2002, 北海道大学クラーク記念財団, 海外派遣助成　　　　　　　　　　　　　　　
鷲尾 健司

Gene cloning and characterization of a vanadium-dependent bromoperoxidase from the red alga Laurencia saitoi, a producer of brominated diterpenoids and triterpenoids
Kensuke Kaneko, Daiki Kobayashi, Shiro Masaki, Kenji Washio, Masaaki Morikawa, Tatsufumi Okino
JOURNAL OF APPLIED PHYCOLOGY, Apr. 2023
English, Scientific journal

Nostosin G and Spiroidesin B from the Cyanobacterium Dolichospermum sp. NIES-1697
Chin-Soon Phan, Jakia Jerin Mehjabin, Andrea Roxanne J. Anas, Masahiro Hayasaka, Reiko Onoki, Juting Wang, Taiki Umezawa, Kenji Washio, Masaaki Morikawa, Tatsufumi Okino
Journal of Natural Products, American Chemical Society (ACS), 10 Aug. 2022
Scientific journal

Characterization of vanadium-dependent bromoperoxidases involved in the production of brominated sesquiterpenes by the red alga Laurencia okamurae
Takafumi Ishikawa, Kenji Washio, Kensuke Kaneko, Xiao Rong Tang, Masaaki Morikawa, Tatsufumi Okino
Applied Phycology, 3, 1, 120, 131, Informa UK Limited, 20 Jul. 2022, [Peer-reviewed]
English, Scientific journal

Wewakazole B, a Cytotoxic Cyanobactin from the Cyanobacterium Moorea producens Collected in the Red Sea
Julius Adam V. Lopez, Sultan S. Al-Lihaibi, Walied M. Alarif, Ahmed Abdel-Lateff, Yasuyuki Nogata, Kenji Washio, Masaaki Morikawa, Tatsufumi Okino
JOURNAL OF NATURAL PRODUCTS, 79, 4, 1213, 1218, Apr. 2016, [Peer-reviewed]
English, Scientific journal

An N-terminal region of a Myb-like protein is involved in its intracellular localization and activation of a gibberellin-inducible proteinase gene in germinated rice seeds
Keita Sutoh, Kenji Washio, Ryozo Imai, Masamitsu Wada, Tomonori Nakai, Daisuke Yamauchi
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 79, 5, 747, 759, May 2015, [Peer-reviewed]
English, Scientific journal

Wewakazole B, A New Cyclic Dodecapeptide from the Red Sea Cyanobacterium Moorea producens
Lopez Julius Adam Velasco, Al-Lihaibi Sultan S., Alarif Walied M., Abdel-Lateff Ahmed, Washio Kenji, Morikawa Masaaki, Okino Tatsufumi
Symposium on the Chemistry of Natural Products, symposium papers, 57, PosterP23, Symposium on the Chemistry of Natural Products Steering Committee, 2015
Japanese, Cyanobacteria are prolific sources of novel bioactive compounds. The chemical profiles we obtained using LC-MS and checked with the MarinLit database showed that there are potential new compounds from Moorea producens samples obtained from the Red Sea. In this presentation the isolation and structure elucidation of a new cyclic dodecapeptide, wewakazole B (1), is reported.

The brownish-red filamentous sample was obtained by SCUBA near Jeddah, Saudi Arabia. A small portion of the sample was preserved in RNAlater and identified as M. producens by standard 16S rRNA gene sequencing using cyanobacteria-specific primers 106F and 1509R, and internal primers 359F and 781R (Martinez-Murcia et al., 1995; Nubel et al., 1997). The bulk of the sample was extracted with methanol followed by solvent partition using EtOAc and water, then, BuOH. These extracts were tested for antifouling activity against the barnacle larvae Amphibalanus amphitrite, cytotoxicity against MCF-7 breast cancer cells, and trypsin inhibition. LC-MS was carried out in parallel for dereplication in conjunction with the MarinLit database. The extracts were found to be inactive but a unique [M+H]⁺ m/z of 1127 was detected in the EtOAc extract and was pursued. A series of open column chromatography and HPLCs afforded 0.4 mg of 1.

The molecular formula, C₅₈H₇₀N₁₂O₁₂, determined by HRESIMS ([M+H]⁺ m/z1127.5310, ∆ = 0.39 ppm), suggested thirty unsaturations. Dereplication using mass and ¹H NMR data showed wewakazole (Nogle et al., 2003) as the nearest candidate. HSQC, COSY, and TOCSY revealed nine common and three modified amino acid residues which were connected by HMBC and ROESY correlations acquiring five fragments. Connections to the oxazole and methyloxazole fragments proved to be a challenge due to the lack of correlations towards neighboring amino acids. All possible combinations were considered that lead to the most logical planar structure and cyclic sequence. Chiral LC-MS and HPLC analysis showed that all amino acid residues possess the L- configuration. The wewakazoles are the only cyclic peptides from marine cyanobacteria having both Oxz and MeOxz. Their function and bioactivity require further investigation.

