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研究者情報

マスター

アカウント（マスター）

  • 氏名

    崎浜　靖子(サキハマ　ヤスコ), サキハマ　ヤスコ

所属（マスター）

  • 農学研究院　基盤研究部門　応用生命科学分野

所属（マスター）

  • 農学研究院　基盤研究部門　応用生命科学分野

独自項目

syllabus

  • 2021, 生体分子解析学特論, Advanced Analytical Biochemistry, 修士課程, 農学院, quantification, qualification, analytical method,
  • 2021, 応用生命科学概論, Introduction to Applied Biosciences, 学士課程, 農学部, 応用生命科学，植物育種学，遺伝子制御学，応用分子昆虫学，分子生物学，分子酵素学，生態化学生物学，分子環境生物学，生物情報分子解析学，ゲノム生化学
  • 2021, 化学Ⅱ, Chemistry II, 学士課程, 全学教育, 有機化合物、官能基、分子構造、化学的性質、化学反応、機能性有機物、生体関連有機物質
  • 2021, 化学実験, Laboratory Exercise of Chemistry, 学士課程, 農学部, 器具取扱、定量分析、定性分析、生理活性物質の合成・単離精製、抗生物質生産、植物二次代謝産物、スクリーニング
  • 2021, 生態化学, Ecological Chemistry, 学士課程, 農学部, 農薬／生理活性物質／植物生理
  • 2021, 一般教育演習(ﾌﾚｯｼｭﾏﾝｾﾐﾅｰ), Freshman Seminar, 学士課程, 全学教育, 天然物化学、生物有機化学、生理活性物質

researchmap

プロフィール情報

学位

  • 博士（理学）（琉球大学）

プロフィール情報

  • 姓

    崎浜, サキハマ

  • 名

    靖子, ヤスコ

  • ID各種

    B-1047-2012, 200901042177834648

対象リソース

業績リスト

研究キーワード

  • 活性酸素、活性窒素   フラボノイド   ベタレイン   植物色素   植物環境応答   Plant Physiology   

研究分野

  • ライフサイエンス / 食品科学 / 抗酸化
  • 環境・農学 / 植物保護科学 / 植物・微生物相互作用
  • ライフサイエンス / 食品科学 / 機能性植物二次代謝物
  • 環境・農学 / 植物保護科学 / 環境ストレス応答
  • ライフサイエンス / 植物分子、生理科学 / 環境適応

経歴

  • 2024年04月 - 現在 - 大学院農学研究院 教授
  • 2012年04月 - 2024年03月 - 大学院農学研究院 講師
  • 2007年04月 - 2012年03月 - 大学院農学研究院 助教
  • 2002年04月 - 2007年03月 - 大学院農学研究院 助手

受賞

  • 2017年12月 日本農芸化学会北海道支部 日本農芸化学会北海道支部奨励賞
     植物色素ベタレインの活性窒素消去機能 
    受賞者: 崎浜 靖子
  • 2004年02月 公益財団法人 井上科学振興財団 第20回 井上研究奨励賞
     高等植物におけるフラボノイドの抗酸化機能 
    受賞者: 崎浜 靖子

論文

  • Sopanat Sawatdee, Teeraya Jarunglumlert, Prasert Pavasant, Yasuko Sakihama, Adrian E. Flood, Chattip Prommuak
    BMC Plant Biology 23 1 2023年12月 [査読有り]
     
    Background: Light spectra have been demonstrated to result in different levels of comfort or stress, which affect plant growth and the availability of health-promoting compounds in ways that sometimes contradict one another. To determine the optimal light conditions, it is necessary to weigh the vegetable’s mass against the amount of nutrients it contains, as vegetables tend to grow poorly in environments where nutrient synthesis is optimal. This study investigates the effects of varying light conditions on the growth of red lettuce and its occurring nutrients in terms of productivities, which were determined by multiplying the total weight of the harvested vegetables by their nutrient content, particularly phenolics. Three different light-emitting diode (LED) spectral mixes, including blue, green, and red, which were all supplemented by white, denoted as BW, GW, and RW, respectively, as well as the standard white as the control, were equipped in grow tents with soilless cultivation systems for such purposes. Results: Results demonstrated that the biomass and fiber content did not differ substantially across treatments. This could be due to the use of a modest amount of broad-spectrum white LEDs, which could help retain the lettuce’s core qualities. However, the concentrations of total phenolics and antioxidant capacity in lettuce grown with the BW treatment were the highest (1.3 and 1.4-fold higher than those obtained from the control, respectively), with chlorogenic acid accumulation (8.4 ± 1.5 mg g− 1 DW) being particularly notable. Meanwhile, the study observed a high glutathione reductase (GR) activity in the plant achieved from the RW treatment, which in this study was deemed the poorest treatment in terms of phenolics accumulation. Conclusion: In this study, the BW treatment provided the most efficient mixed light spectrum to stimulate phenolics productivity in red lettuce without a significant detrimental effect on other key properties.
  • Yasuko Sakihama, Takashi Kato, Sopanat Sawatdee, Yo Yakushi, Junichi Asano, Hiroyo Hayashi, Yuya Goto, Makoto Hashimoto, Yasuyuki Hashidoko
    International Journal of Molecular Sciences 2023年10月 [査読有り][招待有り]
  • Wen Zhang, Kazu Sunami, Shuo Liu, Zihan Zhuang, Yasuko Sakihama, Da-Yang Zhou, Takeyuki Suzuki, Yuta Murai, Makoto Hashimoto, Yasuyuki Hashidoko
    Bioscience, Biotechnology, and Biochemistry 2023年08月01日 [査読有り]
     
    Abstract Squalene is a triterpenoid compound and widely used in various industries such as medicine and cosmetics due to its strong antioxidant and anticancer properties. The purpose of this study is to increase the accumulation of squalene in filamentous fungi using exogeneous butenafine hydrochloride, which is an inhibitor for squalene epoxidase. The detailed settings achieved that the filamentous fungi, Trichoderma virens PS1-7, produced squalene up to 429.93 ± 51.60 mg/L after culturing for 7 days in the medium consisted of potato infusion with glucose at pH 4.0, in the presence of 200 μM butenafine. On the other hand, no squalene accumulation was observed without butenafine. This result indicated that squalene was biosynthesized in the filamentous fungi PS1-7 which can be used as a novel source of squalene. In addition, we successfully obtained highly 13C-enriched squalene by using [U-13C6]-glucose as a carbon source replacing normal glucose.
  • Hitoshi Wakabayashi, Koji Sugiyama, Shinichi Suzuki, Yasuko Sakihama, Makoto Hashimoto, Martin J. Barwood
    European Journal of Applied Physiology 2022年10月28日 [査読有り]
  • Makoto Hashimoto, Manami Hashinoki, Natsumi Kurokawa, Yuta Murai, Zetryana Puteri Tachrim, Yasuko Sakihama, Takeyuki Suzuki
    HETEROCYCLES 103 1 392 - 392 2021年 [査読有り]
  • Masataka Hane, Hanny C. Wijaya, Yanetri A. Nyon, Yasuko Sakihama, Makoto Hashimoto, Hideyuki Matsuura, Yasuyuki Hashidoko
    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 85 1 77 - 84 2021年01月 [査読有り]
     
    Symbiosis of Penicillium rolfsii Y-1 is essential for the seed germination of Hawaii yellow-eyed grass (Xyris complanata). However, the local soil where the plants grow naturally often suppresses the radicle growth of the seedlings. This radicle growth was drastically restored by coinoculation of Paraburkholderia phenazinium isolate CK-PC1, which is a rhizobacterium of X. complanata. It was found that the isolate CK-PC1 produced phenazine-1-carboxylic acid (PCA, 1) as a major metabolite. The biological effects of PCA (1) were investigated using the seeds of X. complanata and Mung bean (Vigna radiata) and it was uncovered that the symbiosis of the isolate CK-PC1was essential for the postgermination growth of X. complanata and the metabolite PCA (1) might partially contribute to promote the growth of the plants.
  • Tachrim, Zetryana Puteri, Nakagawa, Shiori, Nakamura, Tadashi, Ohashi, Fumina, Kurokawa, Natsumi, Wakasa, Haruna, Tokoro, Yurika, Sakihama, Yasuko, Hashidoko, Yasuyuki, Suzuki, Takeyuki, Hashimoto, Makoto
    HETEROCYCLES 99 1 404 - 414 2019年04月 [査読有り][通常論文]
     
    CycloDOPA (5,6-dihydroxy-indoline-2-carboxylic acid, leukodopachrome) is one of metabolites derived from tyrosine, one of intermediate in melanin formation (mammalian) and betanidin main skeleton (betalain pigment in plant). Synthesis of deuterated cyclodopa via hydrogen/deuterium exchange by utilization of deuterium chloride (DCl) and deuterated triflic acid (TfOD) are reported. The novel fully deuterated aromatic cycloDOPA derivative can be formed depending on temperature and time of H/D exchange condition. The complete study of H/D exchange resulted in the selective deuterium between 4- and/or 7-position of aromatic hydrogen of cycloDOPA.
  • Kurokawa, Natsumi, Tokoro, Yurika, Tachrim, Zetryana Puteri, Wakasa, Haruna, Sakihama, Yasuko, Hashidoko, Yasuyuki, Hashimoto, Makoto
    ARKIVOC 42 - 49 2019年 [査読有り][通常論文]
     
