  2011年11月, 研究論文（学術雑誌）, 共同, [Epub, ahead of, print]2011 Nov 4.
• BioRadioTransmitter: A Self-Powered Wireless Glucose-Sensing System
  Hanashi T, Yamazaki T, Tsugawa W, Ikebukuro K, Sode K.
  J Diabetes Sci Technol
  2011年09月, 研究論文（学術雑誌）, 共同, 5, 5, 1030, 5
• Relationship Between Serum N-Carbamoyl-β-D-Glucopyranosylamine Level and Renal Failure
  Tetsuro Hamafuji, Wakako Tsguawa and Koji Sode
  Renal Failure
  2003年01月, 研究論文（学術雑誌）, 共同, 25, 1, 115, 121
• Isolation and Identification of N-Carbamoyl-β-D-Glucopyranosylamine from Human Serum
  Testuro Hamafuji, Wakako Tsugawa and Koji Sode
  J.Biochem.Mol.Biol.Biophys
  2002年12月, 研究論文（学術雑誌）, 共同, 6, 5, 315, 318
• Development of a Flow Injection Analysis (FIA) Enzyme Sensor for Fructosyl Amine Monitoring
  Kinuko Ogawa, Daniela Stöllner, Frieder Scheller, Axel Warsinke, Fumimasa Ishimura, Wakako Tsugawa, Stefano Ferri and Koji Sode
  Analytical and Bioanalytical Chemistry
  2002年06月, 研究論文（学術雑誌）, 共同, 373, 211, 214
• Clinical Application of Serum 1,5-Anhydroglucitol Assay Method Using Glucose 3-Dehydrogenase
  Tetsuro Hamafuji, Wakako Tsugawa and Koji Sode
  J.Clinical Laboratory Analysis
  2002年06月, 研究論文（学術雑誌）, 共同, 16, 6, 299, 303
• Multi-sugar Analysis System Using Novel Glucose 3-Dehydrogenase Electrode
  Tetsuro Hamafuji, Yosuke Takano, Wakako Tsugawa and Koji Sode
  Instrumentation Science & Technology
  2002年01月, 研究論文（学術雑誌）, 共同, 30, 1, 97, 105
• Fructosyl Amine Sensing Based on Prussian Blue Modified Enzyme Electrode
  Wakako Tsugawa, Kinuko Ogawa, Fumimasa Ishimura and Koji Sode
  Electrochemistry
  2001年12月, 研究論文（学術雑誌）, 共同, 69, 12, 973, 975
• Enzymatic Synthesis of a Novel Trehalose Derivative, 3,3’-Diketotrehalose, and Its Potential Application as the Trehalase Enzyme Inhibitor
  Koji Sode, Eri Akaike, Hiroto Sugiura and Wakako Tsugawa
  FEBS Lett
  2001年06月, 研究論文（学術雑誌）, 共同, 489, 42, 45
• Cloning and Expression of Glucose 3-Dehydrogenase from Halomonas sp. α-15 in Escherichia coli
  Katsuhiro Kojima, Wakako Tsugawa and Koji Sode
  Biochem.Biophys. Res.Commun..
  2001年06月, 研究論文（学術雑誌）, 共同, 282, 21, 27
• A New Concept for the Construction of an Artificail Dehydrogenase for Fructosyl Amine Compounds and Its Application for an Amperometric Fructosyl Amine Sensor
  Koji Sode, Yuka Takahashi, Shigenori Ohta, Wakako Tsugawa and Tomohiko Yamazaki
  Anal.Chim.Acta
  2001年06月, 研究論文（学術雑誌）, 共同, 435, 151, 156
• Biodegradation of Formaldehyde by a Formaldehyde-Resistant Bacterium Isolated from Seawater
  Tomohiko Yamazaki,Tsugawa Wakako and Koji Sode
  Appl.Biochem.Biotechnol
  2001年06月, 研究論文（学術雑誌）, 共同, 91, 93, 213, 217
• Screening and Characterization of Fructosyl-valine Assimilating Marine Microorganisms
  Koji Sode, Fumimasa Ishimuda and Wakako Tsugawa
  Marine Biotechnol
  2001年03月, 研究論文（学術雑誌）, 共同, 3, 126, 132
• Development of an Enzyme Sensor Utilizing a Novel Fructosyl Amine Oxidase from a Marine Yeast
  Wakako Tsugawa, Fumimasa Ishimura, Kinuko Ogawa and Koji Sode
  Electrochemistry
  2000年11月, 研究論文（学術雑誌）, 共同, 68, 11, 869, 871
• Construction and Characterization of Glucose Enzyme Sensor Employing Engineered Water Soluble PQQ Glucose Dehydrogenase with Improved Thermal Stability
  Yuka Takahashi, Satoshi Igarashi, Yukie Nakazawa, Wakako Tsugawa and Koji Sode
  Electrochemistry
  2000年11月, 研究論文（学術雑誌）, 共同, 68, 11, 907, 911
• Improvement of Enatioselectivity of Chiral Organophosphate Insecticide Hydrolysis by Bacterial Phosphotriesterase
  Wakako Tsugawa, Hiroyuki Nakamura, Shokichi Ohuchi and Koji Sode
  Appl.Biochem.Biotechnol
  2000年06月, 研究論文（学術雑誌）, 共同, 84, 86, 311, 317
• Enzyme Electrochemical Preparation of a 3-Keto Derivative of 1,5-Anhydro-D-Glucitol Using Glucose-3-dehydrogenase
  Koji Sode, Hiroto Sugiura, Wakako Tsugawa, Yoshihumi Watazu and Tesuro Hamafuji
  Appl.Biochem.Biotechnol
  2000年06月, 研究論文（学術雑誌）, 共同, 84, 86, 947, 954
• Subunits Analyses of a Novel Thermostable Glucose Dehydrogenase Showing Different Temperature Properties According to Its Quaternary Structure
  Tomohiko Yamazaki, Wakako Tsugawa, and Koji Sode
  Appl.Biochm.Biotechnol
  1999年06月, 研究論文（学術雑誌）, 共同, 77-79, 325, 335
• Effect of Growth Substrates on Production of New Soluble Glucose 3-Dehydrogenase in Halomonas(Deleya )sp. α-15
  Katsuhiro Kojima, Wakako Tsugawa, Tetsuro Hamafuji, Yoshifumi Watazu and Koji Sode
  Appl.Biochem.Biotechnol
  1999年06月, 研究論文（学術雑誌）, 共同, 77-79, 827, 834
• Increased Thermal Stability of Glucose Dehydrogenase by Cross-linking Chemical Modification
  Tomohiko Yamazaki, Wakako Tsugawa and Koji Sode
  Biotechnol. Lett
  1999年06月, 研究論文（学術雑誌）, 共同, 21, 199, 202
• Fluorescent Measurement of 1,5-Anhydro-D-Glucitol Based on Novel Marine Bacterial Glucose Dehydrogenase
  Wakako Tsugawa, Nobuhiro Ogasawara and Koji Sode
  Enz.Microbiol.Biotechnol
