Molelcular mechanisms of exopolysaccharide production in edible cyanobacterium, Aphanothece sacrum (Suizenji-Nori)

• Project/Area Number: 15K07557
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Aquatic bioproduction science
• Research Institution: Fukui Prefectural University
• Principal Investigator: OHKI Kaori 福井県立大学, 海洋生物資源学部, 名誉教授 (90101104)
• Co-Investigator(Kenkyū-buntansha): 兼崎 友 静岡大学, グリーン科学技術研究所, 特任助教 (70380293)
• Co-Investigator(Renkei-kenkyūsha): YOSHIKAWA Sinya 福井県立大学, 海洋生物資源学部, 准教授 (20405070)
• Research Collaborator: KANEKO Tatsuo 北陸先端技術大学院大学, 環境エネルギー領域, 教授 ; TAKAGI Keiichi 若狭湾エネルギー研究センター, 生物資源研究室, 室長
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: スイゼンジノリ / Aphanothece sacrum / 単細胞ラン藻 / 細胞外多糖合成 / 細胞外多糖合成関連遺伝子 / 全ゲノム解析 / exopolysaccharide (EPS) / シアノバクテリア / 細胞外多糖 / Exopolysaccharides / 細胞外多糖合成遺伝子 / Exopolysaccharaides / 細胞外多糖生産 / 細胞外多糖生産遺伝子 / Aphanothece sucrum

Outline of Final Research Achievements

Unicellular cyanobacterium Aphanothece sacrum (Suizenji-Nori) has been cultured in open-air aqua-farms for more than 300 years as a food source in Japan. At present, the exopolysaccharide (EPS) of A. sacrum attracts as a useful bio-resource. To elucidate the molecular mechanisms related to EPS synthesis and export in A. sacrum, two clonal strains were isolated from a traditional aqua-farm and used. Results revealed that (1) the optimum conditions for growth and EPS synthesis were at around 20C and the light intensity of ca. 80 μmole/sq.m・sec (12h light/12h dark cycles) , (2)purified EPS consisted of five major monosaccharids with a large molecular weight of ca. 10,000 kDa. (3)creation of mutants related to EPS synthesis was tried using heavy ion carbon beam irradiation. (4) the whole genome sequence of the two strains were determined. Analysis of putative genes involved in EPS synthesis and export suggests that the Wzy-dependent pathway is participate in EPS production of A. sacrum.

Research Products

• [Journal Article] Draft Genome Sequence of the Nitrogen-Fixing and Hormogonia-Inducing Cyanobacterium Nostoc cycadae Strain WK-1, Isolated from the Coralloid Roots of Cycas revoluta. (2018)
  • Author(s): Kanesaki Y, Hirose M, Hirose Y, Fujisawa T, Nakamura Y, Watanabe S, Matsunaga S, Uchida H, and Murakami A.