    A chiral N-protected alpha-amino acid N-hydroxysuccinimide ester (OSu) is a common useful reagent for peptide bond formation. Recently, N-trifluoroacetyl (TFA) alpha-amino acid OSu esters have been reported as acyl donors for Frieldel-Crafts reactions to synthesize chiral alpha-amino phenyl ketones retaining the configurations of the starting alpha-amino acids. There are few reports for chiral TFA protected methionine, which has methylthioethyl structure in the side-chain of the alpha-amino acid. The detailed synthesis of chiral TFA-Met-OSu and its application as an acyl donor for Friedel-Crafts acylation is reported.[GRAPHICS].
  • Tachrim, Zetryana Puteri, Oida, Kazuhiro, Ohashi, Fumina, Wakasa, Haruna, Ikemoto, Haruka, Kurokawa, Natsumi, Sakihama, Yasuko, Hashidoko, Yasuyuki, Suzuki, Takeyuki, Hashimoto, Makoto
    HETEROCYCLES 97 2 877 - 893 2018年09月 [査読有り][通常論文]
     
    The utilization of N-trifluoroacetyl (TFA)-alpha-amino acid N-hydroxysuccinimide ester (OSu) derivatives, a promising acylating agent with high storage stability, is reported for Friedel Crafts acylation into arenes and N-heterocycles. The reaction between TFA-Phe-OSu derivatives and arenes afforded inter- and intramolecular products. TFA-Tyr-OSu derivatives, which possess hydroxyl substituent in the aromatic moiety of phenylalanine, afforded only intermolecular product with benzene. The heterocyclic TFA-Pro-OSu also shows relatively high reactivity toward acylation.
  • Shiori Nakagawa, Zetryana Puteri Tachrim, Natsumi Kurokawa, Fumina Ohashi, Yasuko Sakihama, Takeyuki Suzuki, Yasuyuki Hashidoko, Makoto Hashimoto
    Molecules 23 8 1943  2018年 [査読有り][通常論文]
     
    © 2018 by the authors. CycloDOPA (leukodopachrome), a well known metabolite of tyrosine, is a precursor of melanine in mammalian organisms and of the pigment betalain in plants. However, the isolation of cycloDOPA from natural sources has not been widely reported. In the present work, the stabilities of cycloDOPA and cycloDOPA methyl ester at various pH levels were studied. Both compounds were stable under acidic conditions. By contrast, both compounds were unstable when the pH was shifted from neutral to basic to form indole derivatives as major products. Based on the pH stability, cycloDOPA and its derivatives were subjected to the DPPH radical scavenging assay for the first time.
  • Lei Wang, Zetryana Puteri Tachrim, Natsumi Kurokawa, Fumina Ohashi, Haruna Wakasa, Yasuko Sakihama, Yasuyuki Hashidoko, Takeyuki Suzuki, Makoto Hashimoto
    ARKIVOC 58 - 65 2018年 [査読有り][通常論文]
     
    Sucrose is well known as naturally occurring sweeteners. Photoreactive sucrose derivative containing 3-(trifluoromethyl)diazirinyl moiety is designed for photoaffinity labeling. As 1'-hydroxyl group of sucrose is well known to be less reactive than other primary alcohols, the optimization of reaction conditions for diazirinyl benzyl bromide derivative at sucrose 1'-portion was examined to elucidate the functional analysis of sweet receptors.[GRAPHICS].
  • Tachrim, Zetryana Puteri, Nakamura, Tadashi, Sakihama, Yasuko, Hashidoko, Yasuyuki, Hashimoto, Makoto
    ARKIVOC 341 - 348 2018年 [査読有り][通常論文]
     
    1-kestose (O-B-D-fructofuranosyl-(2 -> 1)-B-D-fructofuranosyl-(2 -> 1)-alpha-D-glucopyranoside) is a potential short chain fructooligosaccharide with an inulin-type skeleton. Halogenation of 1-kestose was conducted via the Appel reaction with the use of carbon tetrahalide (CBr4 or CCI4) and triphenylphosphine, which was then followed by conventional acetylation. The per-O-acetylated form of 6,6',6 ''-trihalogenated derivatives of 1-kestose were conveniently isolated. Further deprotection of the per-O-acetylated form resulted in 6-, 6'-, and 6 ''-trihalogenated derivatives. The structure elucidation by one- and two-dimensional nuclear magnetic resonance established that halogenations are specific at the 6-, 6'-, and 6 ''-position of 1-kestose primary alcohols.[GRAPHICS].
  • Dian Anggraini Suroto, Shigeru Kitani, Kiyoko T. Miyamoto, Yasuko Sakihama, Masayoshi Arai, Haruo Ikeda, Takuya Nihira
    JOURNAL OF BIOSCIENCE AND BIOENGINEERING 124 6 611 - 617 2017年12月 [査読有り][通常論文]
     
    The genomes of actinomycetes encode many cryptic novel/useful bioactive compounds, but access to these cryptic secondary metabolites remains limited. Streptomyces avermitilis predominantly produces three polyketide antibiotics (avermectin, filipin, and oligomycin) but has the potential to produce more secondary metabolites based on the number of cryptic biosynthetic gene clusters. Here, we extensively investigated the metabolite profiles of a gene disruptant of AvaR3 (an autoregulator receptor homologue), which is involved in the pleiotropic regulation of antibiotic production and cell morphology. Unlike the wild-type strain, the avaR3 mutant accumulated compound 3 in the culture. The chemical structure of compound 3 was elucidated on the basis of various spectroscopic analyses, and was identified as phthoxazolin A, a cellulose synthesis inhibitor. Bioassays demonstrated that compound 3 exerts growth inhibitory activity against a broad range of plant pathogenic oomycetes. Moreover, unlike avermectin production, phthoxazolin A (3) production was negatively controlled by avenolide, a new type of autoregulator in streptomycetes, through the function of AvaR3. These results suggest that the genetic manipulation of autoregulator receptor homologues would be a valuable tool for the discovery of cryptic bioactive compounds. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.
  • Tachrim, Zetryana Puteri, Oida, Kazuhiro, Ikemoto, Haruka, Ohashi, Fumina, Kurokawa, Natsumi, Hayashi, Kento, Shikanai, Mami, Sakihama, Yasuko, Hashidoko, Yasuyuki, Hashimoto, Makoto
    MOLECULES 22 10 2017年10月 [査読有り][通常論文]
     
    Chiral N-protected alpha-amino aryl-ketones are one of the useful precursors used in the synthesis of various biologically active compounds and can be constructed via Friedel-Crafts acylation of N-protected alpha-amino acids. One of the drawbacks of this reaction is the utilization of toxic, corrosive and moisture-sensitive acylating reagents. In peptide construction via amide bond formation, N-hydroxysuccinimide ester (OSu), which has high storage stability, can react rapidly with amino components and produces fewer side reactions, including racemization. This study reports the first synthesis and utilization of N-trifluoroacetyl (TFA)-protected alpha-amino acid-OSu as a potential acyl donor for Friedel-Crafts acylation into various arenes. The TFA-protected isoleucine derivative and its diastereomer TFA-protected allo-isoleucine derivative were investigated to check the retention of alpha-proton chirality in the Friedel-Crafts reaction. Further utilization of OSu in other branched-chain and unbranched-chain amino acids results in an adequate yield of TFA-protected alpha-amino aryl-ketone without loss of optical purity.
  • Lei Wang, Zetryana Puteri Tachrim, Natsumi Kurokawa, Fumina Ohashi, Yasuko Sakihama, Yasuyuki Hashidoko, Makoto Hashimoto
    MOLECULES 22 8 2017年08月 [査読有り][通常論文]
     
    Aliphatic diazirines have been widely used as prominent photophores for photoaffinity labeling owing to their relatively small size which can reduce the steric effect on the natural interaction between ligands and proteins. Based on our continuous efforts to develop efficient methods for the synthesis of aliphatic diazirines, we present here a comprehensive study about base-mediated one-pot synthesis of aliphatic diazirines. It was found that potassium hydroxide (KOH) can also promote the construction of aliphatic diazirine with good efficiency. Importantly, KOH is cheaper, highly available, and easily handled and stored compared with the previously used base, potassium tert-butoxide (t-BuOK). Gram-scale study showed that it owned great advantages in being used for the large-scale production of aliphatic diazirines. This protocol is highly neat and the desired products can be easily isolated and purified. As the first comprehensive study of the base-mediated one-pot synthesis of aliphatic diazirines, this work provided good insight into the preparation and utilization of diazirine-based photoaffinity labeling probes.
  • Makoto Hashimoto, Takuma Yoshida, Zetryana Puteri Tachrim, Yasuko Sakihama, Yasuyuki Hashidoko, Yasumaru Hatanaka, Yuichi Kanaoka
    HETEROCYCLES 95 1 462 - 473 2017年01月 [査読有り][通常論文]
     