  1998年06月, 研究論文（学術雑誌）, 共同, 22, 269, 274
• Alteration of Substrate Specifity of Galactose Oxidase by Chemical Cross-linking
  Koji Sode, Tomohiko Yamazaki, Tomonori Shimakita and Wakako Tsugawa
  Denki Kagaku
  1997年06月, 研究論文（学術雑誌）, 共同, 65, 6, 435, 439
• Growth Properties of a Marine Bacterium Capable of Assimilating 1,5-Anhydro-D-Glucitol as the Sole Carbon Source
  Wakako Tsugawa and Koji Sode
  J Mar Biotechnol
  1997年05月, 研究論文（学術雑誌）, 共同, 5, 201, 204
• Development of a Novel Conductometric Determination of Organophosphate Insecticides Using Phosphotriesterase
  Koji Sode, Hiromi Togo, Tomohiko Yamazaki, Wakako Tsugawa, Shookichi Oouchi
  Denki Kagaku
  1996年12月, 研究論文（学術雑誌）, 共同, 64, 12, 1234, 1238
• Purification of a Marine Bacterial Glucose Dehydrogenase from Cytophaga marinoflava and Its Application for Measurement of 1,5-Anhydro-D-Glucitol
  Wakako Tsugawa, Shuichi Horiuchi, Mitstuharu Tanaka, Hitoshi Wake and Koji Sode
  Appl. Biochem.Biotechnol
  1996年06月, 研究論文（学術雑誌）, 共同, 56, 301, 310
• A Novel Thermostable Glucose Dehydrogenase Varying Temperature Properties by Altering Its Quaternary Structure
  Koji Sode, Wakako Tsugawa, Tomohiko Yamazaki, Masato Watanabe, Nobuhiro Ogasawara, and Mitsuharu Tanaka
  Enz.Microb.Technol
  1996年06月, 研究論文（学術雑誌）, 共同, 19, 82, 85
• Effect of PQQ Glucose Dehydrogenase Overexpression in Escherichia coli on Sugar Dependent Respiration
  Koji Sode, Sayaka, Sugimoto, Mika Watanabe and Wakako Tsugawa
  J.Biotechnol
  1995年06月, 研究論文（学術雑誌）, 共同, 43, 41, 44
• Isolation of a Marine Bacterial Pyrroloquinoline Quinone-Dependent Glucose Dehydrogenase
  Koji Sode, Mitsuharu Tanaka, Kazunori Matsumura and Wakako Tsugawa
  J.Marine Biotechnol
  1995年02月, 研究論文（学術雑誌）, 共同, 2, 214, 218
• A Marine Bacterial Electrode for Sensitive Detection of 1,5-Anhydro-D-Glucitol
  Wakako Tsugawa and Koji Sode
  Denki Kagaku
  1994年12月, 研究論文（学術雑誌）, 共同, 63, 12, 1131, 1133
• Enzyme Sensor for Measurement of 1,5-Anhydro-D-Glucitol Utilizing a Novel Marine Bacterial Glucose Dehydrogenase
  Koji Sode, Wakako Tsugawa, Shizuko Kumazawa and Tadashi Matsunaga
  Proc. of IUMRS '93, Trans. Mat.Res. Soc. Jpn
  1994年06月, 研究論文（学術雑誌）, 共同, 15, 445, 447
• TRANSGLUTAMINASE IN ENDOTHELIAL CELLS FROM BOVINE CAROTID ARTERY
  SOICHI KOJIMA, HIROMI HAGIWARA, WAKAKO SOGA, MOTOYUKI SHIMONAKA, YUJI SAITO, YUJI INADA
  Biomedical Research
  1987年08月20日, 研究論文（学術雑誌）, 共同, 8, 1, DOI(公開)(r-map), 25, 29
• Visible fibrinolysis by endothelial cells: Effect of vitamins and sterols
  Soichi Kojima, Wakako Soga, Hiromi Hagiwara, Motoyuki Shimonaka, Yuji Saito & Yuji Inada
  Bioscience Reports
  springer
  1986年12月01日, 研究論文（学術雑誌）, 共同, 6, DOI(公開)(r-map), 1029, 1033
• Development of interdigitated electrode based enzyme sensor for triglyceride monitoring using engineered oxygen insensitive glycerol 3-phosphate oxidase
  Lee, Inyoung; Tong, Sheng; Hatada, Mika; Masterson, Adrianna; Hiraka, Kentaro; Okuda-Shimazaki, Junko; Tsugawa, Wakako; Hughes, Gary; Sode, Koji
  SENSORS AND ACTUATORS REPORTS
  ELSEVIER
  Triglycerides (TG) are an important biomarker of various diseases including hyper-lipidemia. Current emergent requirements for TG sensing include the development of electrochemical sensors for easy, cost-effective, disposable and single use triglyceride sensors. We propose a TG sensor using an engineered glycerol 3-phosphate oxidase (GlpO) and electrochemical detection. In this study, we aimed to develop an electrochemical TG sensor based on engineering the oxidative half reaction of GlpO to be oxygen insensitive, with the employment of an interdigitated electrode (IDE). To construct an oxygen insensitive mutant GlpO, we constructed a double mutant, Met104Phe/Asp169Glu, which revealed an engineered GlpO exhibiting repressed oxidase activity, while its dye-mediated dehydrogenase activity enhanced. The electrochemical enzyme sensor was constructed for the measurement of TG based on engineered GlpO using disposable screen-printed carbon electrode (SPCE) or disposable IDE using Ru complex as the electron mediator. By using the disposable IDE sensor, a highly sensitive electrochemical sensor for Glp was achieved, with the sensitivity of 0.4 µA/mM. The improved sensitivity was 2.4 times higher than that of SPCE (0.17 µA/mM), with LOD of 0.022 mM. Furthermore, the dry sensor strip, where the double mutant GlpO and Ru complex were dropcasted and dried on the IDE, was prepared and the Glp concentration was successfully measured in the delipidized serum solution. Therefore, by the employment of the established pretreatment method for TG to release Glp, a highly sensitive, accurate, easy and rapid electrochemical measurement method for TG will be developed using constructed M104F/D169E mutant GlpO with the combination of IDE.
