Research activity information

• Co-creation and Problem Posing

  Tsuruta Hiroki, Tamaki Hisashi

  現在の大学では、「産学官連携」、「文理融合」、「異分野共創」など、異なるものが相互に関係する状態を表す言葉に触れることが多い。しかし、「連携」、「融合」、「共創」という言葉の意味の違いや「異分野共創」と具体的にどんな関係があるのか等についてはよく分からない。神戸大学では、“知と人を創る「異分野共創研究教育グローバル拠点」を目指す”ことがビジョンの中で掲げられている。異分野共創とは大学や企業などのセクション、さまざまな学問分野など、分野を超えて交流し、新しいモノやコトを創造していくことを指す。異分野共創という言葉をスローガンとして取り扱うだけではなく、その概念を十分に理解し、実際の取り組みに落とし込むことによって新しい価値を生み出していくことが大学に求められていることである。本論文では、問題解決における「問題認識」を題材にして「共創」という概念、そして学問が交わる異分野共創の場である学際領域とは何かを考え、大学の研究や教育に対する影響について論じてみる。

  神戸大学大学教育推進機構, Mar. 2023, 大學教育研究, 31, 131 - 140, Japanese

  Link to Kobe Univ. Repository (Kernel)


• Effects of the expansion of bacterial colonies into the intervillous spaces on the localization of several lymphocyte lineages in the rat ileum

  Hideto Yuasa, Youhei Mantani, Kazuki Miyamoto, Miho Nishida, Masaya Arai, Hiroki Tsuruta, Toshifumi Yokoyama, Nobuhiko Hoshi, Hiroshi Kitagawa

  The effect of bacterial colonies expanded into the intervillous spaces on the localization of several lymphocyte lineages was immunohistochemically investigated in two types of mucosa: ordinary mucosa of rat ileum, which consists of mucosa without any mucosal lymphatic tissue; and follicle-associated mucosa (FAM), which accompanies the parafollicular area under the muscularis mucosae in the rat ileal Peyer's patch. The results showed that bacterial colonies in the intervillous spaces induced increased populations of CD8(+) cells in the epithelium of the intestinal villus in ordinary mucosa (IV) and intestinal villus in FAM (IV-FAM). Bacterial colonies in the intervillous spaces were also associated with increased numbers of IgA(+) cells, which were mainly localized in the lamina propria of basal portions of IV and IV-FAM, and with expanded localization of IgA(+) cells into the villous apex in both IV and IV-FAM. Moreover, IgA(+) cells around the intestinal crypts adjacent to IV or IV-FAM were also increased in response to bacterial colonies. In the IV-FAM, but not IV, L-selectin(+) cells, which were found to be immunopositive for TCR alpha beta or CD19, were drastically increased in the lamina propria from the crypt to middle portion of IV-FAM and in the lumen of central lymph vessel of IV-FAM in response to the bacterial colonies in the intervillous spaces. These findings revealed that the expansion of bacterial colonies into the intervillous spaces accompanies the change of histological localization of the lymphocyte lineage in both the ordinary mucosa and FAM.

  JAPAN SOC VET SCI, Apr. 2019, JOURNAL OF VETERINARY MEDICAL SCIENCE, 81(4) (4), 555 - 566, English, Domestic magazine

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• 神戸大学「志」講義 –新時代を切り拓く羅針盤を身につけるために

  TSURUTA HIROKI, GION KEIKO, OHMURA NAOTO, SAITO MASA-HIKO

  加速度的に変化する社会を生き抜く人材をどう育成していくかが現在の大学が抱える大きな課題である。不確実性の高い現代社会では、解決すべき問題・課題を誰かが与えてくれるわけではなく、学生自らが「問い」を立てて学び、そして解決策を導き、社会の評価を問うというプロセスを試行錯誤しながら進めていかなければならない。このプロセスを回していく原動力が生きるための行動指針である「志」と言える。本論文では、神戸大学が平成 28 年度に開講した「志」講義が必要とされる社会背景、如何にして「志」を醸成させるかの議論および講義の設計、講義の振り返りを紹介する。本講義は、単なる座学形式の講義とは異なり、講師の話を基に学生と講師・コーディネータ教員との対話・議論を展開することで、学生自身が持つ「志」を如何に再確認し、大学生活を含む今後の生活で何とすべきかを考え、共有することを狙いとしている。講義終了後のコミュニケーションシートやアンケートから、その狙い通りに学生自身が自らの「志」をしっかりと考える機会を提供できたと判断した

  神戸大学大学教育推進機構, 2019, 大学教育研究, 27(27) (27), 103 - 112, Japanese

  [Refereed]


• 神戸から配信する遠隔インタラクティブ講義「計算生命科学の基礎」の2017年度報告

  渡邉博文, 鈴木洋介, 八木 学, 石野麻由子, 土井陽子, 江口至洋, TANAKA SHIGENORI, TSURUTA HIROKI, 白井剛, MORI ICHIRO, USUI HIDEYUKI, YOKOKAWA MITSUO

  計算生命科学は，生命の理解に向けて，近年急速に進展している計算科学と医農工理学分野が融合した学際的研究領域である．様々な研究分野や産業界等への研究の拡がりが期待されており，包括的な基礎知識を習得する機会が求められている．神戸大学計算科学教育センターは，関係諸機関と協力して，遠隔インタラクティブ講義「計算生命科学の基礎」シリーズを2014年から全国に配信を開始し，昨年度は600名の受講登録を受け付けた．本稿では，2017年度に実施した「計算生命科学の基礎IV」と，最近注目されているAIやディープラーニングに焦点を当て特別編として実施したディープラーニングチュートリアルの開催結果について報告する．年々受講者が増え続けており，アンケートでも高評価を得ている．．

  Nov. 2018, 大学ICT推進協議会2018年度年次大会論文集, 1 - 4, Japanese

  Symposium


• Ultrastructural and Immunohistochemical Study on the Lamina Propria Cells Beneath Paneth Cells in the Rat Ileum

  Youhei Mantani, Miho Nishida, Kyouji Yamamoto, Kazuki Miyamoto, Hideto Yuasa, Natsumi Masuda, Takuya Omotehara, Hiroki Tsuruta, Toshifumi Yokoyama, Nobuhiko Hoshi, Hiroshi Kitagawa

  Paneth cells secrete bactericidal substances in response to bacterial proliferation on the mucosal surface without directly contacting bacteria. However, the induction mechanism of this transient secretion has not been clarified, although nervous system and/or immunocompetent cells in the lamina propria (LP) might be involved. In this study, we ultrastructurally and immunohistochemically investigated which LP cells are localized beneath Paneth cells and examined the relationship between the Paneth cell-derived cellular processes which extended into the LP and the LP cells. The results showed that various cells-including blood capillary, subepithelial stromal cell, and nerve fiber-were present in the LP beneath Paneth cells. Endothelial cells of blood capillary were the cells most frequently found in this location; they were situated within 1 gm of the Paneth cells and possessed fenestration on the surfaces adjacent to Paneth cells. The Paneth cells rarely extended the cellular processes toward the LP across the basal lamina. Most of the cellular processes of Paneth cells contacted the subepithelial stromal cells. Immunohistochemistry revealed that the CD34(+)CD31(-)alpha SMA(- )stromal cells preferentially localized in the LP beneath the intestinal crypt base, while PDGFR alpha(hi)alpha SMA(+) stromal cells mainly localized around the lateral portions of the intestinal crypt and PDGFR alpha(hi)alpha SMA(-) stromal cells localized in the intestinal villus. From these findings, the existence of blood capillaries beneath Paneth cells might reflect the active exocrine function of Paneth cells. Furthermore, subepithelial stromal cells, probably with a CD34(+)CD31(-)alpha SMA(-)PDGFRa(-/lo) phenotype, beneath the crypt base might affect Paneth cell activity by interacting with their cellular processes. (C) 2018 Wiley Periodicals, Inc.

  WILEY, Jun. 2018, ANATOMICAL RECORD-ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY, 301(6) (6), 1074 - 1085, English, International magazine

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Analysis of major paralogs encoding the Fra a 1 allergen based on their organ-specificity in Fragaria x ananassa

  Misaki Ishibashi, Takeshi Nabe, Yoko Nitta, Hiroki Tsuruta, Miho Iduhara, Yuichi Uno

  Fra a 1 protein in strawberry causes oral allergic syndrome. Over 39 Fra a 1 paralogs have been identified in strawberry genome. Fra a 1.01 is major accumulating protein in edible organs.Strawberry fruits contain allergenic proteins that cause oral allergic syndrome. The hypothesized major allergen is Fra a 1, an ortholog of the birch pollen allergen protein Bet v 1. We organized Fra a 1 genes and analyzed their localizations at the transcriptional and translational levels. In total, 15 new Fra a 1 proteins were identified from the genomic database, increasing the total number of Fra a 1 to 30 proteins encoded by 39 genes. Fra a 1.02 was mostly expressed in receptacles, and Fra a 1.01 in achenes, when analyzed by RNA sequencing. Immunoblotting showed that the Fra a 1.01 protein was broadly accumulated in strawberry organs, while the Fra a 1.02 protein was mostly expressed in receptacles. Recombinant Fra a 1.01 strongly reacted with human IgE. The mRNA and protein expression levels of Fra a 1 did not correlate, indicating the importance of protein levels when evaluating the abundance of allergens in strawberry. Based on the localizations, accumulation levels and reactivity to human IgE, we determined that Fra a 1.01 was the most important allergen, followed by Fra a 1.02, and then other Fra a 1 proteins. The information obtained here will be useful for selecting the target Fra a 1 paralogs when breeding hypoallergenic strawberry.