    O-Benzylation of serine is one of the important protection methods for solid phase peptide synthesis. The utilities of the protection group may be indicated that chemical modifications for O-benzylserine will be utilized to make functional peptides on solid phase synthesis. Detailed studies for effective synthesis of photoreactive and fluorophore containing O-benzylserine derivatives without racemization were reported.
  • Akiko Ishida, Lei Wang, Zetryana Puteri Tachrim, Takeyuki Suzuki, Yasuko Sakihama, Yasuyuki Hashidoko, Makoto Hashimoto
    CHEMISTRYSELECT 2 1 160 - 164 2017年01月 [査読有り][通常論文]
     
    Phenylthiourea (PTU) is well known as bioactive compound and one of the reversible inhibitors for tyrosinase. Tyrosinase plays very important roles for tyrosine metabolisms to maintain the skin homeostasis from sunlight with forming melanin. Synthesis of photoaffinity label reagents of PTU will be attractive thesis to elucidate functional analysis for tyrosinase.
  • Yudai Yamashita, Maremichi Ota, Yutaka Inoue, Youko Hasebe, Masanori Okamoto, Tsuyoshi Inukai, Chikra Masuta, Yasuko Sakihama, Yasuyuki Hashidoko, Mikiko Kojima, Hitoshi Sakakibara, Yasuyuki Inage, Kosaku Takahashi, Teruhiko Yoshihara, Hideyuki Matsuura
    PLANT AND CELL PHYSIOLOGY 57 5 986 - 999 2016年05月 [査読有り][通常論文]
     
    Plant hormones are a group of structurally diverse small compounds that orchestrate the cellular processes governing proper plant growth and environmental adaptation. To understand the details of hormonal activity, we must study not only their inherent activities but also the cross-talk among plant hormones. In addition to their use in agriculture, plant chemical activators, such as probenazole and uniconazole, have made great contributions to understand hormonal cross-talk. However, the use of plant chemical activators is limited due to the lack of activators for certain hormones. For example, to the best of our knowledge, there are only a few chemical activators previously known to stimulate the accumulation of ABA in plants, such as absinazoles and proanthocyanidins. In many cases, antagonistic effects have been examined in experiments using exogenously applied ABA, although these studies did not account for biologically relevant concentrations. In this report, it was found that a natural product, theobroxide, had potential as a plant chemical activator for stimulating the accumulation of ABA. Using theobroxide, the antagonistic effect of ABA against GAs was proved without exogenously applying ABA or using mutant plants. Our results suggest that ABA levels could be chemically controlled to elicit ABA-dependent biological phenomena.
  • Lei Wang, Takuma Yoshida, Yasuyuki Muto, Yuta Murai, Zetryana Puteri Tachrim, Akiko Ishida, Shiori Nakagawa, Yasuko Sakihama, Yasuyuki Hashidoko, Katsuyoshi Masuda, Yasumaru Hatanaka, Makoto Hashimoto
    EUROPEAN JOURNAL OF ORGANIC CHEMISTRY 14 3129 - 3134 2015年05月 [査読有り][通常論文]
     
    Saccharin is one of the most common artificial sweeteners that has a bitter taste at high concentrations. Currently, there are no detailed functional analyses of these gustatory receptors. Therefore, we designed and synthesized photoreactive saccharin derivatives that contain a (trifluoromethyl) diazirinyl moiety at the 5- or 6-position for use as functional analysis tools for photoaffinity labeling.
  • Lei Wang, Yuta Murai, Takuma Yoshida, Akiko Ishida, Katsuyoshi Masuda, Yasuko Sakihama, Yasuyuki Hashidoko, Yasumaru Hatanaka, Makoto Hashimoto
    ORGANIC LETTERS 17 3 616 - 619 2015年02月 [査読有り][通常論文]
     
    Alternative one-pot synthesis of 3-(trifluoromethyl)-3-phenyldiazirine derivatives from corresponding tosyloximes is developed. The deprotonation of intermediate diaziridine by NH2- is a new approach for construction of diazirine. Moreover, a novel synthesis of optically pure (trifluoromethyl)diazirinylphenylalanine derivatives was attempted involving these methods.
  • Munenori Sakurai, Takuma Yoshida, Lei Wang, Yuta Murai, Katsuyoshi Masuda, Yasuko Sakihama, Yasuyuki Hashidoko, Yasumaru Hatanaka, Makoto Hashimoto
    HETEROCYCLES 90 1 698 - 705 2015年01月 [査読有り][通常論文]
     
    Salicin (salicyl alcohol glucoside) is a substance well known for its bitter taste. A photoreactive diazirinyl derivative of salicin will be utilized for the functional analysis of interactions between the bitter taste receptor and salicin. Glucosides of salicyl derivatives are more difficult than phenol derivatives that are unsubstituted at the ortho-position. A diazirinyl salicin derivative was synthesized at moderate yields by glucosidation of 2,3,4,6-tetra-O-acetyl-alpha-D-glucopyranosyl bromide and 2-hydroxy-4-[3-(trifluoromethyl)-3H-diazirin-3-yl]benzaldehyde in the presence of a phase-transfer catalyst, nBuEt(3)NBr, followed by reduction and deprotection.
  • Takuma Yoshida, Lei Wang, Shiori Nakagawa, Zetryana Puteri Tachrim, Yasuko Sakihama, Yasuyuki Hashidoko, Makoto Hashimoto
    LETTERS IN ORGANIC CHEMISTRY 12 8 544 - 548 2015年 [査読有り][通常論文]
     
    Synthesis of cross-linkable diketopiperadine derivatives is described. Cross-linkable a-amino acid methyl esters were subjected to peptide synthesis with Boc-protected glycine or L-tyrosine. No protection of cross-linkable functional groups (catechol and phenylazide) is necessary for the construction of diketopiperazine skeleton.
  • Lei Wang, Yuta Murai, Takuma Yoshida, Masashi Okamoto, Katsuyoshi Masuda, Yasuko Sakihama, Yasuyuki Hashidoko, Yasumaru Hatanaka, Makoto Hashimoto
    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 78 7 1129 - 1134 2014年07月 [査読有り][通常論文]
     
    In this paper we report here a hydrogen/deuterium exchange (H/D exchange) of cross-linkable alpha-amino acid derivatives with deuterated trifluoromethanesulfonic acid (TfOD). H/D exchange with TfOD was easily applied to o-catechol containing phenylalanine (DOPA) within an hour. A partial H/D exchange was observed for trifluoromethyldiazirinyl (TFMD) phenylalanine derivatives. N-Acetyl-protected natural aromatic alpha-amino acids (Tyr and Trp) were more effective in H/D exchange than unprotected ones. The N-acetylated TFMD phenylalanine derivative afforded slightly higher H/D exchange than unprotected derivatives. An effective post-deuteration method for cross-linkable alpha-amino acid derivatives will be useful for the analysis of biological functions of bioactive peptides and proteins by mass spectrometry.
  • Sastia Prama Putri, Kei-ichi Ishido, Hiroshi Kinoshita, Shigeru Kitani, Fumio Ihara, Yasuko Sakihama, Yasuhiro Igarashi, Takuya Nihira
    JOURNAL OF BIOSCIENCE AND BIOENGINEERING 117 5 557 - 562 2014年05月 [査読有り][通常論文]
     
    A total of 412 strains belonging to 14 genera of clavicipitoid entomopathogenic fungi (EPF) were screened for activities against two economically important plant pathogenic oomycetes, Phytophthora sojae and Aphanomyces cochlioides. To identify the antioomycete compounds produced by EPF, the extracts of 13 highly active EPF strains were characterized in detail by high performance liquid chromatography with diode array detection and high-resolution mass spectrometric detection and antioomycete assay. The antioomycete activity of several Metarhizium extracts was associated with previously isolated aurovertins, fungerin, N-(methyl-3-oxodec-6-enoyl)-2-pyrroline, and N-(methyl-3-oxodecanoyl)-2-pyrroline. The depsipeptide beauvericin was confirmed to be one of the active principles of three strains of Isaria tenuipes, which strongly inhibited mycelial growth of both P. sojae and A. cochlioides. Two known bioactive metabolites, paecilosetin and aranorosinol A, together with a novel and potent antioomycete compound, farinomalein, were isolated from the extracts of Isaria farinosa and all compounds were confirmed to have antioomycete activity. Identification of 8 antioomycete compounds from 13 clavicipitioid EPF demonstrated a new potential use of EPF as a source of compounds for the control of soil-borne plant pathogenic oomycetes. (C) 2013, The Society for Biotechnology, Japan. All rights reserved.
  • Lei Wang, Takuma Yoshida, Masashi Okamoto, Yasuko Sakihama, Yasuyuki Hashidoko, Makoto Hashimoto
    MolBank 2014 1 M816 - M816 2014年02月24日 [査読有り][通常論文]
     