  2023年06月, 研究論文（学術雑誌）, 共同, 5, 2666-0539, DOI(公開)(r-map), 359, 368
• Development of a Versatile Method to Construct Direct Electron Transfer-Type Enzyme Complexes Employing SpyCatcher/SpyTag System
  Yanase, Takumi; Okuda-Shimazaki, Junko; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji; Tsugawa, Wakako
  INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
  MDPI
  The electrochemical enzyme sensors based on direct electron transfer (DET)-type oxidoreductase-based enzymes are ideal for continuous and in vivo monitoring. However, the number and types of DET-type oxidoreductases are limited. The aim of this research is the development of a versatile method to create a DET-type oxidoreductase complex based on the SpyCatcher/SpyTag technique by preparing SpyCatcher-fused heme c and SpyTag-fused non-DET-type oxidoreductases, and by the in vitro formation of DET-type oxidoreductase complexes. A heme c containing an electron transfer protein derived from Rhizobium radiobacter (CYTc) was selected to prepare SpyCatcher-fused heme c. Three non-DET-type oxidoreductases were selected as candidates for the SpyTag-fused enzyme: fungi-derived flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase (GDH), an engineered FAD-dependent d-amino acid oxidase (DAAOx), and an engineered FMN-dependent l-lactate oxidase (LOx). CYTc-SpyCatcher (CYTc-SC) and SpyTag-Enzymes (ST-GDH, ST-DAAOx, ST-LOx) were prepared as soluble molecules while maintaining their redox properties and catalytic activities, respectively. CYTc-SC/ST-Enzyme complexes were formed by mixing CYTc-SpyCatcher and SpyTag-Enzymes, and the complexes retained their original enzymatic activity. Remarkably, the heme domain served as an electron acceptor from complexed enzymes by intramolecular electron transfer; consequently, all constructed CYTc-SC/ST-Enzyme complexes showed DET ability to the electrode, demonstrating the versatility of this method.
  2023年02月, 研究論文（学術雑誌）, 共同, 24, 3, DOI(公開)(r-map)

研究発表、招待講演等

• 直接電子移動型酵素の汎用的な構築法
  第33回日本MRS年次大会 (2023)
  2023年11月15日, 口頭発表（招待・特別）
• Development of an Electrochemical Biosensor of Ubiquitin C-Terminal Hydrolase L1 (UCH-L1) for the Assessment of Traumatic Brain Injury (TBI)
  ECS - The Electrochemical Society
  2020年12月01日, 口頭発表（一般）
• The Continuous 3 Month Operation of Open Circuit Potential Based Glucose Sensor Employing Direct Electron Transfer Type Fad Dependent Glucose Dehydrogenase
  The Electrochemical Society, ECS
  2020年12月01日, 口頭発表（一般）
• グリセロール3-リン酸酸化酵素の脱⽔素酵素化
  第14回バイオ関連化学シンポジウム
  2020年09月07日, ポスター発表
• Development of highly sensitive anti-human hemoglobin sensing elements using a universal fabrication technology for antibody-enzyme complex
  第20回日本蛋白質科学会年会
  2020年07月07日, ポスター発表
• イヌの乳腺腫瘍のアミロイド沈着より見出だされたα-S1-カゼインの凝集特性評価（東
  日本化学会第100春季年会(2020)
  2020年03月22日, 口頭発表（一般）
• がん細胞とNK細胞間を架橋する低分子二重特異性抗体の機能的な分子構造
  日本化学会第100春季年会(2020)
  2020年03月22日, 口頭発表（一般）
• グルタミン結合タンパク質を用いる電気化学的in situ,リアルタイム・連続グルタミン計測システムの開発
  電気化学会第87回大会
  2020年03月17日, 口頭発表（一般）
• 電極表面配向制御による真菌由来FAD依存型グルコース脱水素酵素の直接電子移動能の検証
  電気化学会第87回大会
  2020年03月17日, 口頭発表（一般）
• 谷中
  酵素工学研究会
  2019年11月11日, ポスター発表
• 乳酸
  酵素工学研究会
  2019年11月11日, ポスター発表
• G3DH
  酵素工学研究会
  2019年11月11日, ポスター発表
• ケカビ由来新規FAD依存型グルコース脱水素酵素の組換え生産
  酵素工学研究会
  2019年11月11日, ポスター発表
• 改良型フラボシトクロムb2を用いた直接電子移動型乳酸センサーの開発
  2019年電気化学秋季大会
  2019年09月06日, 口頭発表（一般）