  SPRINGER, Mar. 2018, PLANT CELL REPORTS, 37(3) (3), 411 - 424, English, International magazine

  [Refereed]

  Scientific journal


• A Report on the Interactive Workshop with “Dialogue Tools for Innovation”

  GION KEIKO, MORI ICHIRO, OHMURA NAOTO, HIRAI MIDORI, TSURUTA HIROKI

  アクティブラーニングやプロジェクトベーストラーニング（PBL）が高等教育に取り入れられて久しい。特に、近年、卓越大学院大学事業、次世代アントレプレナー育成事業（EDGE-NEXT）など、ワークショップ形式を取り入れた講義を提供する機会が増加している。ここでは、「イノベーション対話ツール」を高等教育におけるワークショップ形式の講義の指南書として活用して実施したワークショップの事例を紹介する。本指南書は、産学連携によるイノベーションを目指した対話を推進するために、平成25年度に文部科学省が慶應義塾大学システムデザイン・マネジメント研究科に委託して作成されたものであるが、これを基にワークショップを設計し、ブレインストーミング、親和図法、バリューグラフを実施したところ、学生らに対して課題解決のプロセスにおける発想力並びに対話力の向上と気づきを提供することがで

  Institute for Promotion of Higher Education, Kobe University, Mar. 2018, Kobe Journal of Higher Education, 26(26) (26), 27 - 40, Japanese

  [Refereed]

  Link to Kobe Univ. Repository (Kernel)


• Development of an Education Program for Future Innovators

  TSURUTA HIROKI, GION KEIKO, OHMURA NAOTO

  現代は不確実性の高い社会と呼ばれ、複雑で唯一最適解がない課題が山積されている。さらに科学技術の急激な発展が生活・労働などのヒトが営む活動の環境を加速度的に変化させているのが現代社会の特徴である。このような不確実性の高い社会にこれから出て行く学生は、この社会を変革していく担い手となることが求められている。社会を変革するには、科学技術を基盤とした価値創造、人間を中心にした視点での価値創造による「イノベーション」を引き起こすことが求められる。その担い手が持つ能力は、優れた知識を融合して問題に対峙していける人材である。本論文では、１）現代社会の現状認識とイノベーションを実現する人材の育成の必要性を述べるとともに、２）産業界が求める人材イメージと神戸大学が輩出すべき人材が身につけるべき能力の定義、３）その能力を醸成するための現実社会の問題を題材にした「イノベー

  Institute for Promotion of Higher Education, Kobe University, Mar. 2018, Kobe Journal of Higher Education, 26(26) (26), 119 - 129, Japanese

  [Refereed]


• Interaction between a Unique Minor Protein and a Major Capsid Protein of Bluetongue Virus Controls Virus Infectivity

  Eiko Matsuo, Kiyoshi Yamazaki, Hiroki Tsuruta, Polly Roy

  Among the Reoviridae family of double-stranded RNA viruses, only members of the Orbivirus genus possess a unique structural protein, termed VP6, within their particles. Bluetongue virus (BTV), an important livestock pathogen, is the prototype Orbivirus. BTV VP6 is an ATP-dependent RNA helicase, and it is indispensable for virus replication. In the study described in this report, we investigated how VP6 might be recruited to the virus capsid and whether the BTV structural protein VP3, which forms the internal layer of the virus capsid core, is involved in VP6 recruitment. We first demonstrated that VP6 interacts with VP3 and colocalizes with VP3 during capsid assembly. A series of VP6 mutants was then generated, and in combination with immunoprecipitation and size exclusion chromatographic analyses, we demonstrated that VP6 directly interacts with VP3 via a specific region of the C-terminal portion of VP6. Finally, using our reverse genetics system, mutant VP6 proteins were introduced into the BTV genome and interactions between VP6 and VP3 were shown in a live cell system. We demonstrate that BTV strains possessing a mutant VP6 are replication deficient in wild-type BSR cells and fail to recruit the viral replicase complex into the virus particle core. Taken together, these data suggest that the interaction between VP3 and VP6 could be important in the packaging of the viral genome and early stages of particle formation.IMPORTANCE The orbivirus bluetongue virus (BTV) is the causative agent of bluetongue disease of livestock, often causing significant economic and agricultural impacts in the livestock industry. In the study described in this report, we identified the essential region and residues of the unique orbivirus capsid protein VP6 which are responsible for its interaction with other BTV proteins and its subsequent recruitment into the virus particle. The nature and mechanism of these interactions suggest that VP6 has a key role in packaging of the BTV genome into the virus particle. As such, this is a highly significant finding, as this new understanding of BTV assembly could be exploited to design novel vaccines and antivirals against bluetongue disease.

  AMER SOC MICROBIOLOGY, Feb. 2018, JOURNAL OF VIROLOGY, 92(3) (3), English, International magazine

  [Refereed]

  Scientific journal


• Mechanism of M-cell differentiation accelerated by proliferation of indigenous bacteria in rat Peyer's patches

  Hideto Yuasa, Youhei Mantani, Natsumi Masuda, Miho Nishida, Masaya Arai, Toshifumi Yokoyama, Hiroki Tsuruta, Junichi Kawano, Nobuhiko Hoshi, Hiroshi Kitagawa

  The mechanism by which indigenous bacteria on the follicle-associated epithelium (FAE) of lymphatic follicles (LFs) accelerate the differentiation of microvillous columnar epithelial cells (MV) into M-cells was immunohistochemically investigated in rat Peyer's patches. The results showed that the number of Toll-like receptor (TLR) -4(+) M-cells was greater in the FAE with expansion of bacterial colonies (LFs with bacterial colonies on the FAE: b-LF) than the FAE without expansion of bacterial colonies (nb-LF). TLR-4 was also expressed in the striated borders of MV upstream next to M-cells in the FAE of the b-LF. TLR-4(+) vesicles were frequently detected in the cytoplasms of MV with TLR-4(+) striated borders upstream next to TLR-4(+) M-cells in the FAE of b-LF. These findings suggest that TLR-4(+) MV take up TLR-4 ligands and differentiate into M-cells in the b-LF. Neither the distribution of RANK nor that of RANKL was coincident with that of M-cells in the b-LF. Moreover, RANK, but not RANKL, was expressed in intestinal villi, whereas cleaved caspase-3 was immunonegative in the MV and M-cells of the FAE, unlike in villous epithelial cells. Therefore, RANK/RANKL signaling in the LF might contribute to the down-regulation of epithelial apoptosis to facilitate the differentiation of MV into M-cells in rat Peyer's patches.

  JAPAN SOC VET SCI, Nov. 2017, JOURNAL OF VETERINARY MEDICAL SCIENCE, 79(11) (11), 1826 - 1835, English, Domestic magazine

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• 神戸から配信する遠隔インタラクティブ講義「計算生命科学の基礎」

  渡邉博文, 鈴木洋介, 近藤洋隆, 石野麻由子, 土井陽子, 江口至洋, TANAKA SHIGENORI, TSURUTA HIROKI, 白井剛, MORI ICHIRO, USUI HIDEYUKI, YOKOKAWA MITSUO

  2017, 大学ICT推進協議会2017年度年次大会論文集, TF1-2, 1 - 5, Japanese

  Symposium


• Crystal Structure of Human Herpesvirus 6B Tegument Protein U14

  Bochao Wang, Mitsuhiro Nishimura, Huamin Tang, Akiko Kawabata, Nora F. Mahmoud, Zahra Khanlari, Daizo Hamada, Hiroki Tsuruta, Yasuko Mori

  The tegument protein U14 of human herpesvirus 6B (HHV-6B) constitutes the viral virion structure and is essential for viral growth. To define the characteristics and functions of U14, we determined the crystal structure of the N-terminal domain of HHV-6B U14 (U14-NTD) at 1.85 angstrom resolution. U14-NTD forms an elongated helix-rich fold with a protruding beta hairpin. U14-NTD exists as a dimer exhibiting broad electrostatic interactions and a network of hydrogen bonds. This is first report of the crystal structure and dimerization of HHV-6B U14. The surface of the U14-NTD dimer reveals multiple clusters of negatively- and positively-charged residues that coincide with potential functional sites of U14. Three successive residues, L424, E425 and V426, which relate to viral growth, reside on the beta hairpin close to the dimer's two-fold axis. The hydrophobic side-chains of L424 and V426 that constitute a part of a hydrophobic patch are solvent-exposed, indicating the possibility that the beta hairpin region is a key functional site of HHV-6 U14. Structure-based sequence comparison suggests that U14-NTD corresponds to the core fold conserved among U14 homologs, human herpesvirus 7 U14, and human cytomegalovirus UL25 and UL35, although dimerization appears to be a specific feature of the U14 group.

  PUBLIC LIBRARY SCIENCE, May 2016, PLOS PATHOGENS, 12(5) (5), e1005594, English, International magazine

  Scientific journal


• Different uptake pathways between hydrophilic and hydrophobic compounds in lateral roots of Cucurbita pepo

  Yamazaki Kiyoshi, Tsuruta Hiroki, Inui Hideyuki

  From previous reports, uptake and accumulation of organic compounds by plant roots are strongly related to water solubility. However, the relation between uptake pathways and water solubility remains unclear. Here, we used confocal laser scanning microscopy to observe the uptake of fluorescent hydrophilic and hydrophobic compounds in the living roots of Cucurbita pepo. We found strong regulation of the uptake of berberine, a hydrophobic compound at the endodermal Casparian strip, as previously reported, and similar regulation was also observed in the pericycle. Berberine was loaded into and transported upward through the protoxylem. Perylene, a highly hydrophobic compound, in contrast, passed through the Casparian strip and accumulated preferentially in the endodermis and pericycle. The results of our solvent extraction suggested that perylene diffused into the non-aqueous phase. Therefore, the uptake pathways for the hydrophilic and hydrophobic compoundss are different. These results offer a new way to understand the uptake of agrochemicals and pollutants and to select host plants for phytoremediation.

  Pesticide Science Society of Japan, 2015, Journal of Pesticide Science, 40(3) (3), 99 - 105, English

  [Refereed]


• Expression of Strawberry Allergen Fra a 1 Gene during Fruit Ripening

  D. Futsuki, Y. Nitta, M. Iduhara, H. Tsuruta, T. Tsugehara, Y. Uno

  Consumption of fresh strawberries (Fragaria xananassa) can cause oral allergy syndrome for susceptible individuals. To produce strawberries with low allergen contents a selection of hypoallergenic genotypes and the establishment of culture methods are required. Since several studies have reported that Fra a 1 is a major allergen of strawberry, the expression profiling of the Fra a 1 gene could be an indicator of the allergic potential. In this study, the expression of the Fra a 1 gene was investigated in strawberry during fruit (receptacles and achene) development. The fresh weight and anthocyan content of fruits were also measured to assess their potential correlations with Fra a 1 expression. Fruit of the strawberry were harvested for analyses from a working farm at seven different ripening stages. Real-time PCR revealed the transcript level of the Fra a 1 gene was highest at the early stage of ripening and decreased to approximately 1/70th that level by the red-colored stage. Considering that Fra a 1 belongs to the pathogenesis-related protein 10 family, this result was in line with the report that strawberry fruit may be attacked by oxygen species at the early stage of ripening. Additionally, Fra a 1 gene homologues in Fragaria vesca were surveyed to assess the inducibility of their expression. We found that a gene coding Fra a1 a2 seemed to be expressed in response to stress, and it had major stress-related elements in its promoter region. These results indicate that Fra a 1 expression can be induced by stress, and preharvest treatment to reduce the allergen content will be possible by studying the transcriptional activity of Fra a 1.