    The title compound, which has two photoreactive groups in a molecule, was synthesized by the coupling reaction of 3-(4-(bromomethyl)phenyl)-3-(trifluoromethyl)- 3H-diazirine in the presence of silver oxide in DMSO. © 2014 by the authors licensee MDPI, Basel, Switzerland.
  • Sakurai Munenori, Masuda Katsuyoshi, Wang Lei, Murai Yuta, Sakihama Yasuko, Hashidoko Yasuyuki, Hatanaka Yasumaru, Hashimoto Makoto
    HETEROCYCLES 88 1 629 - 637 2014年01月 [査読有り][通常論文]
     
    Photoreactive phenylalanine derivatives are well known as functional analysis reagents for target biomolecules. The photophores are commonly introduced at 4-position on benzene. Aspartame, which consists of dipeptide L-Asp-L-Phe-OMe, is one of the most utilized artificial sweeteners, and substitution effects on its benzene ring have been reported. Substitution at the 4-position, however, does not maintain its sweetness properties. Trifluoromethyldiazirine, which is one of the most reliable photophores, was introduced to a different site on phenylalanine and the new photoreactive phenylalanine was converted to aspartame derivatives. The new aspartame derivative had slightly higher sweetness potency than sucrose standard solution.
  • Yuta Murai, Lei Wang, Yasuyuki Muto, Yasuko Sakihama, Yasuyuki Hashidoko, Yasumaru Hatanaka, Makoto Hashimoto
    HETEROCYCLES 87 10 2119 - 2126 2013年10月 [査読有り][通常論文]
     
    Simple and stereocontrolled preparation of benzoylated phenylalanine derivatives from optically pure phenylalanine using Friedel-Crafts reaction in trifluoromethanesulfonic acid (TfOH) is reported; these derivatives are useful for photoaffinity labeling. Protected or unprotected phenylalanine derivatives were converted to benzoyl derivatives in TfOH at room temperature in a short time without loss of optical purity. The reaction condition was applied to synthesize novel photoreactive phenylalanine derivative, which has two photophores (benzophenone and diazirine). The detail analysis of photo-irradiation for two different photophores contained phenylalanine derivative was also investigated.
  • Lei Wang, Wataru Hisano, Yuta Murai, Munenori Sakurai, Yasuyuki Muto, Haruka Ikemoto, Masashi Okamoto, Takashi Murotani, Reika Isoda, Dongyeop Kim, Yasuko Sakihama, Irnayuli R. Sitepu, Yasuyuki Hashidoko, Yasumaru Hatanaka, Makoto Hashimoto
    MOLECULES 18 7 8393 - 8401 2013年07月 [査読有り][通常論文]
     
    Photoaffinity labeling is a reliable analytical method for biological functional analysis. Three major photophores-aryl azide, benzophenone and trifluoromethyldiazirine-are utilized in analysis. Photophore-bearing L-phenylalanine derivatives, which are used for biological functional analysis, were inoculated into a Klebsiella sp. isolated from the rhizosphere of a wild dipterocarp sapling in Central Kalimantan, Indonesia, under nitrogen-limiting conditions. The proportions of metabolites were quite distinct for each photophore. These results indicated that photophores affected substrate recognition in rhizobacterial metabolic pathways, and differential photoaffinity labeling could be achieved using different photophore-containing L-phenylalanine derivatives.
  • Mengcen Wang, Tomohiko Takayama, Dongyeop Kim, Yasuko Sakihama, Satoshi Tahara, Yasuyuki Hashidoko
    JOURNAL OF PESTICIDE SCIENCE 38 3-4 181 - 187 2013年 [査読有り][通常論文]
     
    Cochliophilin A (5-hydroxy-6,7-methylenedioxyflavone, 1) and N-(E)-feruloy1-4-0-methyldopamine (2) are naturally occurring host-specific chemoattractants for Aphanomyces cochlioides zoospores. In a cross-competition assay, compound 1 (4 fmol) applied to diatomite particles clearly attracted A. cochlioides zoospores in an aqueous solution of excessive compound 2 (lx 10(-6)M) that could mask a concentration gradient of 1 dispersed from the particles. Similarly, 2 (40 fmol) on particles also attracted A. cochlioides zoospores in an aqueous solution of 1 (1 X 10(-6)M). In addition, compound 2 on the particles did not attract the zoospores in an aqueous solution containing 1 X10(-6)M NADP(+), while 1 clearly attracted zoospores in the same solution. These results allowed us to speculate that compounds 1 and 2 do not share receptors. The chemosensory system for 1 is probably for host recognition of A. cochlioides zoospores, while the system for 2 is linked to cell differentiation. (C) Pesticide Science Society of Japan
  • Yuta Murai, Lei Wang, Katsuyoshi Masuda, Yasuko Sakihama, Yasuyuki Hashidoko, Yasumaru Hatanaka, Makoto Hashimoto
    European Journal of Organic Chemistry 2013 23 5111 - 5116 2013年 [査読有り][通常論文]
     
    Novel hydrogen/deuterium exchange for aromatic α-amino acids and their corresponding peptides were performed through the use of deuterated trifluoromethanesulfonic acid (TfOD). Detailed analysis of the exchange revealed that equal hydrogen/deuterium exchange was observed for phenylalanine, and specific exchange at the ortho-positions of phenol for tyrosine was also detected. The stereochemistry of the aromatic α-amino acids was retained under the exchange conditions. The hydrogen/deuterium exchange properties for these aromatic α-amino acids are identical for peptides that contain several aromatic α-amino acids. The exchange proceeded significantly faster than previous methods. Detailed analysis of the exchange revealed that the method could be controlled by temperature, time, and the amount of reagent. Deuterated trifluoromethanesulfonic acid promotes rapid and controllable hydrogen/deuterium exchange on aromatic rings of α-amino acids and peptides at room temperature and below. The deuterium incorporation depends on the amount of reagent, the temperature, and the time-scale of the reaction. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
  • Yui Ogasawara, Yuta Murai, Yasuko Sakihama, Yasuyuki Hashidoko, Makoto Hashimoto
    HETEROCYCLES 86 1 735 - 743 2012年12月 [査読有り][通常論文]
     
    The click reaction is one of the latest techniques for the chemical modification of bioactive compounds. Chemical modifications of alpha-amino acid side chains are gaining significance as useful and important tools for biochemical research. Biotinylation at side chain of propargylglycine using click reaction was examined. The detail quantitative analysis of Cu(I) concentration are performed to proceed the click reaction effectively.
  • Yui Ogasawara, Yuta Murai, Yasuko Sakihama, Yasuyuki Hashidoko, Makoto Hashimoto
    International Journal of Organic Chemistry 2 3 302 - 304 2012年11月 [査読有り][通常論文]
  • Yasuyuki Muto, Yuta Murai, Yasuko Sakihama, Yasuyuki Hashidoko, Makoto Hashimoto
    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 76 11 2162 - 2164 2012年11月 [査読有り][通常論文]
     
    Biotin is one of the most useful tags in (bio)analytical science due to its specific interaction with avidin, but is not easy to convert because of its low solubility in most solvents. Friedel-Crafts acylation of biotin acid chloride in triflic acid was examined, and the synthesized derivatives had stronger affinity to avidin than biotin in a binding assay using 2-(4'-hydroxyphenylazo)benzoic acid.
  • Yuta Murai, Katsuyoshi Masuda, Yasuko Sakihama, Yasuyuki Hashidoko, Yasumaru Hatanaka, Makoto Hashimoto
    JOURNAL OF ORGANIC CHEMISTRY 77 19 8581 - 8587 2012年10月 [査読有り][通常論文]
     
    5- and 6-trifluoromethyldiazirinyl indoles were synthesized from corresponding bromoindole derivatives for the first time. They acted as mother skeletons for the comprehensive synthesis of various bioactive indole metabolites. These can be used in biological functional analysis as diazirine-based photoaffinity labels.
  • Yasuko Sakihama, Makiko Maeda, Makoto Hashimoto, Satoshi Tahara, Yasuyuki Hashidoko
    FREE RADICAL RESEARCH 46 1 93 - 99 2012年01月 [査読有り][通常論文]
     
    Two major betalains, red-purple betacyanins and yellow betaxanthins, were isolated from red beetroots (Beta vulgaris L.), and their peroxynitrite (ONOO(-)) scavenging capacity was investigated. Apparent colours of the betalains were bleached by the addition of ONOO(-), and the absorbance decreases were suppressed in the presence of glutathione, a ONOO(-) scavenger. After bleaching, a new absorption maximum was observed at 350 nm in the spectrum of the resulting reaction mixture. New peaks were detected from HPLC analysis of the reaction products of betanin, a representative constituent of red beetroot betacyanins, treated with ONOO(-) monitoring at 350 nm, and the intensity of the major peak was positively correlated with ONOO(-) concentration. Betanin inhibited the ONOO(-) (0.5 mM)-dependent nitration of tyrosine (0.1 mM). Additionally, the IC(50) value of betanin (19.2 mu M) was lower than that of ascorbate (79.6 mu M). The presence of betanin (0.05-1.0 mM) also inhibited ONOO(-) (0.5 mM)-dependent DNA strand cleavage in a concentration -dependent manner. These results suggest that betalains can protect cells from nitrosative stress in addition to protecting them from oxidative stresses.
  • Md. Tofazzal Islam, Mitsuyoshi Sakasai, Yasuyuki Hashidoko, Abhinandan Deora, Yasuko Sakihama, Satoshi Tahara
    Journal of General Plant Pathology 73 5 324 - 329 2007年10月 [査読有り][通常論文]
     