cDNA cloning and characterization of vanadium-dependent bromoperoxidases from the red alga Laurencia nipponica
Kensuke Kaneko, Kenji Washio, Taiki Umezawa, Fuyuhiko Matsuda, Masaaki Morikawa, Tatsufumi Okino
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 78, 8, 1310, 1319, Aug. 2014, [Peer-reviewed]
English, Scientific journal

Bromoperoxidases from Laurencia species
Kaneko Kensuke, Washio Kenji, Umezawa Taiki, Kobayashi Daiki, Tang Xiaorong, Matsuda Fuyuhiko, Morikawa Masaaki, Okino Tatsufumi
Symposium on the Chemistry of Natural Products, symposium papers, 56, Poster53, Symposium on the Chemistry of Natural Products Steering Committee, 2014
Japanese, The genus of red alga Laurencia is one of the richest producers of halogenated compounds (mostly brominated) ³⁾. Murai et al ⁴⁾ and Butler et al ⁹⁾ showed enzymatic bromination of Laurencia C₁₅acetogenins including laurencin (1) ^1,2)and sesquiterpenoids snyderols using partially purified bromoperoxidase (BPO) from L. nipponica and vanadium-dependent BPO (VBPO) from a mixture of red algae including L. pacifica. However, since the lack of information of sequences, the structural information and biochemical aspects of BPO from L. nipponica and VBPO from L. pacifica remains unknown. Here, we report cDNA cloning of VBPOs from L. nipponicaand in vitro enzymatic bromination of putative biosynthetic precursors using recombinant enzymes.

Two VBPOs (LnVBPO1 and LnVBPO2) were cloned from L. nipponica and recombinant LnVBPOs were prepared. Recombinant LnVBPOs were subjected to enzyme activity assay. A general substrate, monochlorodimedone (MCD), was utilized for K_m determination for H₂O₂ and , essential for bromination of organic substrate. K_m for H₂O₂was 0.03 mM in both LnVBPOs, K_m for were 0.53 mM for LnVBPO1 and 0.35 mM for LnVBPO2, respectively. A derivative of putative precursor of laurencin (1), TMS-capped (3E, 6R, 7R)-laurediol (4) was subjected to LnVBPOs enzymatic bromination and gave similar MS/MS spectrum of deacetylaurencin (3). Since the compound 3 is considered to be the final precursor of 1, it is suggested that VBPO catalyzes bromination in Laurenciametabolites.

We also cloned VBPOs from other Laurencia spp. which produce brominated sesquiterpenoids (L. okamurae) and triterpenoids (L. saitoi). We will discuss in vitro enzymatic bromination of these two LaurenciaVBPOs.

Efficacy of forming biofilms by naphthalene degrading Pseudomonas stutzeri T102 toward bioremediation technology and its molecular mechanisms
Kohei Shimada, Yoshikane Itoh, Kenji Washio, Masaaki Morikawa
CHEMOSPHERE, 87, 3, 226, 233, Apr. 2012, [Peer-reviewed]
English, Scientific journal

A Truncated Form of SpoT, Including the ACT Domain, Inhibits the Production of Cyclic Lipopeptide Arthrofactin, and Is Associated with Moderate Elevation of Guanosine 3 ',5 '-Bispyrophosphate Level in Pseudomonas sp MIS38
Kenji Washio, Siew Ping Lim, Niran Roongsawang, Masaaki Morikawa
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 75, 10, 1880, 1888, Oct. 2011, [Peer-reviewed]
English, Scientific journal

Sustainable Biodegradation of Phenol by Acinetobacter calcoaceticus P23 Isolated from the Rhizosphere of Duckweed Lemna aoukikusa
Fumiko Yamaga, Kenji Washio, Masaaki Morikawa
ENVIRONMENTAL SCIENCE & TECHNOLOGY, 44, 16, 6470, 6474, Aug. 2010, [Peer-reviewed]
English, Scientific journal

Identification and Characterization of the Genes Responsible for the Production of the Cyclic Lipopeptide Arthrofactin by Pseudomonas sp MIS38
Kenji Washio, Siew Ping Lim, Niran Roongsawang, Masaaki Morikawa
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 74, 5, 992, 999, May 2010, [Peer-reviewed]
English, Scientific journal

The Role of Urease Activity on Biofilm Formation by Staphylococcus sp T-02 Isolated from the Toilet Bowl
Kaihei Oki, Kenji Washio, Daigo Matsui, Shinichi Kato, Yoshihiko Hirata, Masaaki Morikawa
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 74, 3, 583, 589, Mar. 2010, [Peer-reviewed]
English, Scientific journal

Flexible exportation mechanisms of arthrofactin in Pseudomonas sp MIS38
S. P. Lim, N. Roongsawang, K. Washio, M. Morikawa
JOURNAL OF APPLIED MICROBIOLOGY, 107, 1, 157, 166, Jul. 2009, [Peer-reviewed]
English, Scientific journal