• Employing a stable electron mediator for disposable enzyme sensor
  2019年電気化学秋季大会
  2019年09月06日, 口頭発表（一般）
• フローインジェクション分析法による改良型フルクトシルペプチド酸化酵素を用いたHbA1cセンサーの構築
  2019年電気化学秋季大会
  2019年09月05日, 口頭発表（一般）
• 新規電気化学イムノセンサーへの展開を目指した、メディエーター修飾ペプチドの応用
  2019年電気化学秋季大会
  2019年09月05日, 口頭発表（一般）
• cyt-GDH
  日本化学会
  2019年07月, 口頭発表（一般）
• 直接電子移動型グルコース脱水素酵素を用いた第三世代インピーダンスセンサーの検討
  2019年 電気化学会第86回大会
  2019年03月29日, 口頭発表（一般）
• HbA1cのFIA計測を目指した、脱水素酵素化二重変異体 フルクトシルペプチド酸化酵素による擬直接電子移動型センサーの構築
  2019年 電気化学会第86回大会
  2019年03月29日, 口頭発表（一般）
• 抗体酵素複合体の汎用的な作製技術を応用した電気化学的ヘモグロビンセンシング素子の開発
  2019年 電気化学会第86回大会
  2019年03月28日, 口頭発表（一般）
• arPES修飾insulin計測
  MRS-J
  2018年12月19日, ポスター発表
• 直接電子移動型酵素を用いた開回路型グルコースセンサー
  MRS-J
  2018年12月19日, 口頭発表（招待・特別）
• Biosensor for healthcare
  2nd international conference on
  2018年12月01日, 口頭発表（招待・特別）
• Development of an amperometric lactate sensor utilizing Aerococcus viridans lactate oxidase
  第78回酵素工学研究会
  2017年10月06日, ポスター発表
• フルクトサミン-6-キナーゼを用いた新規糖化アルブミン計測法の開発
  第78回酵素工学研究会
  2017年10月06日, ポスター発表
• グルコシド3-脱水素酵素における電子伝達経路の解明
  第78回酵素工学研究会
  2017年10月06日, ポスター発表
• 乳酸酸化酵素における酸素のアクセス経路解析
  第78回酵素工学研究会
  2017年10月06日, ポスター発表
• 電子伝達サブユニットの改良とFADグルコース脱水素酵素複合体の直接電子移動能の改変
  2017年電気化学秋季大会
  2017年09月11日, 口頭発表（一般）
• 直接電子移動型グルコースセンサーにおける酵素固定化自己組織化単分子膜の最適化
  2017年電気化学秋季大会
  2017年09月10日, 口頭発表（一般）
• グルコシド3-脱水素酵素の分子内および分子間電子伝達経路の解明
  2017年電気化学秋季大会
  2017年09月10日, 口頭発表（一般）
• 酵素電気化学インピーダンス計測法に基づく新規バイオセ ンシング技術の開発
  2017年電気化学秋季大会
  2017年09月10日, 口頭発表（一般）
• 真菌由来FAD依存型グルコース酸化還元酵素の人工電子受容体選択性の改良
  2017年電気化学秋季大会
  2017年09月10日, 口頭発表（一般）
• 脱水素酵素化した改良型乳酸酸化酵素の構築と乳酸センサーへの応用
  2017年電気化学秋季大会
  2017年09月10日, 口頭発表（一般）
• Electrochemical Properties of Glucoside 3-Dehydrogenase, an Enzyme for 1,5-Anhydro-D-Glucitol Sensing
  BES 2017
  2017年07月07日, 口頭発表（一般）
• Direct Electron Transfer Type Glucose Sensor with Self-Assembled Monolayer (SAM) Immobilized FAD Glucose Dehydrogenase Complex
  BES 2017
  2017年07月04日, 口頭発表（一般）
• Development of 2.5th Generation Biosensors
  BES 2017
  2017年07月04日, 口頭発表（一般）
• Interdigitated Array Microelectrodes as Novel Platform Technology for Disposable Enzyme Sensor Strips
  BES 2017
  2017年07月04日, 口頭発表（一般）
• Analyses of the electron transfer pathway of direct electron transfer type iron sulfur flavo cytochrome type glucose dehydrogenase complex ~enzyme for continuous glucose monitoring
  BES 2017
  2017年07月04日, 口頭発表（一般）
• L-Lactate biosensor employing engineered lactate oxidase with minimized oxygen interference
  5th international conference on Bio-sensing technology
  2017年05月07日, ポスター発表
• 鉄硫黄フラボシトクロム型グルコース脱水素酵素複合体の分子間・分子内電子伝達経路の検討
  電気化学会第84回大会
  2017年03月26日, 口頭発表（一般）
• ホームメイドディスポーザブル金電極を用いた電気化学インピーダンス法によるα-シヌクレインの解析
  電気化学会第84回大会
  2017年03月26日, 口頭発表（一般）
• 電気化学センサーへの応用を目指したグルコース脱水素酵素融合抗EGFR抗体の作製と機能評価
  電気化学会第84回大会
  2017年03月25日, 口頭発表（一般）
• Aspergillus flavus由来FAD依存型グルコース脱水素酵素変異体における基質特異性向上機序の解析
  電気化学会第84回大会
  2017年03月25日, 口頭発表（一般）
• 新規糖化アルブミン計測用センサへの応用を目指したフルクトサミン6?キナーゼの機能評価
  電気化学会第84回大会
  2017年03月25日, 口頭発表（一般）
• 自己組織化単分子膜(SAM)により固定化された直接電子移動型FAD依存型グルコース脱水素酵素の電気化学的挙動の解析
  電気化学会第84回大会
  2017年03月25日, 口頭発表（一般）
• Escherichia coli由来フルクトサミン6-キナーゼの構造解析
  日本化学会第97春季年会
  2017年03月17日, 口頭発表（一般）
• Rhizobium radiobacter由来グルコシド3-脱水素酵素におけるCys繰り返し配列の機能解析
  日本化学会第97春季年会
  2017年03月17日, 口頭発表（一般）
• Structural Based Investigation of Artificial Electron Acceptor Interaction of Fungus Derived Flavin Adenine Dinucleotide Dependent Glucose Dehydrogenase .