  INT SOC HORTICULTURAL SCIENCE, 2014, VII INTERNATIONAL STRAWBERRY SYMPOSIUM, 1049, 323 - 328, English

  [Refereed]

  International conference proceedings


• Varietal Differences in Transcript and Protein Levels of Strawberry Allergen Fra a 1

  Manabu Narukami, Daisuke Futsuki, Takeshi Nabe, Yoko Nitta, Hiroki Tsuruta, Kiyoshi Yamazaki, Miho Iduhara, Yuji Noguchi, Yuichi Uno

  AMER SOC HORTICULTURAL SCIENCE, Sep. 2013, HORTSCIENCE, 48(9) (9), S308 - S309, English

  Research society


• Comparison of IgE Binding Capacity and Expression Analysis of Strawberry Allergen Fra a 1

  Daisuke Futsuki, Takeshi Nabe, Yoko Nitta, Hiroki Tsuruta, Kiyoshi Yamazaki, Miho Iduhara, Yuichi Uno

  AMER SOC HORTICULTURAL SCIENCE, Sep. 2013, HORTSCIENCE, 48(9) (9), S308 - S308, English

  Research society


• IgE reactivity and distribution of strawberry allergen Fra a 1

  FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, NOGUCHI YUJI, UNO YUICHI

  園芸学会, Sep. 2013, 園芸学研究, 12(別冊2) (別冊2), 191, Japanese

  Research society


• Comparison of transcript and protein levels of strawberry allergen Fra a 1 among different cultivars.

  NARUKAMI MANABU, FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, NOGUCHI YUJI, UNO YUICHI

  Veiling Hoogstraten, Sep. 2013, International Strawberry Congress 2013, USB, 頁数6, English

  International conference proceedings


• Analysis of IgE binding capacity and stress inducibility of strawberry allergen Fra a 1.

  FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, UNO YUICHI

  Veiling Hoogstraten, Sep. 2013, International Strawberry Congress 2013, USB, 頁数6, English

  International conference proceedings


• A Major Latex-Like Protein Is a Key Factor in Crop Contamination by Persistent Organic Pollutants

  Hideyuki Inui, Mami Sawada, Junya Goto, Kiyoshi Yamazaki, Noriko Kodama, Hiroki Tsuruta, Heesoo Eun

  This is the first report, to our knowledge, to reveal important factors by which members of the Cucurbitaceae family, such as cucumber (Cucumis sativus), watermelon (Citrullus lanatus), melon (Cucumis melo), pumpkin (Cucurbita pepo), squash (C. pepo), and zucchini (C. pepo), are selectively polluted with highly toxic hydrophobic contaminants, including organochlorine insecticides and dioxins. Xylem sap of C. pepo ssp. pepo, which is a high accumulator of hydrophobic compounds, solubilized the hydrophobic compound pyrene into the aqueous phase via some protein(s). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of xylem sap of two C. pepo subspecies revealed that the amount of 17-kD proteins in C. pepo ssp. pepo was larger than that in C. pepo ssp. ovifera, a low accumulator, suggesting that these proteins may be related to the translocation of hydrophobic compounds. The protein bands at 17 kD contained major latex-like proteins (MLPs), and the corresponding genes MLP-PG1, MLP-GR1, and MLP-GR3 were cloned from the C. pepo cultivars Patty Green and Gold Rush. Expression of the MLP-GR3 gene in C. pepo cultivars was positively correlated with the band intensity of 17-kD proteins and bioconcentration factors toward dioxins and dioxin-like compounds. Recombinant MLP-GR3 bound polychlorinated biphenyls immobilized on magnetic beads, whereas recombinant MLP-PG1 and MLP-GR1 did not. These results indicate that the high expression of MLP-GR3 in C. pepo ssp. pepo plants and the existence of MLP-GR3 in their xylem sap are related to the efficient translocation of hydrophobic contaminants. These findings should be useful for decreasing the contamination of fruit of the Cucurbitaceae family as well as the phytoremediation of hydrophobic contaminants.

  AMER SOC PLANT BIOLOGISTS, Apr. 2013, PLANT PHYSIOLOGY, 161(4) (4), 2128 - 2135, English, International magazine

  Scientific journal


• The temperature dependence of Fra a 1 protein structure.

  NITTA YOKO, FUTSUKI DAISUKE, NABE TAKESHI, TSURUTA HIROKI, IDUHARA MIHO, UNO YUICHI, MATSUO KOICHI

  Jul. 2012, HiSOR Activity Report, 2012, 132 - 133, English

  Research institution


• Complexing of Green Tea Catechins with Food Constituents and Degradation of the Complexes by Lactobacillus plantarum.

  Taeko Hayashi, Shuhei Ueda, Hiroki Tsuruta, Hiroshige Kuwahara, Ro Osawa

  Complexing of green tea catechins with food constituents and their hydrolysis by tannase-producing Lactobacillus plantarum strains, were investigated. Our observations indicated that 1) epigallocatechin gallate (EGCg) and other catechin galloyl esters bound with food ingredients (i.e., proteins) to form a complex that is likely to be unabsorbable through the intestinal wall, whereas most catechins not esterified with gallic acid (GA) remain in free form, not complexing with food ingredients; 2) tannase activity of L. plantarum is strain dependent, possibly grouped into those with high tannase activity hydrolyzing EGCg to epigallocatechin and GA and those with the low activity; and 3) L. plantarum strains with high tannase activity are capable of hydrolyzing not only intact EGCg but also EGCg and other catechin galloyl esters complexed with dietary proteins to free non-galloyl ester catechins and GA. The evidence suggests that L. plantarum with high tannase activity, if it colonizes the human intestine, would release free non-galloyl-ester catechins and GA that are readily absorbed through the human intestinal epithelia from the complexes, thereby ensuring maximum delivery of the bioactive polyphenols of green tea to the host.

  2012, Bioscience of microbiota, food and health, 31(2) (2), 27 - 36, English, Domestic magazine

  [Refereed]

  Scientific journal


• Expression, crystallization and preliminary X-ray diffraction studies of thermostable beta-1,3-xylanase from Thermotoga neapolitana strain DSM 4359

  Fumiyoshi Okazaki, Chiaki Ogino, Akihiko Kondo, Bunzo Mikami, Yoichi Kurebayashi, Hiroki Tsuruta

  Crystals of beta-1,3-xylanase (1,3-beta-D-xylan xylanohydrolase; EC 3.2.1.32) from Thermotoga neapolitana strain DSM 4359 with maximum dimensions of 0.2 x 0.1 x 0.02 mm were grown using the sitting-drop vapour-diffusion method at 293 K over 24 h. The crystals diffracted to a resolution of 1.82 angstrom, allowing structure determination. The crystals belonged to space group P2(1), with unit-cell parameters a = 39.061, b = 75.828, c = 52.140 angstrom; each asymmetric unit cell contained a single molecule.

  INT UNION CRYSTALLOGRAPHY, Jul. 2011, ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 67(Pt 7) (Pt 7), 779 - 781, English, International magazine

  [Refereed]

  Scientific journal


• Polymorphism and distribution of putative cell-surface adhesin-encoding ORFs among human fecal isolates of Bifidobacterium longum subsp longum

  Atsushi Iguchi, Nao Umekawa, Takahiro Maegawa, Hiroki Tsuruta, Toshitaka Odamaki, Jin-Zhong Xiao, Ro Osawa

  The polymorphism of ORFs encoding putative cell-surface adhesins was investigated in Bifidobacterium longum subsp. longum. Firstly, we performed a PCR assay targeting 15 ORFs encoding putative adhesion proteins, which included 8 ORFs with a sortase targeting LPXTG motif, in 42 strains of different pulsotypes isolated from fecal samples from 12 human individuals. We found a variability in the presence of an ORF, BL0675, which encodes a putative fimbrial subunit protein. We sequenced ORFs corresponding to BL0675 in the 42 strains and adjacent ORFs corresponding to BL0674 and BL0676. The results indicated that ORFs corresponding to BL0675 were highly polymorphic with five variant types (i.e. A-, B-, C-, D-, and E-types). Meanwhile, BL0674 and BL0676, which encode an additional putative fimbrial subunit protein and a fimbrial-associated sortase-like protein, were highly conserved. Subsequent quantitative polymerase chain reaction (qPCR) assays targeting the variant types in 89 human fecal samples revealed that A-type was the most commonly distributed (74.2%), followed by B-type (59.6%), D-type (31.5%), E-type (32.6%) and C-type (5.6% prevalence). Since BL0675 is considered to be a fimbrial protein with glycoprotein-binding ability, the proteins encoded by the five variant types of BL0675 may have specific binding properties to various carbohydrate structures expressed on the human intestinal wall, thereby allowing B. longum to colonize the intestine in a host-specific manner.

  SPRINGER, Mar. 2011, ANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY, 99(3) (3), 457 - 471, English, International magazine

  Scientific journal


• Tertiary structure-related activity of tick defensin (persulcatusin) in the taiga tick, Ixodes persulcatus

  Emiko Isogai, Hiroshi Isogai, Kazuhiko Okumura, Hatsuhiro Hori, Hiroki Tsuruta, Yoichi Kurebayashi

  Defensins are small cysteine-rich cationic proteins found in both vertebrates and invertebrates constituting the front line of host innate immunity. To examine the importance of the tertiary structure of tick defensin in its antimicrobial activity, we synthesized two types of the peptides with tertiary structure or primary one on basis of the information of the sequence in the defensin originated from the taiga tick, Ixodes persulcatus. Chemically synthesized peptides were used to investigate the activity spectrum against Staphylococcus aureus, Borrelia garinii and flora-associated bacteria. Both synthetic peptides showed antimicrobial activity against S. aureus in short-time killing within 1 h, but they do not show the activity against B. garinii, Stenotrophomonas maltophila and Bacillus spp., which were frequently isolated from the midgut of I. persulcatus. The teriary structure brought more potent activity to S. aureus than primary one in short-time killing. We also examined its antimicrobial activity by evaluation of growth inhibition in the presence of the synthetic peptides. Minimum inhibitory concentration (MIC) was ranged from 1.2 to 5.0 mu g/ml in tertiary peptide and from 10 to 40 mu g/ml in primary peptide, when 10 strains of S. aureus were used. From the curve of cumulative inhibition rates, MIC50 (MIC which half of the strains showed) to S. aureus is about 1.2 mu g/ml in the peptide with tertiary structure and about 10 mu g/ml in the linear one. Corynebacterium renale is 10 times or more sensitive to tertiary peptide than primary one. In conclusion, the presence of 3 disulfide bridges, which stabilize the molecule and maintain the tertiary structure, is considered to have an effect on their antimicrobial activities against Gram-positive bacteria such as S. aureus.