Biofilm formation and proteolytic activities of Pseudoalteromonas bacteria that were isolated from fish farm sediments
Saori Iijima, Kenji Washio, Ryota Okahara, Masaaki Morikawa
MICROBIAL BIOTECHNOLOGY, 2, 3, 361, 369, May 2009, [Peer-reviewed]
English, Scientific journal

Production of sophorolipid biosurfactant by Pichia anomala
Jiraporn Thaniyavarn, Tanusta Chianguthai, Polakit Sangvanich, Niran Roongsawang, Keji Washio, Masaaki Morikawa, Suthep Thaniyavarn
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 72, 8, 2061, 2068, Aug. 2008, [Peer-reviewed]
English, Scientific journal

Identification of alkane hydroxylase genes in Rhodococcus sp strain TMP2 that degrades a branched alkane
Daisuke Takei, Kenji Washio, Masaaki Morikawa
BIOTECHNOLOGY LETTERS, 30, 8, 1447, 1452, Aug. 2008, [Peer-reviewed]
English, Scientific journal

Functional analysis of a pyoverdine synthetase from Pseudomonas sp MIS38
Slew Ping Lim, Niran Roongsawang, Kenji Washio, Masaaki Morikawa
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 71, 8, 2002, 2009, Aug. 2007, [Peer-reviewed]
English, Scientific journal

In vivo characterization of tandem C-terminal thioesterase domains in arthrofactin synthetase
Niran Roongsawang, Kenji Washio, Masaaki Morikawaa
CHEMBIOCHEM, 8, 5, 501, 512, Mar. 2007, [Peer-reviewed]
English, Scientific journal

Common mechanisms regulating expression of rice aleurone genes that contribute to the primary response for gibberellin
Kenji Washio, Masaaki Morikawa
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 1759, 10, 478, 490, Oct. 2006, [Peer-reviewed]
English, Scientific journal

Phylogenetic analysis of condensation domains in the nonribosomal peptide synthetases
N Roongsawang, SP Lim, K Washio, K Takano, S Kanaya, M Morikawa
FEMS MICROBIOLOGY LETTERS, 252, 1, 143, 151, Nov. 2005, [Peer-reviewed]
English, Scientific journal

Functional dissections between GAMYB and dof transcription factors suggest a role for protein-protein associations in the gibberellin-mediated expression of the RAmy1A gene in the rice aleurone
K Washio
PLANT PHYSIOLOGY, 133, 2, 850, 863, Oct. 2003, [Peer-reviewed]
English, Scientific journal

Cloning and characterization of cDNA of the GPI-anchored purple acid phosphatase and its root tissue distribution in Spirodela oligorrhiza
M Nishikoori, K Washio, A Hase, N Morita, H Okuyama
PHYSIOLOGIA PLANTARUM, 113, 2, 241, 248, Oct. 2001, [Peer-reviewed]
English, Scientific journal

Identification of Dof proteins with implication in the gibberellin-regulated expression of a peptidase gene following the germination of rice grains
K Washio
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 1520, 1, 54, 62, Jul. 2001, [Peer-reviewed]
English, Scientific journal

cDNA encoding Dof-proteins that are present in germinated aleurone cells (PGR 99.107).　　　　　　　　　　　　　　　
Washio K
Plant Physiology, 120, 1205, 1999, [Peer-reviewed]
English, Scientific journal

The glycosylphosphatidylinositol-anchored phosphatase from Spirodela oligorrhiza is a purple acid phosphatase
H Nakazato, T Okamoto, M Nishikoori, K Washio, N Morita, K Haraguchi, GA Thompson, H Okuyama
PLANT PHYSIOLOGY, 118, 3, 1015, 1020, Nov. 1998, [Peer-reviewed]
English, Scientific journal

ORGAN-SPECIFIC AND HORMONE-DEPENDENT EXPRESSION OF GENES FOR SERINE CARBOXYPEPTIDASES DURING DEVELOPMENT AND FOLLOWING GERMINATION OF RICE GRAINS
K WASHIO, K ISHIKAWA
PLANT PHYSIOLOGY, 105, 4, 1275, 1280, Aug. 1994, [Peer-reviewed]
English, Scientific journal

CLONING AND SEQUENCING OF THE GENE FOR TYPE-I CARBOXYPEPTIDASE IN RICE
K WASHIO, K ISHIKAWA
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 1199, 3, 311, 314, Apr. 1994, [Peer-reviewed]
English, Scientific journal

STRUCTURE AND EXPRESSION DURING THE GERMINATION OF RICE SEEDS OF THE GENE FOR A CARBOXYPEPTIDASE
K WASHIO, K ISHIKAWA
PLANT MOLECULAR BIOLOGY, 19, 4, 631, 640, Jul. 1992, [Peer-reviewed]
English, Scientific journal

QUANTITATIVE COMPARISON OF FERN-LEAF DEVELOPMENT AND FERTILITY WITH RESPECT TO ALTITUDE IN THE TIROL, CENTRAL EUROPEAN ALPS, AUSTRIA
T SATO, G GRABHERR, K WASHIO
JOURNAL OF BIOGEOGRAPHY, 16, 5, 449, 455, Sep. 1989, [Peer-reviewed]
English, Scientific journal