    A modified medium was used to culture mycelium and produce a large quantity of zoospores of Aphanomyces cochlioides, a principal pathogen of damping-off disease of sugar beet and root rot disease of spinach. The semisolid medium consisted of 17 g corn meal agar (CMA) added with 4 g of yeast extract (YE) per liter of 50 mM phosphate buffer (pH 6.8-7.0). This medium supported the production of ca. 106 zoospores ml-1 in 6-day-old cultures, approximately 11-fold higher than the commonly used CMA (17 g CMA per liter of water, pH 6.0 ± 0.2). Although morphological characters of the zoospores produced from the hyphae grown on CMA and CMA + YE were almost similar, they contrasted their developmental strategy after encystment induced by mechanical agitation. Cystospores originating from the zoospores on CMA regenerated zoospores (> 80%), while those from CMA + YE germinated (ca. 80%) and produced hyphae. Furthermore, 4-10% of the germinated cystospores on CMA + YE had double germ tubes. The soluble protein profiles of zoospores produced on CMA and on CMA + YE demonstrated that several proteins were either different or expressed differently. Our results suggest that the culture medium directly influences zoosporogenesis in A. cochlioides hyphae and the developmental strategy of the produced zoospores. © 2007 The Phytopathological Society of Japan and Springer.
  • Brett M. Tyler, Sucheta Tripathy, Xuemin Zhang, Paramvir Dehal, Rays H. Y. Jiang, Andrea Aerts, Felipe D. Arredondo, Laura Baxter, Douda Bensasson, Jim L. Beynon, Jarrod Chapman, Cynthia M. B. Damasceno, Anne E. Dorrance, Daolong Dou, Allan W. Dickerman, Inna L. Dubchak, Matteo Garbelotto, Mark Gijzen, Stuart G. Gordon, Francine Govers, Niklaus J. Grunwald, Wayne Huang, Kelly L. Ivors, Richard W. Jones, Sophien Kamoun, Konstantinos Krampis, Kurt H. Lamour, Mi-Kyung Lee, W. Hayes McDonald, Monica Medina, Harold J. G. Meijer, Eric K. Nordberg, Donald J. Maclean, Manuel D. Ospina-Giraldo, Paul F. Morris, Vipaporn Phuntumart, Nicholas H. Putnam, Sam Rash, Jocelyn K. C. Rose, Yasuko Sakihama, Asaf A. Salamov, Alon Savidor, Chantel F. Scheuring, Brian M. Smith, Bruno W. S. Sobral, Astrid Terry, Trudy A. Torto-Alalibo, Joe Win, Zhanyou Xu, Hongbin Zhang, Igor V. Grigoriev, Daniel S. Rokhsar, Jeffrey L. Boore
    SCIENCE 313 5791 1261 - 1266 2006年09月 [査読有り][通常論文]
     
    Draft genome sequences have been determined for the soybean pathogen Phytophthora sojae and the sudden oak death pathogen Phytophthora ramorum. Oomycetes such as these Phytophthora species share the kingdom Stramenopila with photosynthetic algae such as diatoms, and the presence of many Phytophthora genes of probable phototroph origin supports a photosynthetic ancestry for the stramenopiles. Comparison of the two species' genomes reveals a rapid expansion and diversification of many protein families associated with plant infection such as hydrolases, ABC transporters, protein toxins, proteinase inhibitors, and, in particular, a superfamily of 700 proteins with similarity to known oomycete avirulence genes.
  • MS Connolly, Y Sakihama, Phuntumart, V, YJ Jiang, F Warren, L Mourant, PF Morris
    CURRENT GENETICS 48 6 356 - 365 2005年12月 [査読有り][通常論文]
     
    A system for the expression of an ATP binding cassette (ABC) transporter from the soybean pathogen Phytophthora sojae is described. Pdr1, an ABC transporter with homology to the pleiotropic drug resistance (PDR) family of transporters, was cloned by primer walking from a P. sojae genomic library. Reverse transcriptase PCR assays showed that the transcript disappeared after encystment of zoospores and was not detected in hyphal germlings in dilute salts, in hyphae growing in liquid V8 media, or in tissue extracts from infected hypocotyls. BLAST analysis of Pdr1 against the P. sojae EST database also revealed that this gene was present only in zoospore libraries. Comparison of the number of hits to Pdr1 with that of a set of housekeeping genes revealed that Pdr1 was expressed at rates two- to threefold higher than other transcripts. To test the hypothesis that Pdr1p functions as a broad substrate membrane transporter, Pdr1 was transformed into yeast mutants deficient in several drug resistance transporters. Yeast mutants transformed with Pdr1 possessed partial drug resistance against only 5 of 17 chemically distinct compounds. Thus, when expressed in yeast, this transporter has a significantly narrower substrate specificity in comparison to the yeast transporters, Pdr5p, Yorlp, and Snq2p.
  • Y Sakihama, T Shimai, M Sakasai, T Ito, Y Fukushi, Y Hashidoko, S Tahara
    ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 432 2 145 - 151 2004年12月 [査読有り][通常論文]
     
    Aphanomyces cochlioides zoospores show chemotaxis to cochliophilin A (5-hydroxy-6,7-methylenedioxyflavone, 1), a host derived attractant, and also respond to 5,7-dihydroxyflavone (2) known as an equivalent chemoattractant. To investigate the chernotactic receptors in the zoospores, we designed photoaffinity probes 4'-azido-5,7-dihydroxyflavone (3) and 4'-azido-7-O-biotinyl-5hydroxyflavone (4) considering chemical structure of 2. Both 3 and 4 had zoospore attractant activity which was competitive with that of 1. When zoospores were treated with the biotinylated photoaffinity probe followed by UV irradiation and streptavidin-gold or peroxidase-conjugated streptavidin, probe-labeled proteins were detected on the cell membrane. This result indicated that the 1-specific-binding proteins, a candidate for hypothetical cochliophilin A receptor, were localized on the cell membrane of the zoospores. This is the first experimental evidence of flavonoid-binding proteins being present in zoospores, using chemically synthesized azidoflavone as photoaflinity-labeling reagent. (C) 2004 Elsevier Inc. All rights reserved.
  • Y Sakihama, R Tamaki, H Shimoji, T Ichiba, Y Fukushi, S Tahara, H Yamasaki
    FEBS LETTERS 553 3 377 - 380 2003年10月 [査読有り][通常論文]
     
    Peroxynitrite (ONOO-), a reactive nitrogen species, is capable of nitrating tyrosine residue of proteins. Here we show in vitro evidence that plant phenolic compounds can also be nitrated by an ONOO--independent mechanism. In the presence of NaNO2, H2O2, and horseradish peroxidase (HRP), monophenolic p-coumaric acid (p-CA, 4-hydroxycinnamic acid) was nitrated to form 4-hydroxy-3-nitrocinnamic acid. The reaction was completely inhibited by KCN, an inhibitor for HRP. The antioxidant ascorbate suppressed p-CA nitration and its suppression time depended strongly on ascorbate concentration. We conclude that nitrogen dioxide radical (NO2), but not ONOO-, produced by a guaiacol peroxidase is the intermediate for phytophenolic nitration. (C) 2003 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
  • Y Sakihama, S Murakami, H Yamasaki
    BIOLOGIA PLANTARUM 46 1 117 - 119 2003年 [査読有り][通常論文]
     
    Nitrite, as well as the nitric oxide (NO) donor S-nitroso-N-acethylpenisilamine (SNAP), was found to increase the aperture of stoma on Vicia faba leaf peels. The results demonstrated here suggest that the nitrite-dependent NO production pathway would be involved in the signal transduction for stomatal movements.
  • Michael F. Cohen, Yasuko Sakihama, Yojiro C. Takagi, Toshio Ichiba, Hideo Yamasaki
    Molecular Plant-Microbe Interactions 15 9 875 - 882 2002年09月01日 [査読有り][通常論文]
     
    The hrmA gene of the N2-fixing cyanobacterium Nostoc punctiforme functions in repressing the formation of transitory motile filaments, termed hormogonia, by plant-associated vegetative filaments. Here, we report that anthocyanins can contribute to induction of hrmA expression. Aqueous extract from fronds of the fern Azolla pinnata, a host of symbiotic Nostoc spp., was found to be a potent inducer of hrmA-luxAB in N. punctiforme strain UCD 328. The hrmA-luxAB inducing activities of A. pinnata, as well as Azolla filiculoides, were positively correlated with levels of frond deoxyanthocyanins. Analyses of the deoxyanthocyanins in frond extracts revealed, in order of predominance, an acetylated glycoside derivative of luteolinidin (m/z 475) and of apigeninidin (m/z 459) and minor amounts of a second luteolinidin derivative. At up to 150 μM, a purified preparation of deoxyanthocyanins only weakly induced hrmA-luxAB on its own, but mixtures with hrmA-luxAB inducers (A. filiculoides extract or the flavonoid naringin) synergistically doubled to tripled their inducing activities. These results suggest that appropriately localized deoxyanthocyanins could function in plant-mediated mechanisms for repressing Nostoc spp. hormogonium formation.
  • Y Sakihama, MF Cohen, SC Grace, H Yamasaki
    TOXICOLOGY 177 1 67 - 80 2002年08月 [査読有り][通常論文]
     