  Prime 2016
  2016年10月07日, その他
• Investigation of Intra- or Inter-Electron Transfer Pathway of Multiheme Electron Transfer Subunit of Iron Sulfur Flavo Cytochrome Type Glucose Dehydrogenase Complex
  Prime 2016
  2016年10月07日, 口頭発表（一般）
• Electrochemical Measurement of Coenzyme Q10 Using Disposable Interdigitatad Array Electrode .
  Prime 2016
  2016年10月07日, 口頭発表（一般）
• Disposable interdigitated array electrode as an innovative platform technology for the disposable amperometric enzyme sensors.
  Biosensors 2016
  2016年05月25日, ポスター発表
• Disposable interdigitated array electrode employing high sensitive enzyme sensor for glycated albumin measurement
  Biosensors 2016
  2016年05月25日, 口頭発表（一般）
• A self-powered, stand alone, autonomous, wireless real time glucose sensing system
  Biosensors 2016
  2016年05月25日, 口頭発表（一般）
• くし形電極を用いたコエンザイムQ10の電気化学的計測
  電気化学会第83回大会
  2016年03月31日, 口頭発表（一般）
• くし型電極を用いる使い捨て型グルコースセンサーの開発
  電気化学会第83回大会
  2016年03月30日, 口頭発表（一般）
• FAD結合型グルコース脱水素酵素複合体の電子伝達サブユニットへの変異導入とその電気化学的評価
  電気化学会第83回大会
  2016年03月30日, 口頭発表（一般）
• 使い捨て型電極を用いる糖化アルブミン計測用高感度酵素センサの開発
  電気化学秋季大会
  2015年09月12日, 口頭発表（一般）
• 微小電極を用いるバイオキャパシタ型グルコース計測システムの開発
  電気化学秋季大会
  2015年09月12日, 口頭発表（一般）
• FAD結合型グルコース脱水素酵素複合体の電子伝達サブユニットへの変異導入とその特性検討
  電気化学秋季大会
  2015年09月12日, 口頭発表（一般）
• ADを補酵素とするグルコース脱水素酵素複合体の触媒ドメインにおける鉄硫黄クラスタの同定（東京農工大学,物質・材料研究機構,(有)アルティザイム・インターナショナル）
  2015年電気化学秋季大会
  2015年09月12日, 口頭発表（一般）
• グルコース酸化酵素とFADを補酵素とするグルコース脱水素酵素のキメラ酵素の構築
  日本化学会第95春季年会(2015)
  2015年03月27日, 口頭発表（一般）
• グルコース脱水素酵素複合体の触媒ドメインにおけるシステインクラスタの機能解析
  日本化学会第95春季年会(2015)
  2015年03月27日, 口頭発表（一般）
• ｸﾞﾙｺｰｽ脱水素酵素複合体の電子伝達サブユニットへの変異導入とその特性検討
  日本化学会第95春季年会(2015)
  2015年03月27日, 口頭発表（一般）
• 新奇酸化還元酵素を応用した電気化学バイオセンシング技術
  電気化学会第82回大会
  2015年03月18日, 口頭発表（招待・特別）
• 改変型FADGDHを用いる連続グルコース計測システムの開発
  電気化学会第82回大会
  2015年03月18日, 口頭発表（一般）
• バイオキャパシタ型連続グルコース計測システムの開発
  電気化学会第82回大会
  2015年03月18日, 口頭発表（一般）
• オスミウム修飾ポリマーを用いる連続計測用乳酸センサの開発
  電気化学会第82回大会
  2015年03月18日, 口頭発表（一般）
• FADを補酵素とする直接電子移動型グルコース脱水素酵素の酸化還元特性の解析（
  電気化学会第82回大会
  2015年03月18日, 口頭発表（一般）
• 櫛型電極を用いた電気化学的なトロンビン活性の検出法の開発
  電気化学会第82回大会
  2015年03月15日, 口頭発表（一般）
• スクリーン印刷電極を用いる糖化アルブミン計測用酵素センサの開発
  電気化学会第82回大会
  2015年03月15日, 口頭発表（一般）
• 使い捨て型電極を用いるコエンザイムQの電気化学的計測
  電気化学会第82回大会
  2015年03月15日, 口頭発表（一般）
• Fructosyl amino acid catabolic enzymes / proteins in Gram positive bacteria, Arthrobacter species
  第13回日中韓酵素工学会議
  2014年11月, その他
• Development of direct electron transfer type enzyme sensors based on membrane bound dehydrogenases from acetic acid bacteria
  第13回日中韓酵素工学会議
  2014年11月, その他
• 酢酸菌由来膜結合性脱水素酵素を用いる直接電子移動型酵素電極の構築と特性検討
  CSJ化学フェスタ
  2014年10月, ポスター発表
• FADを補酵素とする直接電子移動型脱水素酵素複合体における触媒ドメインの機能解析
  2014年電気化学秋季大会
  2014年09月, 口頭発表（一般）
• バイオプロセスin situモニタリング用シングルユース型グルコースセンサの開発
  2014年電気化学秋季大会
  2014年09月, 口頭発表（一般）
• 直接電子移動型グルコースセンサーの皮下埋め込み型in vivo連続グルコース計測への応用
  2014年電気化学秋季大会
  2014年09月, 口頭発表（一般）
• Direct electron transfer enzyme sensor for in situ monitoring of mammalian cell culture bioprocesses
  Biosensors 2014
  2014年05月, ポスター発表
• Development of enzymatic formate sensing system for on-site evaluation ofbiodiesel fuel degradation
  Biosensors 2014
  2014年05月, ポスター発表
• First human in vivo trial of continuous glucose monitoring based on direct electron transfer principle
  Biosensors 2014