THERMOTROPIC PROPERTIES OF LIPIDS EXTRACTED FROM PLASMA-MEMBRANE AND TONOPLAST ISOLATED FROM CHILLING-SENSITIVE MUNG BEAN (VIGNA-RADIATA [L] WILCZEK)
S YOSHIDA, K WASHIO, J KENRICK, G ORR
PLANT AND CELL PHYSIOLOGY, 29, 8, 1411, 1416, Dec. 1988, [Peer-reviewed]
English, Scientific journal

紅藻マギレソゾLaurencia saitoi由来ブロモペルオキシダーゼの臭素化反応
正木志良, 金子賢介, 小林大毅, 石川高史, 西川慶祐, 森本善樹, 鷲尾健司, 森川正章, 沖野龍文, 日本農芸化学会大会講演要旨集(Web), 2016, 4B032 (WEB ONLY), 05 Mar. 2016
Japanese

紅藻ミツデソゾLaurencia okamurae由来ブロモペルオキシダーゼの性状解析
石川高史, 金子賢介, 湯暁蓉, 鷲尾健司, 森川正章, 沖野龍文, 化学系学協会北海道支部冬季研究発表会講演要旨集(CD-ROM), 2016, ROMBUNNO.1B17, 2016
Japanese

紅藻ソゾ属3種のブロモペルオキシダーゼの活性評価
金子賢介, 小林大毅, 鷲尾健司, 森川正章, 沖野龍文, 日本化学会講演予稿集, 95th, 4, 1409, 11 Mar. 2015
Japanese

紅藻ウラソゾLaurencia nipponica由来バナジウム依存型ブロモペルオキシダーゼの性状解析
KANEKO KENSUKE, WASHIO KENJI, UMEZAWA DAIKI, MATSUDA FUYUHIKO, MORIKAWA MASAAKI, OKINO TATSUFUMI, 日本水産学会大会講演要旨集, 2014, 119, 27 Mar. 2014
Japanese

The chromatin structure and seed function of land plants.　　　　　　　　　　　　　　　
Washio K, International Journal of Plant Biology & Research, 2, 1015, 2014, [Peer-reviewed], [Invited]
English, Introduction scientific journal

紅藻ウラソゾLaurencia nipponica由来バナジウム依存型ブロモペルオキシダーゼの性状解析
KANEKO KENSUKE, WASHIO KENJI, UMEZAWA DAIKI, MATSUDA FUYUHIKO, MORIKAWA MASAAKI, OKINO TATSUFUMI, マリンバイオテクノロジー学会大会講演要旨集, 15th, 68, 01 Jun. 2013
Japanese

The prediction of climate change and rice production in Japan.　　　　　　　　　　　　　　　
Washio K, Journal of Rice Research, 2, e103, 2013, [Peer-reviewed], [Invited]
English, Introduction scientific journal

Unique microbiological characteristics of Pseudoalteromonas spp. that form biofilms in the sea water environment : Their possible contribution to water purification and fish farming.　　　　　　　　　　　　　　　
Iijima S, Okahara R, Washio K, Morikawa M, Bulletin of the Society of Sea Water Science, 66, 186, 190, 2012, [Peer-reviewed]
Japanese, Introduction scientific journal

1Hp04 Mixed biofilm formation by Marinobacter bacteria and relatives
Morikawa Masaaki, Eiki Hisashi, Tarakita Natsumi, Washio Kenji, 日本生物工学会大会講演要旨集, 63, 54, 54, 2011
日本生物工学会, Japanese

Diversity of Nonribosomal Peptide Synthetases Involved in the Biosynthesis of Lipopeptide Biosurfactants
Niran Roongsawang, Kenji Washio, Masaaki Morikawa, INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 12, 1, 141, 172, Jan. 2011, [Peer-reviewed]
English, Book review

The role of urease activity on biofilm and urolith formation by Staphylococcus sp T-02 that was isolated from a toilet bowl
K. Oki, K. Washio, D. Matsui, Y. Hirata, M. Morikawa, JOURNAL OF BIOTECHNOLOGY, 150, S247, S247, Nov. 2010
English, Summary international conference

Pseudomonas stutzeriバイオフィルムのナフタレン分解活性と土壌安定性
嶋田恒平, 片岡剛文, 鷲尾健司, 森川正章, 日本農芸化学会大会講演要旨集, 2008, 39, 05 Mar. 2008
Japanese

S-C5 Sustainable bioremediation technology utilizing Biofilms
MORIKAWA Masaaki, WASHIO Kenji, 日本微生物生態学会講演要旨集, 24, 193, 193, 2008
日本微生物生態学会, Japanese