    Plant phenolic compounds such as flavonoids and lignin precursors are important constituents of the human diet. These dietary phytophenolics have been recognized largely as beneficial antioxidants that can scavenge harmful active oxygen species including O(2)(.-), H(2)O(2), (.)OH, and (1)O(2). Here we review our current understanding of the antioxidant and prooxidant actions of phenolics in plant cells. In plant systems, phytophenolics can act as antioxidants by donating electrons to guaiacol-type peroxidases (GuPXs) for the detoxification of H(2)O(2) produced under stress conditions. As a result of such enzymatic as well as non-enzymatic antioxidant reactions, phenoxyl radicals are formed as the primary oxidized products. Until recently, phenoxyl radicals had been difficult to detect by static electron spin resonance (ESR) because they rapidly change to non-radical products. Application of Zn exerts spin-stabilizing effects on phenoxyl radicals that enables us to analyze the formation and decay kinetics of the radicals. The ESR signals of phenoxyl radicals are eliminated by monodehydroascorbate radical (MDA) reductase, suggesting that phenoxyl radicals, like the ascorbate radical, are enzymatically recycled to parent phenolics. Thus, phenolics in plant cells can form an antioxidant system equivalent to that of ascorbate. In contrast to their antioxidant activity, phytophenolics also have the potential to act as prooxidants under certain conditions. For example, flavonoids and dihydroxycinnamic acids can nick DNA via the production of radicals in the presence of Cu and O(2). Phenoxyl radicals can also initiate lipid peroxidation. Recently, Al, Zn, Ca, Mg and Cd have been found to stimulate phenoxyl radical-induced lipid peroxidation. We discuss the mechanism of phenoxyl radical prooxidant activity in terms of lifetime prolongation by spin-stabilizing agents. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
  • Takahashi S, Tamashiro A, Sakihama Y, Yamamoto Y, Kawamitsu Y, Yamasaki H
    BMC Plant Biology 2 2 1471 - 2229 その他の出版社 2002年04月02日 [査読有り][通常論文]
     
    The tropical plant <i>Ficus microcarpa</i> L. f. cv. Golden Leaves (GL) is a high-light sensitive tropical fig tree in which sun-leaves are yellow and shade-leaves are green. We compared the response of photosynthetic activities to strong light between GL and its wild-type (WT, <i>Ficus microcarpa</i> L. f.). Field measurements of maximum photosystem II (PSII) efficiency (Fv/Fm) of intact sun-leaves in GL showed that photo synthetic activity was severely photoinhibited during the daytime (Fv/Fm = 0.46) and subsequently recovered in the evening (Fv/Fm = 0.76). In contrast, WT did not show any substantial changes of Fv/Fm values throughout the day (between 0.82 and 0.78). Light dependency of the CO<sub>2</sub> assimilation rate in detached shade-leaves of GL showed a response similar to that in WT, suggesting no substantial difference in photosynthetic performance between them. Several indicators of photoinhibition, including declines in PSII reaction center protein (D1) content, Fv/Fm value, and O<sub>2</sub> evolution and CO<sub>2</sub> assimilation rates, all indicated that GL is much more susceptible to photoinhibition than WT. Kinetics of PAM chlorophyll a fluorescence revealed
  • Y Sakihama, S Nakamura, H Yamasaki
    PLANT AND CELL PHYSIOLOGY 43 3 290 - 297 2002年03月 [査読有り][通常論文]
     
    Biological activity of nitric oxide (NO) production was investigated in the unicellular green alga Chlamydomonas reinhardtii. An NO specific electrode detected a rapid increase in signal when nitrite (NO(2)(-)) was added into a suspension of C. reinhardtii intact cells in the dark. The addition of KCN or the NO quencher bovine hemoglobin completely abolished the signal, verifying that the nitrite-dependent increase in signal is due to enzymatic NO production. L-arginine, the substrate for NO synthase, did not induce detectable NO production and the NOS inhibitor N(omega)-vitro-L-arginine showed no inhibitory effect on the nitrite-dependent production of NO. Illuminating cells showed a significant suppressive effect on NO production. When the photosynthetic electron transport inhibitor 3(3,4-dichlorophenyl)-1,1-dimethylurea was present in the suspension, C. reinhardtii cells produced NO after the addition of nitrite even under illumination. Kinetic and microscopic observations, using the intracellular fluorescent NO probe 4,5-diaminofluorescein-2 diacetate, both demonstrated that NO was produced within the cells in response to the addition of nitrite. The Chlamydomonas mutant cc-2929, which lacks nitrate reductase (NR) activity, did not display any of the responses observed in the wild-type cells. The results presented here provide direct in vivo evidence to confirm that NR is involved in the nitrite-dependent NO production in the green alga.
  • M. Papina, Y. Sakihama, C. Bena, R. Van Woesik, H. Yamasaki
    Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology 131 4 767 - 774 2002年 [査読有り][通常論文]
     
    This study characterized the spectral properties of Acropora tenuis, A. nasuta, A. secale, and A. aspera, all of which showed strong colorful fluorescence under ultraviolet light-A (black light). The emission maxima of fluorescence from the intact corals were 517, 482, 484, and 514 nm in A. tenuis, A. nasuta, A. secale, and A. aspera, respectively. Using a soluble fraction of cell-free extract of the corals, we applied a method of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to separate each fluorescent protein component contained in the corals. Green fluorescent bands were detected in all Acropora examined, although their apparent molecular mass and relative content were different. A. aspera had two orange bands in addition to the green one. The major excitation and emission peaks of the orange fluorescence bands were almost identical (476 and 478 nm), however, they were discernible by their spectral profiles and molecular masses. Some biochemical properties of the highly fluorescent proteins of Acropora are described and implications of the results are discussed. © 2002 Elsevier Science Inc. All rights reserved.
  • Y Sakihama, H Yamasaki
    BIOLOGIA PLANTARUM 45 2 249 - 254 2002年 [査読有り][通常論文]
     
    Using the whole plant and model systems, we demonstrate that the aluminum ions (Al3+) stimulate phenolic-dependent lipid peroxidation, Lipid peroxidation in barley (Hordeum vulgare L. cv. Donor) roots was 30% higher under AlCl3 treatment than without Al, Major decomposition product of lipid peroxidation was 4-hydroxynonenal (4-HNE) but not thiobarbituric acid reactive substances (TBARS), a widely used markers for lipid peroxidation. Similarly, AlCl3 stimulated lipid peroxidation of soybean liposomes in the presence of chlorogenic acid (CGA) and H2O2/horseradish peroxidase system which can oxidize phenolics. Al3+ was found to enhance lipid peroxidation induced by oxidized CGA. Intermediates of lignin biosynthesis in plants, including p-coumaric acid, ferulic acid, sinapic acid and coniferyl alcohol, also showed similar effects. These results suggest that Al3+ has a potential to induce oxidative stress in plants by stimulating the prooxidant nature of endogenous phenolic compounds.
  • H Yamasaki, H Shimoji, Y Ohshiro, Y Sakihama
    NITRIC OXIDE-BIOLOGY AND CHEMISTRY 5 3 261 - 270 2001年06月 [査読有り][通常論文]
     
    Plant nitrate reductase (NR) produces nitric oxide (NO) when nitrite is provided as the substrate in the presence of NADH [H. Yamasaki and Y. Sakihama (2000) FEBS Lett. 468, 89-92]. Using a NR-dependent NO producing system, we investigated the effects of NO on the energy transduction system in plant mitochondria isolated from mung bean (Vigna radiata). Plant mitochondria are known to possess two respiratory electron transport pathways-the cytochrome and alternative pathways. When the alternative pathway was inhibited by n-propyl gallate, the addition of NR strongly suppressed respiratory O-2 consumption driven by the cytochrome pathway. In contrast, the alternative pathway measured in the presence of antimycin A was not affected by NO. The extent of the steady-state membrane potential (Delta psi) generated by respiratory electron transport rapidly declined in response to NO production. The addition of bovine hemoglobin, a quencher of NO, resulted in the recovery of Delta psi to the uninhibited level. Consistent with its inhibition of Delta psi, NO produced by NR strongly suppressed ATP synthesis in the mitochondria. These results provide substantial evidence to confirm that the plant alternative pathway is resistant to NO and support the idea that the alternative pathway may lower respiration-dependent production of active oxygens under conditions where NO is overproduced, (C) 2001 Academic Press
  • Y Sakihama, J Mano, S Sano, K Asada, H Yamasaki
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 279 3 949 - 954 2000年12月 [査読有り][通常論文]
     