  • Journal Title: Genome Announcements Volume: 6 Issue: 7
  • DOI: 10.1128/genomea.00021-18
  • NAID: 120006488529
  • Peer Reviewed / Open Access
• [Journal Article] Complete Genome Sequence of the Nonheterocystous Cyanobacterium Pseudanabaena sp. ABRG5-3 (2018)
  • Author(s): Tajima Naoyuki、Kanesaki Yu、Sato Shusei、Yoshikawa Hirofumi、Maruyama Fumito、Kurokawa Ken、Ohta Hiroyuki、Nishizawa Tomoyasu、Asayama Munehiko、Sato Naoki
  • Journal Title: Genome Announcements Volume: 6 Issue: 6
  • DOI: 10.1128/genomea.01608-17
  • NAID: 120006457123
  • Peer Reviewed / Open Access
• [Journal Article] ParA-like protein influences the distribution of multi-copy chromosomes in cyanobacterium Synechococcus elongatus PCC 7942. (2018)
  • Author(s): Watanabe S, Noda A, Ohbayashi R, Uchioke K, Kurihara A, Nakatake S, Morioka S, Kanesaki Y, Chibazakura T, Yoshikawa H
  • Journal Title: Microbiology Volume: 164 Issue: 1 Pages: 45-56
  • DOI: 10.1099/mic.0.000577
  • Peer Reviewed / Open Access
• [Journal Article] Physiological properties and genetic analysis related to exopolysaccharide (EPS) production in the fresh-water unicellular cyanobacterium Aphanothece sacrum (Suizenji Nori) (2018)
  • Author(s): Ohki1 Kaori, Kanesaki Yu, Suzuki1 Noriaki, Okajima Maiko, Kaneko Tatsuo, Yoshikawa Sinya
  • Journal Title: Jornal General Applied Microbiology Volume: 64
  • NAID: 130007608428
  • Peer Reviewed
• [Journal Article] Complete Genome Sequence of a Coastal Cyanobacterium,Synechococcussp. Strain NIES-970 (2017)
  • Author(s): Shimura Yohei、Hirose Yuu、Misawa Naomi、Wakazuki Sachiko、Fujisawa Takatomo、Nakamura Yasukazu、Kanesaki Yu、Yamaguchi Haruyo、Kawachi Masanobu
  • Journal Title: Genome Announcements Volume: 5 Issue: 14
  • DOI: 10.1128/genomea.00139-17
  • Peer Reviewed / Open Access
• [Journal Article] Conserved two-component Hik34-Rre1 module directly activates heat-stress inducible transcription of major chaperone and other genes in Synechococcus elongatus PCC 7942. (2017)
  • Author(s): Kobayashi I., Watanabe S., Kanesaki Y., Shimada T., Yoshikawa H. and Tanaka K.
  • Journal Title: Molecular Microbiology Volume: 104 Pages: 260-277
  • Peer Reviewed
• [Journal Article] CyanoBase: a large-scale update on its 20th anniversary. (2017)
  • Author(s): Fujisawa, T., Narikawa, R., Maeda, S., Watanabe, S., Kanesaki, Y., Kobayashi, K., Nomata, J., Hanaoka, M., Watanabe, M., Ehira, S., Suzuki, E., Awai, K. and Nakamura, Y.
  • Journal Title: Nucleic Acids Research Volume: 45 Issue: D1 Pages: 551-554
  • DOI: 10.1093/nar/gkw1131
  • Peer Reviewed / Open Access
• [Journal Article] Variety of DNA Replication Activity Among Cyanobacteria Correlates with Distinct Respiration Activity in the Dark (2017)
  • Author(s): Ryudo Ohbayashi, Jun-ya Yamamoto, Satoru Watanabe, Yu Kanesaki, Taku Chibazakura, Shin-ya Miyagishima, Hirofumi Yoshikawa
  • Journal Title: Plant and Cell Physiology Volume: 58 (2) Pages: 279-286
  • DOI: 10.1093/pcp/pcw186
  • Peer Reviewed
• [Journal Article] Heavy metal biosorption from aqueous solutions by algae inhabiting rice paddies in Vietnam. (2016)
  • Author(s): Tran HT, Vu ND, Matsukawa M, Okajima M, Kaneko T, Ohki K, Yoshikawa S
  • Journal Title: J Environ Chem Engineer Volume: 4 Pages: 2529-2535
  • Peer Reviewed
• [Journal Article] Heavy metal biosorption from aqueous solutions by algae inhabiting rice paddies in Vietnam (2016)
  • Author(s): Hang,T.T., Ngo,D.V., Matsukawa,M., Okajim,M., Kaneko,T., Ohki,K., Yoshikawa,S.
  • Journal Title: Environ Chem Eng Volume: 4 Pages: 2529-2536
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Ultrastructural analysis of siliceous cell wall regeneration in Triparma Laevis NIES-2565 (Parmales, Stramenopiles) (2016)
  • Author(s): Yamada,K., Yoshikawa,Y., Ohki,K., Ichinomiya,M., Kuwata,A., Motomura,T., Nagasato,C.