Duckweed-Rhizobacteria Symbiosis that Develops Sustainable Remediation Technology
YAMAGA Fumiko, WASHIO Kenji, MORIKAWA Masaaki, KAGAKU TO SEIBUTSU, 46, 10, 682, 688, 2008, [Invited]
微生物活性を利用した環境浄化法は，地球への負荷が少ない技術として広く認知されている．しかし，実用的技術としては，有用微生物の現場での定着率の低さによるコスト高や浄化後に危惧される微生物の異常増殖や生態系の攪乱などの問題が常に指摘されてきた．近年，この問題を解決する一つの方向性として根圏微生物の活用が注目されている．植物の光合成作用と微生物の汚染物質分解活性をリンクさせるこの手法の一例を紹介するとともに，今後の可能性を展望する．, Japan Society for Bioscience, Biotechnology, and Agrochemistry, Japanese, Introduction scientific journal

Analysis of transcription factors regulating gibberellin-inducible expression of a gene for a proteinase, Rep1 in rice seeds
Keita Sutoh, Ryozo Imai, Kenji Washio, Tomonori Nakai, Daisuke Yamauchi, PLANT AND CELL PHYSIOLOGY, 48, S191, S191, 2007
English, Summary international conference

Pseudomonas sutzeriバイオフィルムのナフタレン分解活性と土壌安定性
嶋田恒平, 片岡剛文, 鷲尾健司, 森川正章, 環境バイオテクノロジー学会大会プログラム講演要旨集, 31st, 14, 2007
Japanese

Expression of rice germination-control genes that accompany with the change of DNA modification.
K Washio, M Morikawa, PLANT AND CELL PHYSIOLOGY, 47, S147, S147, 2006
English, Summary international conference

枯草菌バイオフィルム形成遺伝子の探索
森川正章, 河合良尚, 相原 悠, 鷲尾健司, 高野和文, 金谷茂則, 生物工学会誌, 84, 12, 488, 490, 2006, [Invited]
日本生物工学会, Japanese, Introduction scientific journal

:Learning from the Microbial Activities which Exceed Central Dogma
WASHIO Kenji, MORIKAWA Masaaki, KAGAKU TO SEIBUTSU, 44, 2, 85, 92, 2006, [Invited]
Japan Society for Bioscience, Biotechnology, and Agrochemistry, Japanese, Introduction scientific journal

Dissection of DNA methylation status in gibberellin-primary response genes that involve the germination of rice seeds.
K Washio, M Morikawa, PLANT AND CELL PHYSIOLOGY, 46, S130, S130, 2005
English, Summary international conference

種子が目覚めるとき：発芽を促すジベレリンの作用　　　　　　　　　　　　　　　
鷲尾健司, 先端シンポジウム 植物科学の新展開 -分子から群集まで広視野研究をめざす- プログラム要旨集, 84, 92, 2004, [Invited]
Japanese, Summary national conference

RICE GRAIN DOF PROTEINS POSITIVELY REGULATE THE EXPRESSION OF GIBBERELLIN-RESPONSIVE GENES, COORDINATED WITH THE FUNCTION OF GAMYB :
WASHIO Kenji, Plant and cell physiology, 42, s34, 2001
Japanese Society of Plant Physiologists, English

CHARACTERIZATION OF DOF PROTEINS RELATED TO THE GIBBERELLIN REGULATED EXPRESSION OF GENES FOLLOWING THE GERMINATION OF RICE (Oryza sativa L.) GRAINS :
WASHIO Kenji, Plant and cell physiology, 41, s131, 2000
Japanese Society of Plant Physiologists, English

IDENTIFICATION OF LOW-MOLECULAR-WEIGHT DNA-BINDING PROTEINS THAT INTERACT WITH AN A/T-RICH PROMOTER SEQUENCE OF A GIBBERELLIN-INDUCIBLE GENE OF RICE (Oryza sativa)
WASHIO Kenji, Plant and cell physiology, 39, S50, S50, May 1998
English

PROMOTER ANALYSIS OF A GIBBERELLIN-INDUCIBLE GENE OF RICE (Oryza sativa) AND IDENTIFICATION OF ALEURONE PROTEINS THAT INTERACT WITH ITS 5'-POSITIVE REGURATORY REGION
WASHIO Kenji, Plant and cell physiology, 38, s137, Mar. 1997
English

EXPRESSION ANALYSIS OF A GENE FOR RICETYPE III CARBOXYPEPTIDASE USING OAT ALEURONE PROTOPLASTS
WASHIO Kenji, ISHIKAWA Koh, Plant and cell physiology, 36, S84, Mar. 1995
English

Biofilms in bioremediation, current research and emerging technologies: Using microbial biofilms to enhance the phytoremediation of contaminants in soil and water. Part A: A trial for sustainable phenol degradation by duckweed-colonizing biofilms.　　　　　　　　　　　　　　　
Morikawa M, Yamaga F, Suzuki K, Kurashina K, Miwa K, Washio K, Chapter 13, pp233–240
Caister Academic Press, 2015, [Joint work]

Biofilms in bioremediation, current research and emerging technologies: Comparison of the degradation activity of biofilm-associated versus planktonic cells.　　　　　　　　　　　　　　　
Morikawa M, Washio K, Chapter 12, pp221–232
Caister Academic Press, 2015, [Joint work]