  2014年05月30日, 口頭発表（招待・特別）
• 酸化的半反応に関与する残基への変異導入によるグルコース酸化酵素の改良
  日本化学会第94春季年会(2014)
  2014年03月, 口頭発表（一般）
• 酢酸菌由来乳酸脱水素酵素の探索と特性検討
  日本化学会第94春季年会(2014)
  2014年03月, 口頭発表（一般）
• GMC酸化還元酵素ファミリー間での構造比較に基づくコレステロール酸化酵素の安定化
  日本化学会第94春季年会(2014)
  2014年03月, 口頭発表（一般）
• バイオプロセスリアルタイムin situモニタリング用直接電子移動型酵素センサーの開発
  日本化学会第94春季年会(2014)
  2014年03月, 口頭発表（一般）
• 直接電子移動型グルコースセンサーのin vivo持続血糖測定への応用
  電気化学会第81回大会
  2014年03月, 口頭発表（一般）
• 直接電子移動型グルコースセンサーを用いた浮遊性動物細胞培養におけるin situモニタリング
  電気化学会第81回大会
  2014年03月, 口頭発表（一般）
• αシヌクレイン超高次構造形成の シヌクレイン超高次構造形成の AC インピーダス法による解析 イ
  酵素工学研究会第70回講演会
  2013年10月25日, ポスター発表
• 酢酸菌由来乳酸脱水素酵素の特性検討
  酵素工学研究会第７０回講演会
  2013年10月25日, ポスター発表
• 直接電子移動型酵素センサーを用いた大腸菌培養のin situ モニタリング
  酵素工学研究会第70回講演会
  2013年10月25日, ポスター発表
• 交流インピーダンス法を用いた超高次構造を有するαシヌクレインの検出および測定
  日本化学会秋季事業 第３回 CSJフェスタ 2013
  2013年10月22日, ポスター発表
• NADH脱水素酵素の組み換え生産および特性評価
  日本化学会秋季事業 第３回 CSJフェスタ 2013
  2013年10月22日, ポスター発表
• 直接電子移動型グルコースセンサーを用いる動物細胞培養液のin situモニタリング
  2013年電気化学秋季大会
  2013年09月27日, その他
• 糖化アルブミン計測用使い捨て型酵素センサチップの開発
  2013年電気化学秋季大会
  2013年09月27日, その他
• 溶存酸素の影響を受けない改良型グルコース酸化酵素の開発
  2013年電気化学秋季大会
  2013年09月27日, その他
• バイオディーゼル燃料劣化のオンサイト評価用有機酸酵素分析システムの開発
  2013年電気化学秋季大会
  2013年09月27日, その他
• Glucose sensing employing direct electron transfer principle
  Enzyme Engineering XXII
  2013年09月26日, ポスター発表
• 直接電子移動型酵素センサーによるバイオプロセスのin situモニタリング
  公益社団法人電気化学会創立第80周年記念大会
  2013年03月29日, その他
• 使い捨て型電極を用いた交流インピーダンス法によるαシヌクレインの検出
  公益社団法人電気化学会創立第80周年記念大会
  2013年03月29日, その他
• バイオディーゼル燃料劣化のオンサイト評価用有機酸センサの開発
  公益社団法人電気化学会創立第80周年記念大会
  2013年03月29日, その他
• カビ由来新規FADグルコース脱水素酵素の基質特異性検討とグルコースセンサへの応用
  公益社団法人電気化学会創立第80周年記念大会
  2013年03月29日, その他
• 次世代型皮下連続式グルコース測定装置の開発　～直接電子移動型酵素が拓く新しいバイオセンシング～
  公益社団法人電気化学会創立第80周年記念大会
  2013年03月29日, 口頭発表（招待・特別）
• コレステロール酸化酵素の脱水素酵素化戦略
  日本農芸化学会　2013年度大会
  2013年03月26日, 口頭発表（一般）
• グルコース酸化酵素の脱水素酵素化戦略
  日本農芸化学会　2013年度大会
  2013年03月26日, 口頭発表（一般）
• ジスルフィド結合導入による真菌由来FADグルコース脱水素酵素の安定性改良
  日本農芸化学会　2013年度大会
  2013年03月26日, 口頭発表（一般）
• 糖化アミノ酸を基質とするクロストリジウム属由来酵素の解析
  日本化学会第93春季年会
  2013年03月22日, 口頭発表（一般）
• Engineering fructosyl peptide oxidase to improve activity toward the fructosyl hexapeptide standard for HbA1c measurement
  第22回　日本メイラード学会年会
  2012年12月21日, その他
• 新規フルクトサミン 6-キナーゼのゲノム情報に基づく検索
  第22回　日本メイラード学会年会
  2012年12月21日, ポスター発表
• フルクトサミン 6-キナーゼ、デグリカーゼからなる糖化蛋白質代謝系酵素の検索
  第22回　日本メイラード学会年会
  2012年12月21日, その他
• 新規フルクトサミン6-キナーゼのゲノム情報に基づく検索
  第64回日本生物工学会大会
  2012年10月24日, 口頭発表（一般）
• 新規コレステロール酸化酵素のゲノムデータベースに基づいた検索および特性評価
  第64回日本生物工学会大会
  2012年10月24日, 口頭発表（一般）
• Engineering of Catalytic Domain of Cellobiose Dehydrogenase and Its Application for the Direct Electron Transfer Type Enzyme Electrode
  【PRiME2012】PACIFIC RIM MEETING ON ELECTROCHEMICAL AND SOLID-STATE SCIENCE 2012
  2012年10月08日, その他
• Turning Oxidase into Dehydrogenase for Application to the Electrochemical Measurement
  【PRiME2012】PACIFIC RIM MEETING ON ELECTROCHEMICAL AND SOLID-STATE SCIENCE 2012
  2012年10月08日, その他
• Second Generation Continuous Glucose Sensing System Employing Direct Electron Transfer Principle
  【PRiME2012】PACIFIC RIM MEETING ON ELECTROCHEMICAL AND SOLID-STATE SCIENCE 2012
  2012年10月08日, その他
• Engineering Fungi Derived FAD Glucose Dehydrogenase and Its Application for Glucose Sensor Strip Employing Screen Printed Carbon Electrode