  • Journal Title: Phycologia Volume: 55
• [Journal Article] Complete genome sequencing of cyanobacterium Leptolyngbya sp. NIES-3755 Journal of Biotechnology. (2016)
  • Author(s): Hirose Y., Fujisawa T., Ohtsubo Y., Katayama M., Misawa N., Wakazuki S., Shimura Y., Nakamura Y., Kawachi M., Yoshikawa H., Eki T., and Kanesaki Y.
  • Journal Title: Genome Announcements Volume: 4 Issue: 2
  • DOI: 10.1128/genomea.00090-16
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Complete genome sequence of cyanobacterium Fischerella sp. NIES-3754, providing thermoresistant optogenetic tools. (2016)
  • Author(s): Hirose Y., Fujisawa T., Ohtsubo Y., Katayama M., Misawa N., Wakazuki S., Shimura Y., Nakamura Y., Kawachi M., Yoshikawa H., Eki T., and Kanesaki Y.
  • Journal Title: Journal of Biotechnology Volume: 220 Pages: 45-46
  • DOI: 10.1016/j.jbiotec.2016.01.011
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Complete genome sequence of cyanobacterium Nostoc sp. NIES-3756, a potentially useful strain for phytochrome-based bioengineering. (2016)
  • Author(s): Hirose Y., Fujisawa T., Ohtsubo Y., Katayama M., Misawa N., Wakazuki S., Shimura Y., Nakamura Y., Kawachi M., Yoshikawa H., Eki T., and Kanesaki Y.
  • Journal Title: Journal of Biotechnology Volume: 220 Pages: 51-52
  • DOI: 10.1016/j.jbiotec.2015.12.002
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Diversification of DnaA dependency for DNA replication in cyanobacterial evolution. (2016)
  • Author(s): Ohbayashi R., Watanabe S., Ehira S., Kanesaki Y., Chibazakura T., and Yoshikawa H.
  • Journal Title: ISME Journal Volume: 10 Issue: 5 Pages: 1113-1121
  • DOI: 10.1038/ismej.2015.194
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Defense mechanisms of sargassacean species against the epiphytic red alga Neosiphonia harveyi (2015)
  • Author(s): Nakajima,N., Ohki,K., Kamiya,M.
  • Journal Title: J. Phycol. Volume: 51 Pages: 695-705
  • Peer Reviewed
• [Journal Article] Intensive DNA replication and metabolism during the lag phase in cyanobacteria (2015)
  • Author(s): Watanabe S., Ohbayashi R., Kanesaki Y., Saito N., Chibazakura T., Soga T., and Yoshikawa H.
  • Journal Title: PLoS One Volume: 10 Issue: 9 Pages: e0136800-e0136800
  • DOI: 10.1371/journal.pone.0136800
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of spontaneous suppressor mutants from the photomixotrophically-grown pmgA-disrupted mutant in the cyanobacterium Synechocystis sp. PCC 6803 (2015)
  • Author(s): Nishijima, Y., Kanesaki, Y., Yoshikawa, H., Ogawa, T., Sonoike, K., Nishiyama, Y. and Hihara, Y.