バイオフィルムの基礎と制御 バイオサーファクタントを利用したバイオフィルムの形成と阻害　　　　　　　　　　　　　　　
鷲尾健司, 森川正章, pp278-287
(株)エヌ・ティー・エス, 2008, [Joint work]

レーヴン/ジョンソン 生物学 原著第7版　　　　　　　　　　　　　　　
鷲尾健司, 下巻、第36章、pp755-766
培風館, 2007, [Joint translation]

Plant nutrition-for sustainable food production and environment: Characterization and cDNA cloning of the GPI-anchored phosphatase from Spirodela oligorrhiza.　　　　　　　　　　　　　　　
Nakazato H, Okamoto T, Nishikoori M, Washio K, Morita N, Haraguchi K, Thompson GA Jr, Okuyama H, vol13, pp229-230
Kluwer Academic Publishers, Dordrecht, The Netherlands, 1997, [Joint work]

Study on bromination enzyme from the red alga Laurencia nipponica.　　　　　　　　　　　　　　　
Kaneko K, Washio K, Umezawa T, Morikawa M, Matsuda F, Okino T
Gordon Research Conferences, Marine Natural Products, 27 Feb. 2012, English, Nominated symposium
[Invited], [International presentation]

Staphylococcus属細菌のバイオフィルム形成における尿素分解酵素の役割　　　　　　　　　　　　　　　
大木海平, 鷲尾健司, 松井大悟, 平田善彦, 森川正章
平成22年度北海道腸内細菌叢研究会総会, 17 Sep. 2010, Japanese, Nominated symposium
[Invited], [Domestic Conference]

Generation of D-amino acid during biosynthesis of the cyclic lipopeptide arthrofactin by nonribosomal peptide synthetases.　　　　　　　　　　　　　　　
Washio K, Roongsawang N, Morikawa M
BIT 1st Annual World Congress of Catalytic Asymmetric Synthesis 2010, 20 May 2010, English, Nominated symposium
[Invited], [International presentation]

ウキクサと根圏細菌の相利共生作用による汚染浄化法　　　　　　　　　　　　　　　
森川正章, 鷲尾健司
水処理生物学会46回大会, 12 Nov. 2009, Japanese, Nominated symposium
[Invited], [Domestic Conference]

Biofilm formation and effective production of menaquinone-7 (Vitamin K2) by Bacillus subtilis.　　　　　　　　　　　　　　　
Washio K, Morikawa M
Japan-Argentina Workshop (JST/MYNCyT), 03 Aug. 2009, English, Public symposium
[Invited], [International presentation]

Genetic analysis of synthetic regulation and transportation of lipopeptide type biosurfactant.　　　　　　　　　　　　　　　
Morikawa M, Roongsawang N, Lim SP, Washio K
Biosurfactant Workshop in Vietnam, 27 Apr. 2009, English, Nominated symposium
[Invited], [International presentation]

Sustainable bioremediation technology by utilizing biofilms.　　　　　　　　　　　　　　　
Morikawa M, Yamaga F, Shimada K, Washio K
Int. Symposium on Biotechnological Approaches to Environmental Science for Energy Production and Sustainability, 23 Feb. 2009, English, Nominated symposium
[Invited], [International presentation]

Efficacy of biofilm formation by naphthalene degrading Pseudomonas stutzeri T102 toward bioremediation technology.　　　　　　　　　　　　　　　
Morikawa M, Shimada K, Washio K
BIT 1st World Congress of ibio-2008, 19 May 2008, English, Nominated symposium
[Invited], [International presentation]

環境微生物の生存戦略−バイオフィルム−　　　　　　　　　　　　　　　
森川正章, 相原 悠, 山崎和彦, 鷲尾健司
日本生化学会北海道支部 第44回支部例会, 06 Jul. 2007, Japanese, Keynote oral presentation
[Invited], [Domestic Conference]

種子が目覚めるとき 発芽を促すジベレリンの作用　　　　　　　　　　　　　　　
鷲尾健司
先端シンポジウム 植物科学の新展開 -分子から群集まで広視野研究をめざす-, 29 Oct. 2003, Japanese, Nominated symposium
[Invited], [Domestic Conference]

Studies on biosynthesis mechanisms responsible the production of marine halogenated compounds.　　　　　　　　　　　　　　　
WASHIO Kenji, 2010 - Present

Functional dissections of the adoptive growth in higher plants.　　　　　　　　　　　　　　　
WASHIO Kenji, 1994 - Present

Biosynthetic enzymes of antifouling halogenated compounds
Grants-in-Aid for Scientific Research
Apr. 2012 - Mar. 2016
Okino Tatsufumi, MATSUDA Fuyuhiko, UMEZAWA Taiki, NOGATA Yasuyuki, WASHIO Kenji
Vanadium dependent bromoperoxidase (VBPO) was cloned from the red alga Laurencia nipponica which produces a halogenated compound, laurencin. The enzyme catalyzed cyclization and bromination of a precursor of laurencin. Bromination activity of recombinant enzyme was investigated. A similar type of VBPO was cloned from L. okamurae which produces a halogenated sesquiterpenoid, laurinterol. The recombinant enzyme showed bromination activity. In addition, another type of VBPO was cloned from L. saitoi which produces a halogenated triterpenoid. Its properties were investigated by using its recombinant protein. The structure of bromoallene containing C15 compound, omaezallene was reported. Its totaly syntehsis was successfully achieved.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 24380112