  SPRINGER, Jan. 2011, EXPERIMENTAL AND APPLIED ACAROLOGY, 53(1) (1), 71 - 77, English, International magazine

  Scientific journal


• Functional Analysis of the Cucumisin Propeptide as a Potent Inhibitor of Its Mature Enzyme

  Masataka Nakagawa, Megumi Ueyama, Hiroki Tsuruta, Tomohide Uno, Kengo Kanamaru, Bunzo Mikami, Hiroshi Yamagata

  Cucumisin is a subtilisin-like serine protease (subtilase) that is found in the juice of melon fruits (Cucumis melo L.). It is synthesized as a preproprotein consisting of a signal peptide, NH2-terminal propeptide, and 67-kDa protease domain. We investigated the role of this propeptide (88 residues) in the cucumisin precursor. Complementary DNAs encoding the propeptides of cucumisin, two other plant subtilases (Arabidopsis ARA12 and rice RSP1), and bacterial subtilisin E were expressed in Escherichia coli independently of their mature enzymes. The cucumisin propeptide strongly inhibited cucumisin in a competitive manner with a K-i value of 6.2 +/- 0.55 nM. Interestingly, cucumisin was also strongly inhibited by ARA12 and RSP1 propeptides but not by the subtilisin E propeptide. In contrast, the propeptides of cucumisin, ARA12, and RSP1 did not inhibit subtilisin. Deletion analysis clearly showed that two hydrophobic regions, Asn(32)-Met(38) and Gly(97)-Leu(103), in the cucumisin propeptide were important for its inhibitory activity. Site-directed mutagenesis also confirmed the role of a Val(36)-centerd hydrophobic cluster within the Asn(32)-Met(38) region in cucumisin inhibition. Circular dichroism spectroscopy revealed that the cucumisin propeptide had a secondary structure without a cognate protease domain and that the thermal unfolding of the propeptide at 90 degrees C was only partial and reversible. A tripeptide, Ile(35)-Val(36)-Tyr(37), in the Asn(32)-Met(38) region was thought to contribute toward the formation of a proper secondary structure necessary for cucumisin inhibition. This is the first report on the function and structural information of the propeptide of a plant serine protease.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Sep. 2010, JOURNAL OF BIOLOGICAL CHEMISTRY, 285(39) (39), 29797 - 29807, English, International magazine

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Crystal Structure of Cold-Active Alkaline Phosphatase from the Psychrophile Shewanella sp.

  Hiroki Tsuruta, Bunzo Mikami, Tetsuo Higashi, Yasuo Aizono

  The crystal structure of a cold-active alkaline phosphatase from a psychrophile, Shewanella sp. (SCAP), was solved at 2.2 angstrom. A refined model showed a homodimer with six metal-ligand sites. The arrangement of the catalytic residues resembled those of alkaline phosphatases (APs), suggesting that the reaction mechanism of SCAP was fundamentally identical to those of other Alps. SCAP had two distinct structural features: (i) a loop with Arg122 that bound to the phosphate moiety of the substrate suffered no constraints from the linkage to other secondary structures, and (ii) Mg3-ligand His109 was considered to undergo repulsive effect with neighboring Trp228. The local flexibility led by these features might be an important factor in the high catalytic efficiency of SCAP at low temperatures.

  TAYLOR & FRANCIS LTD, Jan. 2010, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 74(1) (1), 69 - 74, English, International magazine

  [Refereed]

  Scientific journal


• Identification and cloning of a gene encoding tannase (tannin acylhydrolase) from Lactobacillus plantarum ATCC 14917(T)

  Kazuaki Iwamoto, Hiroki Tsuruta, Yosuke Nishitaini, Ro Osawa

  The gene tanLpl, encoding a novel tannase enzyme (TanLpl), has been cloned from Lactobacillus plantarum ATCC 14917 T. This is the first report of a tannase gene cloned from a bacterial source other than from Staphylococcus lugdunensis, which has been reported elsewhere. The open reading frame of tanLpl, spanning 1410bp, encoded a 469-amino-acid protein that showed 28.8% identity to the tannase of S. lugdunensis with several commonly conserved sequences. These sequences could not be found in putative tannases reported for other bacteria and fungi. TanLpl was expressed in Escherichia coli DH5 alpha from a pGEM-T expression system and purified. SDS-PAGE analysis indicated that purified TanLpl was a monomer polypeptide of approximately 50kDa in size. Subsequent enzymatic characterization revealed that TanLpl was most active in an alkaline pH range at 40 degrees C, which was quite different from that observed for a fungal tannase of Aspergillus oryzae. In addition, the Michaelis-Menten constant of TanLpl was markedly lower than that of A. oryzae tannase. The evidence suggests that TanLpl should be classified into a novel family of tannases. (C) 2008 Elsevier GmbH. All rights reserved.

  ELSEVIER GMBH, URBAN & FISCHER VERLAG, Sep. 2008, SYSTEMATIC AND APPLIED MICROBIOLOGY, 31(4) (4), 269 - 277, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• The role of group bulkiness in the catalytic activity of psychrophile cold-active protein tyrosine phosphatase

  Hiroki Tsuruta, Bunzo Mikami, Chiaki Yamamoto, Hiroshi Yamagata

  The cold-active protein tyrosine phosphatase found in psychrophilic Shewanella species exhibits high catalytic efficiency at low temperatures as well as low thermostability, both of which are characteristics shared by many cold-active enzymes. The structure of cold-active protein tyrosine phosphatase is notable for the presence of three hydrophobic sites (termed the CA, Zn-1 and Zn-2 sites) behind the loop structures comprising the catalytic region. To identify the structural components responsible for specific enzyme characteristics, we determined the structure of wild-type cold-active protein tyrosine phosphatase at high resolution (1.1 angstrom) and measured the catalytic efficiencies of enzymes containing mutations in the three hydrophobic sites. The bulkiness of the amino acid side chains in the core region of the Zn-1 site strongly affects the thermostability and the catalytic efficiency at low temperatures. The mutant enzyme I115M possessed a higher k(cat) at low temperatures. Elucidation of the crystal structure of I115M at a resolution of 1.5 angstrom revealed that the loop structures involved in retaining the nucleophilic group and the acid catalyst are more flexible than in the wild-type enzyme.

  WILEY, Sep. 2008, FEBS JOURNAL, 275(17) (17), 4317 - 4328, English, International magazine

  [Refereed]

  Scientific journal


• Characterisitics and gene cloning of phospholipase D of the psychrophile, Shewanella sp.

  Hiroki Tsuruta, Ryuhei Hayashi, Hiroyuki Ohkawa, Nobuhide Ohkatsu, Masakazu Morimoto, Kaname Nishimoto, Sigeyuki Santou, Yasuo Aizono

  Phospholipase D, with a molecular mass of 64 kDa, was purified from the psychrophile, Shewanella sp. The enzyme showed maximal activity at pH 7.8 and 40 degrees C in the presence of the Ca2+-ion, and its activity at 10 degrees C was 6.5% of maximum. The enzyme exhibited high activity to the non-micelle form of phosphatidylcholine in an aqueous solution containing water miscible alcohols such as methanol, ethanol, iso-propanol, and n-propanol. Nucleotide sequencing of the enzyme gene yielded a deduced amino acid sequence, which showed 36.2% identity to that of Streptomyces chromofuscus phopsholipase D alone. The low sequence similarity to other phopsholipase D enzymes suggests that the purified enzyme might be a novel phospholipase D.

  TAYLOR & FRANCIS LTD, Oct. 2007, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 71(10) (10), 2534 - 2542, English, International magazine

  Scientific journal


• Enzymatic characteristics of cold-active alkaline phosphatase.

  Yamaguchi Haruko, Hiroki Tsuruta, Hiroshi Yamagata, Yasuo Aizono

  Kobe University, 2006, Mem. Grad. School Sci. & Technol., Kobe Univ, 24-A, 23-31, 23 - 31, English

  [Refereed]


• Crystal structure of cold-active protein-tyrosine phosphatase from a psychrophile, Shewanella sp.

  Hiroki Tsuruta, Bunzo Mikami, Yasuo Aizono

  The cold-active protein-tyrosine phosphatase (CAPTPase) of a psychrophile, Shewanella sp., shows high catalytic activity below 20 degrees C. The catalytic residue of CAPTPase is histidine, as opposed to the cysteine of known protein-tyrosine phosphatases (PTPases), and the enzyme protein has three amino acid sequences, Asp-Xaa-His, Gly-Asp-Xaa-Xaa-Asp-Arg and Gly-Asn-His-Glu, that are observed in many protein-serine/threonine phosphatases (PS/TPases). We have determined the crystal structures of CAPTPase at 1.82 angstroms and the enzyme bound with a phosphate ion at 1.90 angstroms resolution using X-ray crystallography and the multiple isomorphous replacement method. The final refined models are comprised of 331 amino acid residues, two metal ions, 447 water molecules, and an acetate or phosphate ion in an asymmetric unit. The enzyme protein consists of three beta-sheets, termed Sheet I, Sheet I', and Sheet II, and 14 alpha-helices. The CAPTPase has a different overall structure from known protein-tyrosine phosphatases. The arrangement of two metal ions, a phosphate ion and the adjacent amino acid residues in the catalytic site of CAPTPase is identical to that of PS/TPases. Thus, it was confirmed that the CAPTPase was a novel PTPase with a conformation similar to the catalytic site of PS/TPase. We speculate that the hydrophobic moiety around the catalytic residue of CAPTPase might play an important role in eliciting high activity at low temperature.