  • Journal Title: Photosynth. Res. Volume: - Issue: 2-3 Pages: 465-475
  • DOI: 10.1007/s11120-015-0143-8
  • Peer Reviewed
• [Presentation] シアノバクテリアにおける新規高温耐性株の取得とリシーケンス解析 (2017)
  • Author(s): 兼崎友、樋山智恵美、小澤啓吾、宮澤和己、渡辺智、吉川博文
  • Organizer: 日本農芸化学会 2017年度大会
  • Place of Presentation: 京都府京都市
  • Year and Date: 2017-03-17
• [Presentation] アオコ原因シアノバクテリアPlanktothrix agardhii NIES-204株の完全ゲノム解読 (2017)
  • Author(s): 志村遥平、藤澤貴智、広瀬侑、兼崎友、河地正伸
  • Organizer: 第11回日本ゲノム微生物学会
  • Place of Presentation: 神奈川県藤沢市
  • Year and Date: 2017-03-02
• [Presentation] 長期高温培養によるシアノバクテリア新規高温適応株の取得 (2017)
  • Author(s): 兼崎友, 桶山智恵美、宮澤和己、渡辺智
  • Organizer: NGS現場の会
• [Presentation] シアノバクテリアにおける高温適応進化実験の試み (2017)
  • Author(s): 兼崎友, 桶山智恵美、宮澤和己、渡辺智、吉川博文
  • Organizer: 環境微生物系学会合同大会
• [Presentation] スイゼンジノリ分離株を用いた室内培養と細胞外多糖生産 (2016)
  • Author(s): 大城香, 鈴木謙之, 兼崎友, 岡島麻衣子、金子達男、吉川伸哉
  • Organizer: 日本水産学会中部支部会
  • Place of Presentation: 福井県小浜市
  • Year and Date: 2016-12-03
• [Presentation] スイゼンジノリ(Aphanothece sacrum)の細胞外多糖形成と関連遺伝子の解析 (2016)
  • Author(s): 大城香、鈴木謙之、兼崎友、吉川伸哉
  • Organizer: 日本植物学会第80回大会
  • Place of Presentation: 沖縄県宜野湾市
  • Year and Date: 2016-09-16
• [Presentation] スイゼンジノリ(Aphanothece sacrum)分離株の増殖特性と細胞外多糖生産能の解析 (2016)
  • Author(s): 鈴木謙之・一宮睦雄・岡島麻衣子・金子達雄・吉川伸哉・大城香
  • Organizer: 日本藻類学会
  • Place of Presentation: 日本医科歯科大学、東京
  • Year and Date: 2016-03-18
• [Presentation] 単細胞ラン藻スイゼンジノリの分離株を用いた成長特性と多糖生産の検討 (2015)
  • Author(s): 大城 香・鈴木謙之・牧しづか・吉川伸哉、 岡島麻衣子・金子達男
  • Organizer: 日本植物学会
  • Place of Presentation: 朱鷺メッセ、新潟
  • Year and Date: 2015-09-06
• [Presentation] Growth and exopolysaccharide synthesis of edible freshwater cyanobacterium, _Aphanothece_ _sacrum_ (Suizenji Nori). (2015)
  • Author(s): Ohki,K. Suzuki,N., Maki,S., Yoshikawa,S., Okajima,M., Kaneko,T.
  • Organizer: International Symposium on Photosynthetic Prokaryotes
  • Place of Presentation: Tubingen, Germany
  • Year and Date: 2015-08-02
  • Int'l Joint Research
• [Presentation] Identification of the associated genes for substrain-specific phenotypes of a cyanobacterium, Synechococcus elongatus PCC 7942. (2015)
  • Author(s): Kanesaki Y., Ohbayashi R., Watanabe S., Yoshikawa H.
  • Organizer: International Symposium on Photosynthetic Prokaryotes
  • Place of Presentation: Tubingen, Germany
  • Year and Date: 2015-08-02
  • Int'l Joint Research
• [Patent(Industrial Property Rights)] スイゼンジノリのクローン単藻株を用いた静置培養法による室内閉鎖養殖系 (2015)
  • Inventor(s): 大城香、吉川伸哉
  • Industrial Property Rights Holder: 公立大学法人福井県立大学
  • Industrial Property Rights Type: 特許
  • Patent Publication Number: 2017-035051
  • Filing Date: 2015-08-12
  • Acquisition Date: 2017-02-16
• [Patent(Industrial Property Rights)] スイゼンジノリのクローン単藻株を用いた静置培養法による室内閉鎖養殖系 (2015)
  • Inventor(s): 大城香、吉川伸也
  • Industrial Property Rights Holder: 大城香、吉川伸也
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-08-12