Dissection of the genome functions that confer the dehydration tolerance to plant seeds.
Grants-in-Aid for Scientific Research
Apr. 2011 - Mar. 2014
WASHIO Kenji
Seed formation is a survival strategy of higher plants that protect the juvenile embryo from unfavorable conditions and facilitates the dispersion of offspring into a new habitat. The chromatin structures play important roles in a variety of nuclear functions by influencing the assembly of nuclear factors through epigenetic modifications. In the pioneer works of plant seed science, the relationships between structure and function of seed nuclei have been well documented, and the microscopic analyses have confirmed morphological changes of chromatin structures between the quiescent and germinated embryos, including the highly condensed state of chromatin structures. These observations demonstrate that the protective mechanisms of genome integrity must be mobilized in the plant seeds. In this study, we have purified the chromatin components from Arabidopsis dry seeds and conducted the subproteome analyzes by the use of 2D-PAGE in conjugation with mass spectrometric technology.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, Principal investigator, Competitive research funding, 23580468

Studies on the primitive alkane metabolisms and ecology of extremely thermophilic petroleum bacteria
Grants-in-Aid for Scientific Research
Apr. 2010 - Mar. 2014
MORIKAWA Masaaki, WASHIO Kenji
We determined whole genome sequence of Geobacillus thermoleovorans B23 and successfully discovered three sets of gene operons responsible for alkane degradation. We also indicated that the gene products are non-metal enzymes that should have been uniquely evolved under high temperature petroleum conditions. On the other hand, we have experimentally demonstrated that Coprothermobacter proteolyticus PM9-2, the first isolated bacterium from a deep subterranean high temperature oil reservoir, constituted syntrophy with a methanogenic archaeum. Moreover, it was found that they grew in coagulates that were covered by unique polysaccharides in order to exchange materials effectively. These results contribute to clarify microbial ecology in deep subterranean high temperature petroleum reservoir that is far isolated from surfaces.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, Competitive research funding, 22380175

Analyses on the molecular mechanisms of biofilm formation that reveal unknown functions of microorganisms
Grants-in-Aid for Scientific Research
Apr. 2007 - Mar. 2010
MORIKAWA Masaaki, WASHIO Kenji
Biofilms are highly structured community of surface attached microorganisms. Cell density in biofilms is so high enough to enable the member cells to communicate each other very effectively and express specific genes. Experimental results unveiled several unique characteristics of the biofilm cells and molecular mechanisms behind. Moreover, applying high stress-tolerance of the biofilm cells to pollutant degrading bacteria was successful in constructing base of sustainable environmental biotechnology.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, Competitive research funding, 19380189

Studies on molecular mechanisms that mobilize the expression of early GA-response genes in rice aleurone.
Grants-in-Aid for Scientific Research
Apr. 2004 - Mar. 2006
WASHIO Kenji
During germination of cereal seeds, aleurone cells respond to gibberellins (GA) by synthesizing and secreting hydrolytic enzymes that mobilize the reserved nutrients. It has been shown that products of early GA response genes, such as transcription factor GAMyb, act as key molecules leading to such regulation. Pivotal roles of GAMyb on the expression of hydrolase genes have been well documented, whereas regulation of GAMyb expression itself remains obscure. In order to understand virtual mechanisms of the GA-mediated expression of genes, it would be critical to know as to how GA control expression of early GA response genes. Using an aleurone transient expression system of rice (Oryza sativa L.), we examined GA responsive domains of GA primary genes in the aleurone, such as GAMyb and OsDof3. The 5' upstream portion could not confer GA responses. Extensive analyses revealed the presence of enhancer-like activities in a large first intron. In Arabidopsis, intron enhancers have been identified in MADS-box homeotic genes, AGAMOUS (AG) and FLOWERING LOCUS C (FLC), in which large introns should not only confer proper gene expressions, but also associate with gene silencing by covalent modifications of both DNA and histone. These evidences allow us to assign that chromatin-based control might be important for initial GA action. Based on this assumption, we have identified DNA methylation at GAMyb locus in germinated rice seeds. Bisulfite sequence analyses proposed that patterns of DNA methylation on the GAMyb promoter were possibly differing in the tissues or at hormonal conditions. These findings support an idea that chromatin configuration would be important for expression of early GA response genes and that GA likely permit target gene expressions by modulating chromatin architectures.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), HOKKAIDO UNIVERSITY, Principal investigator, Competitive research funding, 16570026