  【PRiME2012】PACIFIC RIM MEETING ON ELECTROCHEMICAL AND SOLID-STATE SCIENCE 2012
  2012年10月08日, その他
• Turning Glucose Oxidase into Essentially Dehydrogenase
  【PRiME2012】PACIFIC RIM MEETING ON ELECTROCHEMICAL AND SOLID-STATE SCIENCE 2012
  2012年10月08日, ポスター発表
• Altering glucose oxidase electron acceptor preference from oxygen to artificial electron acceptors
  The 10th Korea-China-Japan Joint Symposium on Enzyme Engineering
  2012年05月29日, 口頭発表（一般）
• Engineering of catalytic domain of cellobiose dehydrogenase from Phanerochaete chrysosporium
  The 10th Korea-China-Japan Joint Symposium on Enzyme Engineering
  2012年05月29日, 口頭発表（一般）
• Engineering glucose oxidase: Turning glucose oxidase into glucose dehydrogenase
  Biosensors 2012
  2012年05月16日, ポスター発表
• In vivo study of continuous glucose monitoring system employing direct electron transfer principle
  Biosensors 2012
  2012年05月16日, ポスター発表
• グルコース酸化酵素の酸素に対する反応性の改変
  日本化学会第92春季年会
  2012年03月, 口頭発表（一般）
• In vivo 計測を目指した、直接電子移動型グルコース脱水素酵素によるグルコース連続測定
  電気化学会第79回大会
  2012年03月31日, その他
• コレステロール酸化酵素の酸素に対する反応性の改変
  電気化学会第79回大会
  2012年03月31日, その他
• FADグルコース脱水素酵素を用いた直接電子移動型グルコースセンサーの長期連続運転
  日本化学会第92春季年会
  2012年03月28日, その他
• コレステロール酸化酵素のコレステロール脱水素酵素への変換
  日本化学会第92春季年会
  2012年03月28日, その他
• モチーフ解析に基づく新規フルクトシルペプチド酸化酵素の探索
  日本化学会第92春季年会
  2012年03月26日, その他
• 亜酸化窒素還元酵素をカソードに用いた酵素燃料電池の提案
  日本生物工学会63回大会
  2011年09月28日, その他
• 直接電子移動能を有するFAD 結合型脱水素酵素の組み換え生産
  日本生物工学会63回大会
  2011年09月26日, その他
• EcoTanker - 光アクチュエーターで操縦する大腸菌• マイクロ• タンカー
  日本生物工学会63回大会
  2011年09月26日, その他
• フルクトサミン-6-リン酸化酵素を用いた糖化蛋白質電気化学測定法の開発
  2011年電気化学秋季大会
  2011年09月11日, その他
• 架橋化グルコース脱水素酵素を用いた直接電子移動型グルコースセンサー
  2011年電気化学秋季大会
  2011年09月11日, その他
• グルコース脱水素酵素の基質特異性に依存した使い捨て型センサー応答の解析
  2011年電気化学秋季大会
  2011年09月10日, その他
• α-シヌクレインオリゴマーの電気化学的検出法の開発
  2011年電気化学秋季大会
  2011年09月10日, その他
• Novel Enzyme Sensor for Glycated Protein Biosensing Employing Fructosamine 6-Kinase
  ISE
  2011年09月06日, ポスター発表
• Electrochemical detection of oligomeric status of amyloid forming protein, alpha-synuclein
  ISE
  2011年09月06日, その他
• Direct electron transfer type continuous glucose monitoring system employing cross-linked FAD glucose dehydrogenase
  ISE
  2011年09月06日, その他
• Construction of glucose sensor strip employing engineered PQQGDH with minimized maltose effect
  ISE
  2011年09月06日, その他
• 複合基質を用いる自立型センシングアクチュエータの開発
  電気化学会第78回大会
  2011年03月31日, その他
• α-シヌクレイン凝集線維化の電気化学計測
  電気化学会第78回大会
  2011年03月30日, その他
• 基質特異性が異なる改変型酵素を用いる使い捨て型酵素センサの挙動解析
  電気化学会第78回大会
  2011年03月29日, その他
• 改良型糖化ペプチド酸化酵素を用いる酵素センサー
  電気化学会第78回大会
  2011年03月29日, その他
• 架橋化グルコース脱水素酵素を用いる直接電子移動型グルコースセンサーの連続計測における安定性の評価
  電気化学会第78回大会
  2011年03月28日, その他
• フルクトシルアミノ酸/ペプチド脱水素酵素の構築および糖化蛋白質センサーへの応用
  第20回　メイラード学会
  2010年09月17日, その他
• バイオキャパシタを用いた自立型センシングアクチュエータの開発
  2010年電気化学秋季大会
  2010年09月03日, 口頭発表（一般）
• ２酵素反応系を用いる中性脂肪計測用使い捨て型酵素センサの開発
  2010年電気化学秋季大会
  2010年09月02日, 口頭発表（一般）
• 直接電子移動能力を有するグルコース脱水素酵素の安定化
  2010年電気化学秋季大会
  2010年09月02日, 口頭発表（一般）
• 基質特異性が改変されたPQQグルコース脱水素酵素のSMBG用酵素としての評価
  2010年電気化学秋季大会
  2010年09月02日, 口頭発表（一般）
• 糖化ペプチド酸化酵素の機能改変による脱水素酵素化
  2010年電気化学秋季大会
  2010年09月02日, 口頭発表（一般）
• Bio-LC-Oscillator ~ a novel stand alone, wireless, self powered glucose sensing system ~
  Biosensors 2010
  2010年05月28日, ポスター発表