    Monodehydroascorbate (MDA) reductase catalyzes the reduction of MDA, the only organic radical substrate for the enzyme reported so far. Here, we show that cucumber MDA reductase is also capable of reducing phenoxyl radicals which are generated by horseradish peroxidase (HRP) with H2O2. The addition of MDA reductase plus NADH suppressed the HRP/H2O2-dependent oxidation of quercetin, accompanied by the oxidation of NADH, The quenching of the quercetin radical by MDA reductase plus NADH was confirmed by ESR. MDA reductase with NADH also suppressed the HRP/H2O2-dependent oxidation of hydroxycinnamates, including ferulic acid, coniferyl alcohol, and chlorogenic acid. Thus, the phenoxyl radicals of plant phenols can be reduced to their respective parent phenols by MDA reductase via a mechanism similar to the reduction of MDA. (C) 2000 Academic Press.
  • H Yamasaki, Y Sakihama
    FEBS LETTERS 468 1 89 - 92 2000年02月 [査読有り][通常論文]
     
    We examined the ability of plant nitrate reductase (NR) to produce nitric oxide (NO) using in vitro assays. Electrochemical and fluorometric measurements both showed that NO is produced by corn NR in the presence of nitrite and NADH at pH 7. The NO production was inhibited by sodium azide, a known inhibitor for NR. During the reaction, absorbance of 2',7'-dichlorodihydrofluorescein increased markedly. This change was completely suppressed by sodium azide, glutathione or depletion of oxygen. We conclude that plant NR produces both NO and its toxic derivative, peroxynitrite, under aerobic conditions when nitrite is provided as the substrate for NR, (C) 2000 Federation of European Biochemical Societies.
  • H Yamasaki, Y Sakihama, S Takahashi
    TRENDS IN PLANT SCIENCE 4 4 128 - 129 1999年04月 [査読有り][通常論文]
  • H Yamasaki, Y Sakihama, N Ikehara
    PLANT PHYSIOLOGY 115 4 1405 - 1412 1997年12月 [査読有り][通常論文]
     
    Recent studies have revealed that dietary flavonoids are potent radical scavengers, acting in a manner similar to ascorbate and alpha-tocopherol. However, it is still not clear whether flavonoids have a similar antioxidative function in plants. We examined the possibility that flavonoids could function as stress protectants in plant cells by scavenging H2O2. Two major flavonoids, quercetin and kaempferol glycosides, were isolated from leaves of the tropical tree Schefflera arboricola Hayata. Both glycosides and aglycones of isolated flavonols were oxidized by H2O2 in the presence of horseradish peroxidase and/or in a soluble fraction of S. arboricola leaf extract. The rates of oxidation were in the order quercetin > kaempferol > quercetin glycoside >> kaempferol glycoside. Judging from the effects of inhibitors such as KCN, p-chloromercuribenzoate, and 3-amino-1H-1,2,4-triazole, we conclude that guaiacol peroxidase in the soluble fraction catalyzes H2O2-dependent oxidation of flavonols. In the flavonol-guaiacol peroxidase reaction, ascorbate had the potential to regenerate flavonols by reducing the oxidized product. These results provide further evidence that the flavonoid-peroxidase reaction can function as a mechanism for H2O2 scavenging in plants.
  • H Yamasaki, H Uefuji, Y Sakihama
    ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 332 1 183 - 186 1996年08月 [査読有り][通常論文]
     
    Red anthocyanin prepared from petals of Hibiscus rosa-sinensis L. was photobleached in the EDTA-riboflavin system. The rate of bleaching monitored at 565 nm depended on the light intensity and EDTA concentrations. Anaerobic conditions and/or addition of superoxide dismutase prevented the bleaching of anthocyanin, whereas mannitol and catalase did not, A similar bleaching was observed under dark conditions in the xanthine-xanthine oxidase system. The results indicate that anthocyanin is bleached by the nonenzymatic reaction with the superoxide radical and suggest that the pigment can function as an antioxidant. The antioxidative efficiency of cyanidin to superoxide was 10-fold higher than that of cyanidin-3-sophoroside as a Hibiscus anthocyanin. (C) 1996 Academic Press, Inc.
  • 崎浜靖子, 池原規勝, 山崎秀雄
    沖縄生物学会誌 33 43 - 48 1995年03月 [査読有り][通常論文]
     
    Two flavonol gylcosides were isolated from the fresh leaves of the tropical tree <i>Schefflera arboricola</i> Hayata. The major flavonol glycosides of <i>S. arboricola</i> were identified to be quercetin and kaempferol glycosides on the basis of thin layer chromatogram and absorption spectral data. The absorption coefficients of these glycosides were estimated to be 14.7 mM<sup>-1</sup>cm<sup>-1</sup> at A<sub>358</sub> (the quercetin glycoside) and 16.0 mM<sup>-1</sup>cm<sup>-1</sup> at A<sub>346</sub> (the kaempferol glycoside).

MISC

講演・口頭発表等

  • 植物色素の抗酸化機能と環境ストレス保護作用  [招待講演]
    崎浜 靖子
    日本生物工学会北日本支部シンポジウム 2016年03月
  • 硝酸還元酵素によるNO生成  [招待講演]
    崎浜靖子, 崎浜靖子
    日本植物生理学会2008年度年会（札幌） 2008年03月 シンポジウム・ワークショップパネル（指名）
  • 高等植物におけるフラボノイドの生理機能  [招待講演]
    崎浜靖子
    日本農芸化学会北海道支部講演会 2005年08月 口頭発表（招待・特別）

所属学協会

  • 日本農芸化学会北海道支部   日本植物生理学会   日本植物学会   日本NO学会   

共同研究・競争的資金等の研究課題

  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2023年04月 -2027年03月 
    代表者 : 若林 斉, 斉藤 昌之, 松下 真美, 米代 武司, 崎浜 靖子, 渡邊 祐介
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2019年10月 -2023年03月 
    代表者 : 木村 淳夫, 橋床 泰之, 崎浜 靖子, 奥山 正幸, 田上 貴祥
     
    我々は世界で初めてメガロ糖（MS）の生産に成功した。性質を調べると、BCS IIに属す化合物（難水溶性・高膜透過性の薬剤や食品素材など）を可溶化する画期的な機能が発見された。またMSは「BCS II化合物を溶質とする糖質水溶化剤」と捉えることもできた。一方、難溶性ベンジル系アゾ色素もBCS IIに属し、かつ「東南アジア諸国における代表的な環境汚染物質」である点に注目し、MSとアゾ分解酵素を組合せることで、実験室レベルではあるが、色素の可溶化と酵素分解に成功した。以上は初めて生産したMS、すなわち従来型MSの知見である。極最近に従来型MSより高機能な新奇MS（新型MS）を発見した。本申請では、新型MSによるアゾ色素の酵素分解を目的とし、現地試験をタイで実施する。最終的な到達目標はアゾ色素の汚染解消（すなわち環境問題克服への貢献）である。 本年度もタイにおける新型コロナウイルス感染症の流行から渡航が困難となり、現地調査の断念を強いられた。タイでの調査が遅れているが、それを補うべき対策を立てて研究を進行させた。以下に本年度の成果を述べる。１）昨年までに我々が明らかにした有望な植物MS（日タイ共通植物由来のMS）に関し、タイ植物の存在確認を相手国研究協力者が行った。２）本MSの存在下でアゾ色素の酵素分解が進行した。３）自然界においてアゾ色素は土壌（色素汚染土壌）の中に存在するため、アゾ色素と土壌を混合させ「色素汚染土壌」のモデル化を図った。４）構築したモデル汚染土壌を滅菌後に、MS可溶化と酵素処理を実施し、アゾ色素分解の確認を行っている。分解量の向上を目指したい。
  • 植物色素ベタレインの活性酸素・活性窒素からの細胞保護機能に関する研究
    公益財団法人 飯島藤十郎記念食品科学振興財団：平成28年度学術研究助成・個人研究
    研究期間 : 2017年04月 -2018年03月 
    代表者 : 崎浜 靖子
  • アカビート色素ベタレインの活性窒素消去活性とその植物ストレス応答における機能
    公益財団法人 栗林育英学術財団：研究助成（個人研究の部）
    研究期間 : 2015年10月 -2016年03月 
    代表者 : 崎浜 靖子
  • 日本学術振興会：科学研究費助成事業 基盤研究(A)
    研究期間 : 2008年 -2011年 
    代表者 : 橋床 泰之, 原口 昭, 木村 淳夫, 崎浜 靖子, 玉井 裕, 松浦 英幸
     