オオムギ糊粉層細胞表層におけるジベレリン酸受容体の検索
科学研究費補助金 奨励研究(A)
Apr. 1996 - Mar. 1997
鷲尾健司
澱粉性穀物植物はその種子発芽の際、植物ホルモンであるジベレリン酸(GA)を合成・分泌し貯蔵物質の分解利用に関与する加水分解酵素群の合成を誘導することが知られている。本研究ではこの種子発芽に関わる生理現象に着目し、植物ホルモンの作用機序を明らかにすることを目的とした。
GAの受容体はその作用組織である澱粉層細胞の原形質膜上に存在することが、微量注入法を用いた実験などにより示唆されている。そこで本研究ではオオムギ澱粉層細胞表層を認識する組換えファージ抗体発現ライブラリーを構築し、発現抗体とGA受容体が相互作用することにより示される生理活性の変化を指標としてGA受容体を免疫分子細胞学的に同定することを試みた。実験材料としてオオムギ澱粉層細胞より原形質膜画分を大量かつ高純度に単離することが必要であり、これには水性2層分配法とショ糖密度匂配分離法を併用することにより比較的純度の高い原形質膜画分を得ることができた。この原形質膜画分より調製したアジュバントをマウスに対して免疫操作を施し、常法に従いファージ抗体発現ライブラリーを作成した。このライブラリーより調製した一価性抗体は対照区に比べて著しくGAの効果を抑制することが確認されたため、GA受容体を認識する抗体を発現する組換えファージがライブラリー中に存在することが予想される。現在、このGA受容体を認識する抗体を発現する組換えファージを単離するための実験手法を模索している。
この実験と並行してGA応答性遺伝子の発見制御機構に関する研究も進めており、研究成果の一部は国際的学術雑誌であるMolecular Gene Genetics誌に受理され掲載準備中となっており、また日本植物生理学会は1997年度年会にて研究成果の概要を口頭発表した。
日本学術振興会, 奨励研究(A), 北海道大学, Principal investigator, Competitive research funding, 08780667

ジベレリン酸によるイネ・III型カルボキシペプチダーゼ遺伝子の発現制御の解析
科学研究費補助金 奨励研究(A)
Apr. 1994 - Mar. 1995
鷲尾健司
イネ・3型カルボキシペプチダーゼ(CPD3)遺伝子はイネ種子発芽時において糊粉層特異的に植物ホルモンであるジベレリン酸(GA)により発現誘導を受けるが、本研究ではこの遺伝子を用いて植物ホルモンによる遺伝子発現制御機構に関する知見を得ることを目的として実験を進めた。CPD3遺伝子より調整した各種プロモータ断片とGUS遺伝子を連結したレポーター遺伝子を用いて、オート麦・糊粉層より調整したプロトプラスト内での一過性の発現を見ることにより、GAにより発現誘導を受ける配列が転写開始点より上流90bpから273bpの間に存在することが確認された。この限定されたプロモーター断片に対するオート麦糊粉層核抽出物の結合状態をゲルシフト法を用いて解析した結果、183bpからなる配列内のA/T配列に富んだ領域に核内因子が結合していることが明らかになった。実際、このプロモーター配列内には非常にアデニン残基に富んだ領域が存在しているので、より詳細な核内因子の結合部位を同定するためフットプリント解析をおこなったところ、GA応答性遺伝子間に共通に見出されているピリミジンbox、TAACAGAboxを中心にして核内因子が結合していることが示唆された。GAの作用時、非作用時においてもゲルシフト解析においては同様にシフトしたバンドが検出されたが、フットプリント解析における核内因子の結合部位においては明確な違いを示した。GAの非作用時においてはピリミジンboxを中心にして核内因子が結合しており、GAの作用時にはTAACAGAboxにフットプリントが示された。特に、TAACAGAboxについては動物の癌原遺伝子産物であるMybのDNA結合配列(AACNG)を含んでいるので、ジベレリン酸の作用過程において植物のMyb相同物が重要な役割を担っている可能性が推定され、現在この点について詳細な研究を進めている。
日本学術振興会, 奨励研究(A), 北海道大学, Principal investigator, Competitive research funding, 06740589

新規水草根圏微生物　　　　　　　　　　　　　　　
Patent right, 森川正章, 鷲尾健司, 山賀文子
特願2008-099213, 07 Apr. 2008

2011 - 2011
新エネルギー・産業技術総合開発機構 受託研究
微生物機能を活用した環境調和型製造基盤技術開発 バイオフィルム工学による微生物のデザイン化

2009 - 2009
科学技術振興機構 戦略的国際科学技術協力推進事業
日本－アルゼンチン共催WS 農業・食糧生産に関するバイオサイエンス・バイオテクノロジー

2006 - 2006
日本学術振興会 拠点大学交流事業
微生物の生物化学的研究分野 耐熱性微生物資源の開発と利用

2006 - 2006
科学技術振興機構 サイエンスパートナーシッププログラム事業
講座型学習活動 生命と環境を科学する

2003 - 2003
科学技術振興機構 サイエンスパートナーシッププログラム事業
教育連携講座 北海道の農作物とバイオテクノロジー