  JAPANESE BIOCHEMICAL SOC, Jan. 2005, Journal of biochemistry, 137(1) (1), 69 - 77, English, International magazine

  [Refereed]

  Scientific journal


• Specification of amino acid residues essential for the catalytic reaction of cold-active protein-tyrosine phosphatase of a psychrophile, Shewanella sp.

  Hiroki Tsuruta, Jun Tamura, Hiroshi Yamagata, Yasuo Aizono

  Protein-tyrosine phosphatase [EC 3.1.3.48] from a psychrophile, Shewanella sp. shows high activity at low temperatures and has the conserved amino acid sequence of protein-Ser/Thr-phosphatases. Site-directed mutagenesis with the conserved amino acid residues indicated that His148 could be important as a general acid catalyst and Asp115 assists the protonation with His148 of the leaving group of a substrate, and that Asp76 and Asp112 were involved in binding to magnesium ions.

  TAYLOR & FRANCIS LTD, Feb. 2004, Bioscience, biotechnology, and biochemistry, 68(2) (2), 440 - 3, English, International magazine

  Scientific journal


• Catalytic efficiency and some structural properties of cold-active protein-tyrosine-phosphatase.

  Hiroki Tsuruta, Yasuo Aizono

  A procedure was established for expression and purification of abundant recombinant cold-active protein-tyrosine-phosphatase (RCPTPase), which showed identical enzymatic characteristics to the native enzyme (NCPTPase). The purified RCPTPase showed high catalytic activity at low temperature and maximal activity at 30 degrees C. RCPTPase has a thermodynamic characteristic in that its activation enthalpy was determined to be low, 4.3 kcal/mol, at temperatures below 19.3 degrees C, where the Arrhenius relationship exhibited an inflection point, in comparison with 20.3 kcal/mol above 19.3 degrees C. Also, the thermostability, DeltaG(water), of the catalytic site in the RCPTPase molecule was increased with a decrease in temperature. It was considered that cold-active protein-tyrosine-phosphatase could maintain its catalytic site in a stable conformation for eliciting high catalytic activity with low activation enthalpy at low temperature.

  Feb. 2003, Journal of biochemistry, 133(2) (2), 225 - 30, English, International magazine

  [Refereed]

  Scientific journal


• Crystallization and preliminary X-ray studies of cold-active protein-tyrosine phosphatase of Shewanella sp.

  Hiroki Tsuruta, Bunzo Mikami, Chiaki Yamamoto, Yasuo Aizono

  Cold-active protein-tyrosine phosphatase (PTPase) of Shewanella sp. was expressed, purified and crystallized using the hanging-drop vapour-diffusion method at two different pH values (4.6 and 8.5). Both crystals are orthorhombic and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 56.4, b = 76.8, c = 81.0 A (pH 8.5) and a = 57.1, b = 77.0 and c = 81.5 A (pH 4.6). Diffraction data to 1.82 A for the pH 8.5 crystal and 2.33 A for the pH 4.6 crystal were collected on a multiwire area detector using a rotating-anode X-ray generator.

  BLACKWELL MUNKSGAARD, Sep. 2002, Acta crystallographica. Section D, Biological crystallography, 58(Pt 9) (Pt 9), 1465 - 6, English, International magazine

  [Refereed]

  Scientific journal


• Cloning of cold-active alkaline phosphatase gene of a psychrophile, Shewanella sp., and expression of the recombinant enzyme

  T Murakawa, H Yamagata, H Tsuruta, Y Aizono

  A psychrophilic alkaline phosphatase (EC 3.1.3.1) from Shewanella sp. is a cold-active enzyme that has high catalytic activity at low temperature [Ishida et al. (1998) Blosci. Blotechnol. Biochem., 62, 2246-2250]. Here, we identified the nucleotide sequence of a gene encoding the enzyme after cloning with the polymerase chain reaction (PCR) and inverted PCR techniques. The deduced amino acid sequence of the enzyme contained conserved amino acids found among mesophilic alkaline phosphatases and showed some structural characteristics including a high content of hydrophobic amino acid residues and the lack of single alpha-helix compared with the alkaline phosphatase of Escherichia coli, which were possibly efficient for catalytic reaction at low temperatures. The recombinant enzyme expressed in E. coli was purified to homogeneity with the molecular mass of 41 kDa. The recombinant enzyme had a specific activity of 1,500 units/mg and had high catalytic activity at low temperatures.

  TAYLOR & FRANCIS LTD, Apr. 2002, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 66(4) (4), 754 - 761, English

  [Refereed]

  Scientific journal


• Cloning of phosphatase I gene from a psychrophile, Shewanella sp., and some properties of the recombinant enzyme

  H Tsuruta, Y Aizono

  Psychrophilic phosphatase I from Shewanella sp. is a cold enzyme that was found as a novel protein-tyrosine-phosphatase (PTPase, EC 3.1.3.48) with a histidine as its catalytic residue [Tsuruta and Aizono (1999) J. Biochem. 125, 690-695]. Here, we determined the nucleotide sequence of a DNA fragment (2,004 bp) containing the phosphatase I gene by cloning with polymerase chain reaction (PCR) and inverted PCR techniques. The deduced amino acid sequence, of the enzyme contained a conserved region of protein-serine/threonine-phosphatase (PPase). The 38.5 kDa-recombinant protein expressed in Escherichia coli was purified to homogeneity by glutathione-Sepharose 4B column chromatography, treatment with endoproteinase and Mono-Q column chromatography. The recombinant enzyme had a specific activity of 49.4 units and, like native psychrophilic phosphatase I, exhibited high catalytic activity at low temperature and PTPase activity.

  JAPANESE BIOCHEMICAL SOC, Jan. 2000, JOURNAL OF BIOCHEMISTRY, 127(1) (1), 143 - 149, English

  [Refereed]

  Scientific journal


• Enzymatical properties of psychrophilic phosphatase I

  H Tsuruta, Y Aizono

  Phosphatase I purified from a psychrophile (Shewanella sp,) [Tsuruta et al. (1998) J. Biochem, 123, 219-225] dephosphorylated O-phospho-L-tyrosine and phospho-tyrosyl residues in phosphorylated poly(Glu(4),Tyr(1)) random polymer (polyEY) and phosphorylated myelin basic protein (MBP) but not phosphoseryl and/or phosphothreonyl residues in phosphorylated histone H1, casein and phosphorylase a, indicating that the enzyme showed protein-tyrosine-phosphatase (PTPase, EC 3.1.3.48)-like activity in vitro. The enzyme was remarkably inhibited by diethylpyrocarbonate (DEPC), monoiodoacetic acid (MIAA), and monoiodoacetamide (MIAM). Binding of 1 mol of DEPC to 1 mol of the enzyme caused complete inhibition of the enzyme; and 0.88 mol of l-carboxymethylated histidine per mole of the enzyme was found when 90% of enzyme activity was lost by modification with C-14-MIAA. These results indicated that this psychrophilic enzyme was a PTPase-like enzyme with histidine as its catalytic residue.

  JAPANESE BIOCHEMICAL SOC, Apr. 1999, JOURNAL OF BIOCHEMISTRY, 125(4) (4), 690 - 695, English

  [Refereed]

  Scientific journal


• Characteristics of psychrophilic alkaline phosphatase

  Y Ishida, H Tsuruta, ST Tsuneta, T Uno, K Watanabe, Y Aizono

  The phosphatase of a psychrophile (Shewanella sp.) was purified by ammonium sulfate fractionation, followed by sequential column chromatographies. The purified enzyme was electrophoretically homogeneous on native- and SDS-PAGE. Its molecular weight was 41,826 by its amino acid composition. The enzyme had its optimal pH for the activity at 9.8, and a broad substrate specificity to dephosphorylate ATP, pyrophosphate, glycerophosphate, and so on. Its activity was increased by metal ions including Mg2+, Mn2+, and Co2+. The maximal activity was observed at 40 degrees C, and the enzyme at 0 degrees C showed 39% of activity at 40 degrees C. The enzyme, however, tended to lose its activity at 20 degrees C and pH 9.8. These results indicated that purified enzyme was an alkaline phosphatase with characteristics; high catalytic efficiency at low temperature and gradual inactivation at an intermediate temperature.

  TAYLOR & FRANCIS LTD, Nov. 1998, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 62(11) (11), 2246 - 2250, English

  [Refereed]

  Scientific journal


• Purification and some characteristics of phosphatase of a psychrophile

  H Tsuruta, ST Tsuneta, Y Ishida, K Watanabe, T Uno, Y Aizono

  The phosphatase of a psychrophile was purified by ammonium sulfate fractionation, and a sequence of chromatographies on DEAE-Cellulofine, butyl-Cellulofine, Sephacryl S-100, and Mono-Q columns, The purified enzyme preparation was found to be electrophoretically homogeneous on native-and SDS-PAGE, and its molecular mass was determined to be 38.4 kDa by MALDI-TOF mass spectrometry, Maximal activity was observed at 30 degrees C and pH 6.0, Furthermore, the activity of this enzyme at 0 and 5 degrees was 27 and 28%, respectively, of that at 30 degrees C, The enzyme was stable in the pH range of 6.0 to 8.0 and up to 20 degrees C. The enzyme was affected by metal ions; the activity was enhanced by Mg2+ and Ca2+ ions, but depressed by Zn2+ ions, Analysis of the amino acid composition indicated that this phosphatase contains no S-S bond, and only a few prolyl residues necessary to retain the rigid structure of a protein molecule, The phosphatase shows typical features of a cold enzyme; high catalytic activity at low temperature and rapid inactivation at an intermediate temperature.