• The development of an autonomous self powered bio-sensing actuator
  Biosensors 2010
  2010年05月28日, 口頭発表（一般）
• Tuning oxidase into “dehydrogenase” ~ application of engineered fructosyl peptide oxidase for electrochemical glycated hemoglobin biosensing ~
  Biosensors 2010
  2010年05月28日, 口頭発表（一般）
• バイオLCオシレータを用いた自立型グルコースセンシングシステムの開発
  電気化学会第77回大会
  2010年03月31日, 口頭発表（一般）
• FADグルコース脱水素酵素を用いた直接電子移動型連続計測型グルコースセンサーの開発
  電気化学会第77回大会
  2010年03月31日, 口頭発表（一般）
• フルクトシルアミノ酸酸化酵素の電子受容体特異性の改変
  電気化学会第77回大会
  2010年03月30日, 口頭発表（一般）
• 変異導入によるフルクトシルアミノ酸酸化酵素における電子受容体反応性に関与する残基の解析
  日本化学会
  2010年03月30日, 口頭発表（一般）
• FAD結合型グルコース脱水素酵素の電子伝達サブユニットへの変異導入
  電気化学会第77回大会
  2010年03月29日, 口頭発表（一般）
• 魚類の血糖値測定ための機能性ポリマーを用いたワイヤレスグルコースセンサの試作
  電気化学会第77回大会
  2010年03月29日, 口頭発表（一般）
• フルクトシルアミノ酸酸化酵素の構造機能相関解析に基づく基質特異性を支配する領域の導出
  日本化学会
  2010年03月27日, 口頭発表（一般）
• 組換えハプトグロビンサブユニットのApoA-I認識サブユニットの解析とコレステロールエステラーゼ機能改良への応用
  生物工学会
  2009年09月24日, 口頭発表（一般）
• 亜酸化窒素のオンサイトモニタリング用酵素測定法の開発
  生物工学会
  2009年09月24日, 口頭発表（一般）
• バイオキャパシタを用いた自立型グルコースセンシングシステムの開発
  電気化学会大会
  2009年09月10日, 口頭発表（一般）
• 亜酸化窒素還元酵素を用いた温室効果ガスバイオセンシングシステムの開発
  日本電気化学会
  2009年09月10日, 口頭発表（一般）
• Novel Enzyme Sensor for Glycated Protein Biosensing Without the Proteolytic Processes
  ECS
  2009年05月28日, 口頭発表（一般）
• Development of Nitrous Oxide Enzyme Sensor Based on Direct Electron Transfer
  ECS
  2009年05月26日, 口頭発表（一般）
• 亜酸化窒素バイオセンサの開発
  電気化学会大会
  2009年03月31日, 口頭発表（一般）
• 亜酸化窒素バイオセンシングシステムの開発
  農芸化学会
  2009年03月28日, ポスター発表
• フルクトサミンリン酸化酵素を用いた糖化蛋白質測定用チップ型バイオセンサーの開発
  第18回　メイラード学会
  2008年11月29日, 口頭発表（一般）
• Cutting edge of glycated protein biosensing
  The 10th Korea-China-Japan Joint Symposium on Enzyme Engineering
  2008年11月04日, 口頭発表（一般）
• Current topics in glycated protein biosensing
  17th indonesian scientific meeting in Japan & Indonesia- Japan friendship forum
  2008年11月01日, 口頭発表（招待・特別）
• Specialized needle-typecholesterol biosensorfor monitoring fish health
  5th World FisheriesCongress
  2008年10月21日, 口頭発表（一般）
• グルコースセンシングルシフェラーゼの開発
  生物工学会
  2008年09月15日, 口頭発表（一般）
• A Novel Enzyme Sensor For Glycated Protein Determination Without Protease Digestion
  Biosensors 2008
  2008年05月12日, 口頭発表（一般）
• 「結合蛋白質を用いた糖化ヘモグロビン測定法の開発」
  メイラード学会
  2007年12月01日, 口頭発表（一般）
• ブリ血漿中糖化タンパク質計測用酵素センサーの開発
  平成19年度日本水産学会秋季大会
  2007年09月26日, 口頭発表（一般）

外部研究資金等

• 魚類の糖化タンパクに関する研究
  共同研究, 自 2004年07月01日, 至 2005年03月31日
• 魚類の糖化タンパク質測定法の研究
  共同研究, 自 2005年07月01日, 至 2006年03月31日
• in vivo評価用酵素電極の開発
  共同研究, 自 2005年09月01日, 至 2006年11月30日
• 魚類の糖化タンパク質測定法の研究
  共同研究, 自 2006年06月20日, 至 2007年03月31日
• 迅速測定用センサに関する研究
  共同研究, 自 2006年09月29日, 至 2007年03月31日
• 新規酵素増幅分析法の開発
  共同研究, 自 2007年04月01日, 至 2008年03月31日
• 魚類の糖化タンパク質に関する研究
  共同研究, 自 2007年05月31日, 至 2008年03月31日
• 臨床診断技術に関する研究
  共同研究, 自 2008年05月22日, 至 2009年05月21日
• 魚類の糖化タンパク質に関する研究
  共同研究, 自 2008年07月14日, 至 2009年03月31日
• 糖化アミノ酸の酵素合成法の開発
  受託研究, 自 2012年04月01日, 至 2012年07月31日
• 糖化アミノ酸の酵素合成法の開発
  受託研究, 自 2011年12月01日, 至 2012年03月31日

委員歴

• 電気化学会
  生物工学研究会事務局
  自 0000, 至 0000
• 日本化学会
  関東支部代議員
  自 201510, 至 201610
• 日本化学会
  関東支部幹事
  自 2013, 至 2015

所属学協会

• 日本メイラード学会
• マリンバイオテクノロジー学会
• 日本薬学会
• 電気化学会
• 農芸化学会
• 生物工学会
• 日本化学会
  自 2015年10月, 至 2016年10月
• 日本化学会
  自 2013年, 至 2015年

受賞

• 電気化学会
  平成２７年度電気化学会女性躍進賞受賞
  新奇酸化還元酵素を応用した電気化学バイオセンシング技術
  2015年03月