    インドネシア・カリマンタンの負荷土壌に対処できる根圏微生物コンソーシアムについて検討した。中強酸性泥炭湿地に自生するチガヤ根面から分離したBurkholderia属細菌A-KA株にSphingomonas属など根圏微生物の増殖を促進する活性が認められ,また,酸性硫酸塩土壌耐性イネ根から分離したBurkholderia mimosarumにはイネ苗立枯細菌病を引き起こすBurkholderia plantarii々のトロポロン産生を抑え,種子付着菌を根圏で増殖させる能力を示した。熱帯のポリフェノールリッチな土壌とフタバガキ幼木根圏から分離した細菌群からは,ピロガロール基をもつポリフェノール存在でインドール分解活性が急激に活性化されるBurkholderia属細菌を見いだした。共生系では,ソテツ根と窒素固定性シアノバクテリアNostoc punchiformeとのサンゴ状根共生において,シアノバクテリア誘引に必要な運動性付与がジアシルグリセロールの一分子種で起こることを明らかにした。また,宿主の土壌伝播病原菌感染を抑制する根圏細菌Psuedomonas jessenii EC-S101株やTrichoderma sp. PS1-7株の放出するシグナル物質の化学構造とその機能性を解明した。根圏での脱窒制御に関しては,熱帯泥炭地の亜酸化窒素生成菌では亜酸化窒素還元酵素(N2OR)遺伝子が欠損しており,また,様々な植物二次代謝産物が根圏での脱窒能に影響を与えることが明らかにされた。
  • 日本学術振興会：科学研究費補助金（若手研究B)
    研究期間 : 2006年04月 -2008年03月 
    代表者 : 崎浜 靖子
     
    ダイズ茎疫病菌Phytophthora sojae及びホウレンソウ根腐れ病菌Aphanomyces cochlioidesの遊走子細胞は、宿主植物から滲出する二次代謝産物フラボノイドを認識し、被嚢化、発芽という感染時形態変化を起こす。被嚢化は物理的刺激によっても起こるが、宿主認識物質が存在しない場合には発芽せずに遊走子を再生するため感染には至らない。しかし、動植物の細胞内シグナル物質として知られている一酸化窒素(NO)によって宿主認識物質なしでも遊走子の被嚢化・発芽が促進されることが観察された。また、P. soae遊走子をNOの消去剤であるcarboxy-PTIOで処理すると発芽した菌糸の伸長が有意に阻害された。これらの結果は、菌糸の伸長にNOが関与していることを示している。また、被嚢化後、発芽または再生遊走子のどちらに分化するのかそのギアチェンジに関わる遺伝子を探索するため、発芽を誘引する宿主認識物質処理(A)と遊走子再生を誘引する物理的刺激(B)によって被濃化させた胞子に特異的に発現している遺伝子を調べた。その結果、(A)では細胞骨格維持に関わる膜タンパク質、(B)ではアクチン構築に関与するタンパク質をコードする遺伝子が発現していた。 本研究課題の遂行により、卵菌においてNO依存性シグナル伝達系が宿主感染機構に重要な役割を果たしている可能性が示唆され、宿主感染成立のキーステップである被嚢化から発芽への細胞分化に関わる遺伝子が見いだされた。これらの結果は感染システムのさらなる解明と新たな防除法の確立に応用できる重要な知見であると期待される。
  • 植物病原性卵菌類の感染時シグナル伝達機構について
    科学研究費補助金（若手研究B)
    研究期間 : 2004年04月 -2006年03月 
    代表者 : 崎浜 靖子
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2004年 -2005年 
    代表者 : 崎浜 靖子
     
    植物病原性卵菌類であるAphanomyces及びPhytophthoraは、遊泳能力を持つ遊走子細胞を形成することにより宿主植物に到達し感染する。その際、遊走子は被嚢化しさらに被嚢胞子の発芽が起こる。この遊走子形態変化は宿主感染機構において重要な過程であるがその分子機構については不明な点が多い。本研究では、遊走子の形態変化分子機構解明とその制御に関わる基礎的知見を得ることを目的とし、動植物で様々な生理作用を示す一酸化窒素(NO)の遊走子被嚢化及び発芽に対する影響を調べた。 Aphanomyces cochlioides及びPhytophthora sojae遊走子を自発的NO発生試薬であるNOC5またはNOR3で処理したところ、被嚢化及び発芽率が有意に上昇した。また、哺乳類型NO合成酵素(NOS)の基質であるアルギニンでも遊走子の発芽促進が観察されたため、DOE Joint Genome InstituteのP.sojaeゲノムデータベースを用いNOS様遺伝子の探索を行ったところ、アミノ酸配列で70%の相同性を持つ遺伝子が存在した。しかし、この遺伝子はオキシダーゼドメインを欠いており、同遺伝子を導入した大腸菌ではアルギニン由来のNO生成活性は確認出来なかった。また、脱窒菌や一部の真菌では脱窒過程においてNOを生成することが報告されていることから、脱窒に関わる酵素群遺伝子についても検索したところ、亜硝酸をNOに変換する亜硝酸還元酵素と高い相同性をもつ遺伝子が見つかった。 本研究課題の遂行により、A.cochlioides及びP.sojae遊走子の被嚢化及び発芽機構にNOが大きな影響を与える事が分かった。卵菌類の遊走子は宿主植物に到達できなかった場合は被嚢化しても発芽せずに再度遊走子を作りだすが、NOによって遊走子を強制的に被嚢化・発芽させる"自殺発芽誘導法"等の新たな駆除法の開発への応用が期待される。
  • 日本学術振興会：科学研究費助成事業 基盤研究(A)
    研究期間 : 2002年 -2004年 
    代表者 : 田原 哲士, 吉原 照彦, 橋床 泰之, 福士 幸治, 崎浜 靖子
     
    1)ホウレンソウ根腐れ病菌の宿主特異的遊走子誘引物質cochliophilin Aに対するレセプターを特定するための、光アフィニティー部位とビオチン結合型のプローブを用いた実験で、遊走子の膜画分からプローブとの結合蛋白質として約40および70KDaの電気泳動バンドを検出した.フラボノイドに対する初めての特異的な受容体蛋白質である可能性が十分にある.これらの蛋白質を分離、アミノ酸配列を部分的に決定し、遺伝子を特定する方向性が定まった. 2)各種顕微鏡技術を駆使し、ホウレンソウ根腐れ病菌遊走子の細胞内情報伝達系の解析を試みた.微細形態の経時的変化の観察や細胞内成分の選択的蛍光染色法などを併用して、細胞内で生起する事象の可視化に努めた.その結果、nicotinamideによる被嚢化の初期の段階から遊走子再生への過程と被嚢胞子成熟への分岐が、繊維状アクチンの顕著な構造上の違いを伴うことを発見した.この過程は、根腐れ病菌の生活環展開のキーポイントと目されており、研究展開への重要な手懸かりになると思われる. 3)特異的な情報伝達物質あるいは情報伝達系をターゲットとした卵菌類の生物合理的な制御法の検討を行った.根腐れ病菌の宿主あるいは非宿主植物の根圏から分離した150種のバクテリアのうち約5%が根腐れ病菌の増殖菌糸に形態異常を惹起し、さらに、顕著な形態変化を誘起するものはサトウダイコンの根腐れ病発現を効果的に抑制することを見いだした.この方法論は土壌病原菌の微生物制御資材をスクリーニングする際に極めて有効なものであると提案した。 4)遊走子の挙動、形態変化を指標に生理活性物質の探索と作用様式の解明を試みた.ホウレンソウ根腐れ病菌の宿主特異的遊走子誘引物質cochliophilon Aは、アカザ科ホウレンソウ、サトウダイコン、シロザにのみ存在が知られていた.今回、罹病性のヒユ科ケイトウ(Celosia cristata)の芽生えにcochliophilon Aが存在することを、初めての発見した.ギンナン未熟果皮のアナカルド酸関連化合物やLannea coromandelicaの縮合タンニンが遊走子の遊泳を阻害し、細胞膜を破壊させる作用を見いだし、その作用特性を検討した.
  • 日本学術振興会：科学研究費助成事業 特別研究員奨励費
    研究期間 : 2000年 -2001年 
    代表者 : 崎浜 靖子
  • 日本学術振興会：科学研究費助成事業 特定領域研究(C)
    研究期間 : 2000年 -2000年 
    代表者 : 山崎 秀雄, 崎浜 靖子
     
    生物の種多様性は熱帯・亜熱帯域の低緯度地域で高く、高緯度地域になるに従って減少することが知られている。しかし、依然として種の多様性を生み出すメカニズムは不明である。本研究計画では、沖縄に自生しているクロトンを用いて、多様性創出における転移因子の役割について基礎的知見を得ることを目的とした。6系統のクロトンについて、ヨナグニサン(蛾)Mariner-Like Element(MLE)配列中の転移酵素に対するコンセンサスプライマー用いPCR増幅を行った。その結果、用いた全ての系統において予測される500bpのバンドが検出され、MLEの存在が示唆された。RNA型転移因子の一つであるTy1/Copiaの逆転写酵素の配列に対するdegenerate PCRをおこなった結果、約300bpのPCRクローンが得られ、ワタのTy1/Copiaレトロポゾンと68%の相同性を示した。マングローブを含む樹木性熱帯植物27種の中で、デイゴとフクギからクロトンと同様なMLEのPCRクローンが得られた。今回、同所的に生息する昆虫のMLEが植物であるクロトンに見いだされたことと、クロトン以外の熱帯植物にも同様な配列が認められたことは、MLEが種を越えて同所的に水平伝播していることを強く示唆している。

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