  JAPANESE BIOCHEMICAL SOC, Feb. 1998, JOURNAL OF BIOCHEMISTRY, 123(2) (2), 219 - 225, English

  [Refereed]

  Scientific journal

• 大学インキュベーションの活動と日本版AAASへの期待

  鶴田宏樹

  2021, CBI学会研究講演会, 426th


• 未来の化学工場を考えるプロセスの設計

  祗園景子, 鶴田宏樹, 上田正博, 植田貴志, 神田彰久, 大村直人

  2019, 化学工学会年会研究発表講演要旨集(CD-ROM), 84th


• “メディカル・デバイス・プロデューサー“育成研修プログラムの開発

  保多隆裕, 祇園景子, 鶴田宏樹, 宮崎悟, 水野佳子, 猿渡昌子, 平田健一, 永井洋士, 福本巧

  2019, 日本生体医工学会大会プログラム・抄録集(Web), 58th


• 社会システムと価値変換の役割

  鶴田宏樹, 祗園景子, 大村直人

  2017, 化学工学会秋季大会研究発表講演要旨集(CD-ROM), 49th


• イチゴFra a 1のオーキシン応答性の解析

  石橋美咲, 奈邉健, 新田陽子, 鶴田宏樹, 厳原美穂, 宇野雄一

  2016, 園芸学研究 別冊, 15(1) (1)


• イチゴFra aの花器官における発現解析

  石橋美咲, 奈邉健, 新田陽子, 鶴田宏樹, 厳原美穂, 宇野雄一

  2016, 園芸学研究 別冊, 15(2) (2)


• ジアシルグリセロールキナーゼαの218番目のチロシンリン酸化はドメイン間相互作用と高次構造を変化させる

  脇阪昌明, 岩下智秋, 濱田大三, 濱田大三, 祇園景子, 上田修司, 山之上稔, 鶴田宏樹, 白井康仁

  2015, 日本生化学会大会(Web), 88th


• チロシンリン酸化がジアシルグリセロールキナーゼαのドメイン間相互作用及び高次構造に与える影響

  脇阪昌明, 岩下智秋, 濱田大三, 濱田大三, 祇園景子, 上田修司, 山之上稔, 鶴田宏樹, 白井康仁

  2015, 日本農芸化学会関西支部講演会講演要旨集, 492nd


• 利便性の高い五種混合ワクチンの開発に向けた研究 HHV-6B構造タンパク質の精製および立体構造解析

  鶴田宏樹

  2014, 利便性の高い五種混合ワクチンの開発に向けた研究 平成25年度 総括・分担研究報告書


• C314 Identification of separate pathways for the uptake of hydrophilic and hydrophobic compounds in roots based on in planta observation

  Yamazaki Kiyoshi, Tsuruta Hiroki, Inui Hideyuki

  Pesticide Science Society of Japan, 14 Mar. 2013, 講演要旨集, (38) (38), 149 - 149, Japanese


• イチゴアレルゲンをコードするFra a1遺伝子の単離と発現解析

  夫津木大輔, 奈邉健, 新田陽子, 鶴田宏樹, 厳原美穂, 宇野雄一

  22 Sep. 2012, 園芸学研究 別冊, 11(2) (2), 422, Japanese


• イチゴアレルゲンをコードするFra‐a1遺伝子の果実発達段階における発現解析

  夫津木大輔, 新田陽子, 厳原美穂, 鶴田宏樹, 柘原岳人, 宇野雄一

  28 Mar. 2012, 園芸学研究 別冊, 11(1) (1), 537, Japanese


• C305 Uptake pathways of a hydrophobic compound in plant roots

  Yamazaki Kiyoshi, Tsuruta Hiroki, Inui Hideyuki

  Pesticide Science Society of Japan, 14 Mar. 2012, 講演要旨集, (37) (37), 142 - 142, Japanese


• 海洋性超好熱菌Thermotoga neapolitana由来β-1,3-キシラナーゼの耐熱性を導く要因の解析

  山崎清志, 岡崎文美, 荻野千秋, 近藤昭彦, 三上文三, 榑林陽一, 鶴田宏樹

  2012, 日本農芸化学会大会講演要旨集(Web), 2012


• Functional and structural properties of cold-active enzymes

  鶴田 宏樹, 相薗 泰生

  シーエムシー出版, Jul. 2008, Bio industry, 25(7) (7), 7 - 15,5, Japanese


• Enzymatical properties of enzymes functioning at low temperature

  Hiroki Tsuruta, Yasuo Aizono

  2001, 日本応用酵素協会誌, 36・31-37, Japanese

  Introduction scientific journal


• 好冷菌フォスファターゼの分離精製とその特性 : 酵素

  鶴田 宏樹, 石田 良博, 渡辺 啓一, 相薗 泰生

  社団法人日本農芸化学会, 05 Mar. 1996, 日本農藝化學會誌, 70(0) (0), 305 - 305, Japanese

• 価値創造の教育 : 神戸大学バリュースクールの挑戦

  國部, 克彦, 鶴田, 宏樹, 祗園, 景子

  神戸大学出版会,神戸新聞総合出版センター (発売), Aug. 2021, Japanese, ISBN: 9784909364142


• 価値創造の考え方 : 期待を満足につなぐために

  國部, 克彦, 玉置, 久, 菊池, 誠 (数学)

  日本評論社, Mar. 2021, Japanese, ISBN: 9784535559943

• 未来の化学工場を考えるプロセスの設計

  GION KEIKO, HIROKI TSURUTA, UEDA MASAHIRO, UEDA TAKASHI, KANDA AKIHISA, OHMURA NAOTO

  化学工学会第84回年会, Mar. 2019, Japanese, 芝浦工業大学, Domestic conference

  [Invited]

  Invited oral presentation


• 学生との協創によるアクティブラーニング授業の設計

  GION KEIKO, MATSUI TAKUMI, MIYAGUCHI YUTA, ICHIKAWA KAZUKI, MATSUO TAKUMI, AOKI SARA, KOHAMA NAOHIRO, IGARASHI KAZUNORO, SHIGEKURA YUSUKE, NAKAGAWA SOMA, OHMURA NAOTO, TSURUTA HIROKI

  イノベーション教育学会第6回年次大会, Dec. 2018, Japanese, 東北大学, アクティブラーニングとは，教員からの一方的な講義で知識を覚えるのではなく，学習者が主体的に，仲間と協力しながら課題を解決するような指導・学習方法の総称である．グループワークやディスカッション，体験学習，調査学習など有効とされる． 我々は，アントレプレナー人材育成のプログラムとして，「Creative School」を称する講義を実施ししている．本プログラムの目指す人材像を右図のように定義し，学部学生を対象に「基礎編」と「応用編」を提供している．「基礎編」では，論理的思考，デザイン思考，システム思考をグループワークを通じて習得する授業を学内インフルエンサーである有志学生と共に設計し，全学部1回生を対象にした授業を実施した．有志学生が17回のミーティングを重ね，授業目的，到達目標，ルーブリック評価，授業内容，ロゴマークを作成した．その授業設計のプロセス, Domestic conference

  Poster presentation


• Interaction between a unique minor protein and a major capsid protein of Bluetongue virus controls virus infectivity

  MATSUO EIKO, 山﨑 清志, TSURUTA HIROKI, Polly Roy

  Ninth Interantional Virus Assembly Symposium, May 2018, English, Madeira, Portugal, International conference

  Poster presentation


• Analyzing floral organ-specific expression of Fra a in strawberry.

  ISHIBASHI MISAKI, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, IDUHARA MIHO, UNO YUICHI

  園芸学会平成28年度秋期大会, Sep. 2016, Japanese, 園芸学会, 名城大学（名古屋市）, Domestic conference

  Oral presentation


• イチゴFra a のオーキシン応答性の評価

  ISHIBASHI MISAKI, YOSHIKAWA HIROKI, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, IDUHARA MIHO, UNO YUICHI

  平成28年度園芸学会近畿支部兵庫大会, Aug. 2016, Japanese, 園芸学会近畿支部, 神戸市、神戸大学, Domestic conference

  Oral presentation


• 日本が取り組むべきフューチャー・アースの国際的優先研究テーマの抽出及び研究開発のデザインに関する研究

  谷口 真人, マレー ハイン, 大西 有子, 西村 武司, EBINA KUNIYOSHI, ITOH MASAYUKI, TSURUTA HIROKI, 近藤 康久, 安成 哲三

  Japan Geoscience Union Meeting 2016, May 2016, Japanese, Chiba, Domestic conference

  [Invited]

  Invited oral presentation


• Analyzing organ-specific expression of Fra a in strawberry

  ISHIBASHI MISAKI, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, IDUHARA MIHO, UNO YUICHI

  園芸学会平成27年度秋期大会, Sep. 2015, Japanese, 園芸学会, 徳島大学（徳島市）, Domestic conference

  Oral presentation


• イチゴFra a 1のオーキシン応答性の解析

  ISHIBASHI MISAKI, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, IDUHARA MIHO, UNO YUICHI

  平成27年度園芸学会近畿支部和歌山大会, Aug. 2015, Japanese, 園芸学会近畿支部, 和歌山ビッグ愛, Domestic conference

  Oral presentation


• イチゴアレルゲンFra a 1の発現様式における品種間差異

  NARUKAMI MANABU, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, IDUHARA MIHO, NOGUCHI YUJI, UNO YUICHI

  平成26年度園芸学会近畿支部京都大会, Aug. 2014, Japanese, 園芸学会近畿支部, キャンパスプラザ京都, Domestic conference

  Poster presentation


• IgE reactivity and distribution of strawberry allergen Fra a 1

  FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, NOGUCHI YUJI, UNO YUICHI

  園芸学会平成25年度秋期大会, Sep. 2013, Japanese, 園芸学会, 岩手大学（盛岡市）, Domestic conference

  Oral presentation


• Comparison of transcript and protein levels of strawberry allergen Fra a 1 among different cultivars

  NARUKAMI MANABU, FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, NOGUCHI YUJI, UNO YUICHI

  International Strawberry Congress 2013, Sep. 2013, English, Veiling Hoogstraten, Antwerp (Belgium), International conference

  Poster presentation


• Analysis of IgE binding capacity and stress inducibility of strawberry allergen Fra a 1

  FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, UNO YUICHI

  International Strawberry Congress 2013, Sep. 2013, English, Veiling Hoogstraten, Antwerp (Belgium), International conference

  Oral presentation


• Varietal Differences in Transcript and Protein Levels of Strawberry Allergen Fra a 1

  NARUKAMI MANABU, FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, NOGUCHI YUJI, UNO YUICHI

  American Society for Horticultural Science annual congress 2013, Jul. 2013, English, American Society for Horticultural Science, Palm Desert (California, USA), International conference

  Poster presentation


• Comparison of IgE Binding Capacity and Expression Analysis of Strawberry Allergen Fra a 1

  FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, UNO YUICHI

  American Society for Horticultural Science annual congress 2013, Jul. 2013, English, American Society for Horticultural Science, Palm Desert (California, USA), International conference

  Poster presentation


• イチゴアレルゲンをコードするFra a 1 遺伝子の解析

  FUTSUKI DAISUKE, NABE TAKESHI, NITTA YOKO, TSURUTA HIROKI, YAMAZAKI KIYOSHI, IDUHARA MIHO, UNO YUICHI

  神戸大学研究基盤センター 若手フロンティア研究会２０１２, Dec. 2012, Japanese, 神戸大学研究基盤センター, 神戸大学百年記念館, Domestic conference

  Poster presentation


• ブルータングウイルス（BTV）コアタンパク質VP3とVP6の結合に関する研究

  MATSUO EIKO, 山崎 清志, Esther Leon, SAEKI KEIICHI, KAWANO JUNICHI, Steve Matthew, TSURUTA HIROKI, Polly Roy

  第65回日本細菌学会関西支部総会, Nov. 2012, Japanese, 神戸, Domestic conference

  Oral presentation


• ブルータングウイルス（BTV）コアタンパク質VP3とVP6の結合に関する研究

  MATSUO EIKO, Esther Leon, 山崎 清志, SAEKI KEIICHI, KAWANO JUNICHI, Steve Matthew, TSURUTA HIROKI, Polly Roy

  第60回日本ウイルス学会学術集会, Nov. 2012, Japanese, 大阪, Domestic conference

  Oral presentation


• 海洋性超好熱菌Thermotoga neapolitana由来β-1,3-キシラナーゼの耐熱性を導く要因の解析

  山崎 清志, 岡崎 文美, 荻野 千秋, 近藤 昭彦, 三上 文三, 榑林 陽一, 鶴田 宏樹

  日本農芸化学会2012年度大会, Mar. 2012, Japanese, 日本農芸化学会, 京都市, Domestic conference

  Oral presentation


• 低温活性protein tyrosine phosphataseの機能発現に寄与する構造要因

  Hiroki Tsuruta, Bunzo Mikami

  日本農芸化学会2006年度大会, 2006, Japanese, 日本農芸化学会, 京都, Domestic conference

  Oral presentation


• 低温活性protein-tyrosine- phosphataseの機能発現機構

  鶴田 宏樹, 山本 千晶, 三上 文三, 山形 裕士, 相薗 泰生

  日本農芸化学会大会2005年度大会 要旨集p.42, Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

  Oral presentation


• 低温活性protein-tyrosine-phosphataseの機能発現機構

  Hiroki Tsuruta, Chiaki Yamamoto, Bunzo Mikami, Hiroshi Yamagata, Yasuo Aizono

  日本農芸化学会2005年度大会, 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

  Poster presentation


• 低温活性protein-tyrosine-phosphataseの機能特性及び構造特性

  Hiroki Tsuruta, Bunzo Mikami, Yasuo Aizono

  日本分子生物学会2005年度年会, 2005, Japanese, 日本分子生物学会, 福岡, Domestic conference

  Invited oral presentation


• フラボノール小腸吸収時における新規の代謝経路について

  Atsumi Mori, Shoko Habuchi, Hiroki Tsuruta, Takashi Hashimoto, Kazuki Kanazawa

  日本農芸化学会, 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

  Poster presentation


• 低温活性protein-tyrosine phosphatase (PTPase)の触媒部位近傍に位置する疎水性部位の活性発現に対する寄与

  Chiaki Yamamoto, Hiroki Tsuruta, Hiroshi Yamagata, Yasuo Aizono

  日本農芸化学会2004年度大会, Mar. 2004, Japanese, 日本農芸化学会, 広島, Domestic conference

  Oral presentation


• 低温活性Phospholipase Dの大量発現系の確立と機能解析

  Hiroyuki Ohkawa, Hiroki Tsuruta, Hiroshi Yamagata, Yasuo Aizono

  日本農芸化学会2004年度大会, Mar. 2004, Japanese, 日本農芸化学会, 広島, Domestic conference

  Oral presentation


• 低温活性protein-tyrosine phosphatae (PTPase) の触媒部位近傍に位置する疎水性部位の活性発現に対する寄与

  山本 千晶, 鶴田 宏樹, 山形 裕士, 相薗 泰生

  日本農芸化学会2004年度大会, 2004, Japanese, 日本農芸化学会, 未記入, Domestic conference

  Oral presentation


• 低温活性Phospholipase D の大量発現系の確率と機能解析

  大川 寛幸, 鶴田 宏樹, 山形 裕士, 相薗 泰生

  日本農芸化学会2004年度大会, 2004, Japanese, 日本農芸化学会, 未記入, Domestic conference

  Oral presentation


• フラボノール類新規代謝産物アミノ体に関する研究

  羽渕 祥子, 森 敦美, 鶴田 宏樹, 芦田 均, 金沢 和樹

  2004年度日本農芸化学会本大会, 2004, Japanese, 日本農芸化学会, 広島, Domestic conference

  Oral presentation


• フラボノール類の新規代謝産物アミノ体に関する研究

  Shoko Habuchi, Atsumi Mori, Hiroki Tsuruta, Hitoshi Ashida, Kazuki Kanazawa

  日本農芸化学会2004年度大会, 2004, Japanese, 日本農芸化学会, 広島, Domestic conference

  Oral presentation


• Hydrophobic interaction responsible for constructing the active center of cold-active protein-tyrosine phosphatase.

  山本 千晶, 鶴田 宏樹, 山形 裕士, 相薗 泰生

  日本生化学会第76回大会、講演要旨集, Oct. 2003, English, 日本生化学会, パシフィコ横浜, Domestic conference

  Oral presentation


• 低温活性プロテイン-チロシン-フォスファターゼのX線結晶構造解析

  Hiroki Tsuruta, Bunzo Mikami, Yasuo Aizono

  日本農芸化学会2003年度大会, 2003, Japanese, 日本農芸化学会, 東京, Domestic conference

  Oral presentation


• Hydrophobic interaction responsible for constructing the active center of cold-active protein-tyrosine phosphatase

  Chiaki Yamamoto, Hiroki Tsuruta, Hiroshi Yamagata, Yasuo Aizono

  日本生化学会2003年度年会, 2003, Japanese, 日本生化学会, 横浜, Domestic conference

  Poster presentation


• 低温活性アルカリフォスファターゼの大腸菌における発現及び発現酵素の特性

  Tetsuo Higashi, Hiroki Tsuruta, Yasuo Aizono

  日本農芸化学会2002年度大会, 2002, Japanese, 日本農芸化学会, 仙台, Domestic conference

  Invited oral presentation


• Psychrophilic Phospholipase Dの分子特性

  Ryuhei Hayashi, Hiroki Tsuruta, Hiroyuki Ohkawa, Yasuo Aizono

  日本農芸化学会2002年度大会, 2002, Japanese, 日本農芸化学会, 仙台, Domestic conference

  Oral presentation


• 低温プロテイン-チロシン-フォスファターゼの分子特性

  Hiroki Tsuruta, Yasuo Aizono

  日本農芸化学会2001年度大会, 2001, Japanese, 日本農芸化学会, 京都, Domestic conference

  Oral presentation


• 低温アルカリフォスファターゼの分子機能

  Haruko Yamaguchi, Hiroki Tsuruta, Yasuo Aizono

  日本農芸化学会2001年度大会, 2001, Japanese, 日本農芸化学会, 京都, Domestic conference

  Oral presentation


• 低温アルカリフォスファターゼの遺伝子クローニング及び組換えタンパク質の大腸菌における発現とその特性解析

  Takeshi Murakawa, Hiroki Tsuruta, Hiroshi Yamagata, Yasuo Aizono

  日本農芸化学会2001年度大会, 2001, Japanese, 日本農芸化学会, 京都, Domestic conference

  Oral presentation


• 低温ホスホリパーゼDの分離精製と機能

  Nobuhide Ohkatsu, Hiroki Tsuruta, Yasuo Aizono

  日本農芸化学会200年度大会, 2000, Japanese, 日本農芸化学会, 東京, Domestic conference

  Poster presentation


• Psychrophilic Phosphatase I遺伝子のクローニングとその組換え酵素の特性

  Hiroki Tsuruta, Yasuo Aizono

  日本農芸化学会2000年度大会, 2000, Japanese, 日本農芸化学会, 東京, Domestic conference

  Poster presentation


• Psychrophilic Phosphatase Iの酵素的性質

  Hiroki Tsuruta, Yasuo Aizono

  日本農芸化学会1999年度大会, 1999, Japanese, 日本農芸化学会, 福岡, Domestic conference

  Poster presentation


• 好冷菌フォスファターゼIIの分離精製とその性質

  Yoshihiro Ishida, Hiroki Tsuruta, Keiichi Watanabe, Yasuo Aizono

  日本農芸化学会1997年度大会, 1997, Japanese, 日本農芸化学会, 東京, Domestic conference

  Oral presentation


• Phosphatase I（低温酵素）の分子特性

  Hiroki Tsuruta, Tomohide Uno, Keiichi Watanabe, Yasuo Aizono

  日本農芸化学会西日本・関西合同大会, 1997, Japanese, 日本農芸化学会西日本・関西支部, 佐賀, Domestic conference

  Oral presentation


• 好冷菌フォスファターゼの分離精製とその特性

  Hiroki Tsuruta, Yoshihiro Ishida, Keiichi Watanabe, Yasuo Aizono

  日本農芸化学会1996年度大会, 1996, Japanese, 日本農芸化学会, 京都, Domestic conference

  Oral presentation

• 極限微生物学会


• 日本分子生物学会


• 日本蛋白質科学会


• 日本生化学会


• 日本農芸化学会

• 「道場」の機能に基づく持続可能な科学コミュニケーションの場づくり

  祇園 景子, 大村 直人, 鶴田 宏樹

  日本学術振興会, 科学研究費助成事業, 基盤研究(C), 神戸大学, 01 Apr. 2020 - 31 Mar. 2023

  近代の道場は，正式な寺院や僧侶が近くにないときに，日常的な法事や仏教行事を担う人あるいは場所となった．出家信者と在家信者の間に位置する檀家総代であるとともに，在家信者たちが集まる場所をさす．道場の一つの役割である仏教行事の挙行に着目し，行事が一般市民の科学的興味・関心に与える影響を検討した．具体的には，祭りを通じて構築される人とのつながりと防災意識との関係を調査した．祭りの運営に携わる人，祭りに行く人，行かない人について，消防団員とそうでない人たちに分けて，各々の防災意識の違いをソーシャル・キャピタルの観点から分析した．ソーシャル・キャピタルとは，社会・地域における人々の信頼関係やつながりを表す概念で，共通の目的に向けて協調行動を促し，社会の効率性や成長，持続性を高める社会的資源を意味する．アンケート調査を実施したところ，消防団員であろうとなかろうと，祭りの運営に携わる人のほうが祭りへ行くだけの人よりも，ハザードマップや避難場所・経路を理解していたり，備蓄品を準備したり，近所の要支援者を把握していたり，防災意識が高い傾向にあることが分かった．また，消防団員ではない人で祭りに行く人は，ソーシャル・キャピタルがハザードマップや避難場所・経路を理解したり，備蓄品を準備するなど，防災における自助意識に与える影響が大きいことが判明した．消防団員ではない人たちを非専門家とすると，ソーシャル・キャピタルを高めることによって，非専門家の科学的興味・関心を向上させることができることを示唆している．


• Building an ecosystem for human development in rural area

  中塚 雅也

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (B), Kobe University, Apr. 2018 - Mar. 2021

  A人材育成エコシステムのモデル確立については，兵庫県丹波篠山市におけるアクションリサーチを展開した。特に，これまでの5期に渡るイノベーターズスクールの活動の評価分析を行い，課題を抽出した。また，これを神戸市にモデル移転し「神戸農村スタートアッププログラム」として実践した。一方，兵庫県たつの市における空き屋活用の事例から，NPOと市民出資の不動産会社を核とする多様なアクターネットワークの実態と要点を明らかにした。なお，兵庫県下の新規就農者を対象とした分析も行実施し，その定着に地域差，つまりエコシステムの差がある可能性を示唆した。 B地域づくりリーダーの特性と地域力醸成要件の解明については，大分県において実践的な調査をすすめた。宮崎県大崎市との比較調査しながら，地域運営組織に対する市役所および中間支援組織による支援課題を明らかにした。また，佐渡島におけるボトムアップ型のコミュニティ開発の事例分析を行い，その支援体制を整理した。一方，畦畔管理作業やため池管理については，非農家や非住民の参画を可能とした組織づくりの要点と課題を明らかにした。さらに，小学校から高校までの総合的学習（探求）による学校教育，公民館活動による社会教育，関係人口づくりなど多面的な対応が実質化していることを確認し，その際には，多様な主体がミックスされる場と仕組みが重要であることを明らかにした。 C農村における起業家の創出・成長要件の解明については，丹波篠山市を対象に地理的条件に焦点をあてた分析を進めた。また，多くの起業人材を輩出している島根県邑南町と島根県西粟倉村を事例として取り上げ，移住と起業を促す地域システムの実態を把握した。継業に関しては，鳥取市吉岡温泉町を事例に，鳥取県，地域おこし協力隊，NPO法人学生人材バンクと協働して，基礎的な実態調査およびセミナーを行い，継業を取り巻く課題整理を行った。

  Competitive research funding


• Development of super-resolution structure analysis techniques for bio-macromolecules based on a quantum chemical calculation

  Fukuzawa Kaori, HONMA Teruki, TANAKA Shigenori, TSURUTA Hiroki, WATANABE Chiduru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), 01 Apr. 2015 - 31 Mar. 2018

  In this study, we are developing a new precise structure determination method of protein - ligand complex by using X-ray crystal structure analysis and quantum chemical calculation. Based on the fragment molecular orbital method, we attempted to use the energy calculation, structure optimization, interaction analysis, and electron density analysis to substantially increase the resolution of structural analysis by experiment. t was shown that quantum chemical calculations are quite useful in predicting high-precision activity of Pim 1 kinase inhibitors, hydration of anti-influenza drugs, prediction of high-resolution structures from low-resolution crystal structures of estrogen receptors.


• Comprehensive investigation of molecular mechansim of intracellular localization of DG kinase as an example of general enzmes

  Shirai Yasuhito, TSURUTA HIROKI

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Kobe University, 01 Apr. 2015 - 31 Mar. 2018

  In this study, molecular mechanism of nuclear-cytoplasm shuttling of diacylglycerol kinase alpha (DGKa) was investigated. We found that RVH-EF region directly bound to C1 domain which functions as nuclear localization signal (NLS) nteraction was enhanced by tyrosin phosphorylation at Y218, resulting weaken the function of C1 domain as NLS. We also identified cyclin dependent kinase 12 and Rheb as binding proteins of DGKa in the nucleus and the cytoplasm, respectively. The former seems to work to stop the cell cycle and the latter seems to contribute the proliferation.


• Development of theoretical methods for interaction analysis of protein-ligand systems in computational drug design

  TANAKA Shigenori, TSURUTA Hiroki

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Kobe University, 01 Apr. 2014 - 31 Mar. 2017

  In order to construct a platform of the structure-based insilico drug design, ab initio large-scale quamtum-chemical calculations for protein-ligand systems were carried out on the basis of the fragment molecular orbital (FMO) method. As one of our activities in the "FMO Drug Design Consortium" for the collaborations of industry, academia and government, the interaction analyses for cancer-related kinase p38 in complex with about one-hundred species of ligand molecules were performed, and the data base of the calculated results was constructed. In addition, the molecular dynamics simulations for the state transitions of cancer-related Ras protein were also carried out to investigate the open-close mechanism of molecular switches, which has provided useful information for the design of effective inhibitors for Ras.


• Development of an evaluation system for inducibility of strawberry allergy.

  UNO Yuichi, NABE Takeshi, NITTA Yoko, TSURUTA Hiroki

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Challenging Exploratory Research, Kobe University, Apr. 2012 - Mar. 2014

  Fruit allergies are serious health problems that limit the quality of life and slumping consumption of agricultural products. Strawberry can cause oral allergy syndrome, an allergic reaction in patients. In this study, the fundamental knowledge was obtained by analyzing strawberry allergen Fra a and antiallergic constituents, towards the development of an evaluation system for inducibility of strawberry allergy. The analysis of IgE binding capacity revealed that Fra a 1 might plays an important role in determining the allergenicity of strawberry fruits. Fra a 1 content varied by cultivars, cultivating methods, ripening stages, and different tissues. The secondary structure of Fra a 1 was altered by heating to more than 60 C. There is a difference in antiallergic effects among strawberry cultivars. These findings suggest that strawberry allergy symptoms could be reduced by appropriate selection of cultivars, cultivating methods, and cooking methods.

  Competitive research funding


• Development of monitoring and remediation of persistent organic pollutants using constituents of xylem sap and root exudates from Cucurbitaceae family

  INUI Hideyuki, EUN Heesoo, TSURUTA Hiroki

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (A), Kobe University, 2011 - 31 Mar. 2014

  We tried to clarify uptake mechanisms of persistent organic pollutants, such as dioxins, by Cucurbitaceae family. Transporting proteins in xylem sap and organic acids in root exudates were responsible for its uptake to plants. Expression of genes related to uptake in plants led to dioxin accumulation in the aerial parts of plants. These results showed that phytoremediation systems for persistent pollutants are developed by the use of functions derived from Cucurbitaceae family.

  Competitive research funding


• A-STEP「麹菌由来ペプチドをリード化合物とした構造科学に基づく新規薬剤候補物質の開発」

  鶴田 宏樹

  研究成果最適展開支援プログラム フィージビリティスタディステージ 探索タイプ 拠点型, 神戸大学, 2011 - 2011, Principal investigator

  麹菌Aspergillus oryzaeはエラスターゼ活性を阻害するペプチド(A.oryzae Elastase Inhibitor、AOEI)を産生する。AOEIは新規阻害剤として将来的に産業利用できる可能性を有するものの、その阻害様式などは不明である。そこで生化学的・構造生物学的アプローチによって、AOEIの阻害特性を解明することを計画した。AOEIの生体内発現量は少ないため、組換えAOEIの取得が研究の律速となっている。本研究では、特性解析に必要な量のAOEIを取得するための、大腸菌を用いた発現系を確立した。今後、この組換えAOEIを用いた構造ベースでの新規阻害剤開発を行う予定である。

  Competitive research funding


• Environmental factor for cold-adaptation of psychrophilic metalloenzyme

  TSURUTA Hiroki

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Young Scientists (B), Kobe University, 2008 - 2010, Principal investigator

  For the construction of structure leading high cold activity of psychrophilic phosphatase, the magnesium concentration during folding of this enzyme protein was an important environmental factor.

  Competitive research funding


• Research of the structural factors for eliciting the highly catalytic efficiency of cold-active enzyme

  TSURUTA Hiroki, MIKAMI Bunzo

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Kobe University, 2005 - 2006, Principal investigator

  Our final goal is that the investigation of the structural factor(s) for eliciting the highly catalytic efficiency of cold-active enzymes. For the purpose, we focused on cold-active protein-tyorosine phopshtase (CAPTPase) of a psychrophile Shewanella sp. as a experimental material. CAPTPase is a novel protein tyrosine phosphatase with the phosphoesterase motif conserved among metallophosphoesterases including many protein serine/threonine phosphatases. In this study, firstly, in order to reveal the detailed structural features and the reaction mechanism of this enzyme, we specified the metal ions in the enzyme protein and determined the crystal structure of CAPTPase at resolution of 1.1 Å. As the results, it was obvious that CAPTPase had two Zn atoms (termed Zn1 and Zn2) constructing a dinuclear Zn-Zn center in the catalytic site and that the hydrolysis of this enzyme was initiated by the nucleophilic attack with a water molecule bridging two Zn atoms after the substrate binding. The loop structures in the dinuclear Zn-Zn center were lined mainly with hydrophobic interactions. The mutational analysis for these hydrophobic residues and the determined crystal structure of I115M mutant enzyme, of which the catalytic efficiency was higher than that of the wild type enzyme, indicated that the catalysis of CAPTPase was dependent on the chemical bond force of hydrophobic interactions behind the loop structures contributing to the configuration of Zn1 atom.

  Competitive research funding


• 低温酵素(フォスファターゼ)の分子機能に関する研究

  鶴田 宏樹

  日本学術振興会, 科学研究費助成事業, 特別研究員奨励費, 神戸大学, 2000 - 2001