Development of basic technology for metabolic engineering of Arthrospira platensis (Spirulina)

• Project/Area Number: 26870472
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Aquatic life science; Design and evaluation of sustainable and environmental conscious system
• Research Institution: Tokyo Metropolitan University
• Principal Investigator: Ehira Shigeki 首都大学東京, 理工学研究科, 准教授 (90548132)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 微細藻類 / スピルリナ / 形質転換 / 制限修飾系 / バイオ燃料 / メチル化

Outline of Final Research Achievements

In this study, we developed genetic engineering technology indispensable for metabolic engineering of the cyanobacterium Arthrospira platensis (common name: Spirulina) which is one of the microalgae closest to practical use as biomass resources. The biggest barrier in the transformation of Spirulina is that the DNA introduced from the outside of cells is degraded by 11 Type II restriction enzymes and multiple Type I restriction enzymes. In this study, we developed methods for protection of DNA from these restriction enzymes and established the basic technology for the transformation of Spirulina.

Research Products

• [Journal Article] Transcriptional activation of glycogen catabolism and oxidative pentose phosphate pathway by NrrA facilitates cell survival under nitrogen starvation in the cyanobacterium Synechococcus sp. strain PCC 7002. (2018)
  • Author(s): Shimmori Y, Kanesaki Y, Nozawa M, Yoshikawa H, and Ehira S.
  • Journal Title: Plant and Cell Physiology Volume: 印刷中 Issue: 6 Pages: 1225-1233
  • DOI: 10.1093/pcp/pcy059
  • Peer Reviewed
• [Journal Article] The nitrogen-regulated response regulator NrrA is a conserved regulator of glycogen catabolism in β-cyanobacteria (2017)
  • Author(s): Ehira Shigeki、Shimmori Yuka、Watanabe Satoru、Kato Hiroaki、Yoshikawa Hirofumi、Ohmori Masayuki
  • Journal Title: Microbiology Volume: 163 Issue: 11 Pages: 1711-1719
  • DOI: 10.1099/mic.0.000549
  • Peer Reviewed / Open Access
• [Journal Article] Application of CRISPR Interference for Metabolic Engineering of the Heterocyst-Forming Multicellular Cyanobacterium Anabaena sp. PCC 7120 (2017)
  • Author(s): Higo Akiyoshi、Isu Atsuko、Fukaya Yuki、Ehira Shigeki、Hisabori Toru
  • Journal Title: Plant and Cell Physiology Volume: 59 Issue: 1 Pages: 119-127
  • DOI: 10.1093/pcp/pcx166
  • Peer Reviewed
• [Journal Article] Spatial separation of photosynthesis and ethanol production by cell type-specific metabolic engineering of filamentous cyanobacteria (2017)
  • Author(s): Ehira Shigeki、Takeuchi Takuto、Higo Akiyoshi
  • Journal Title: Applied Microbiology and Biotechnology Volume: 102 Issue: 3 Pages: 1523-1531
  • DOI: 10.1007/s00253-017-8620-y
  • 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] 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] Regulation of genes involved in heterocyst differentiation in the cyanobacterium Anabaena sp. strain PCC 7120 by a group 2 sigma factor SigC (2015)
  • Author(s): Ehira, S., Miyazaki, S.
  • Journal Title: Life Volume: 5 Issue: 1 Pages: 587-603
  • DOI: 10.3390/life5010587
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Sucrose synthesis in the nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120 is controlled by the two-component response regulator OrrA (2014)
  • Author(s): Ehira, S., Kimura, S., Miyazaki, S., Ohmori, M.
  • Journal Title: Appl. Environ. Microbiol. Volume: 80 Issue: 18 Pages: 5672-5679
  • DOI: 10.1128/aem.01501-14
  • Peer Reviewed
• [Journal Article] Fatty alcohols can complement functions of heterocyst specific glycolipids in Anabaena sp. PCC 7120. (2014)
  • Author(s): Halimatul, H.S.M., Ehira, S., Awai, K.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 450 Issue: 1 Pages: 178-183
  • DOI: 10.1016/j.bbrc.2014.05.093
  • Peer Reviewed
• [Presentation] シアノバクテリアにおける多様な分化細胞を生み出す分子機構の解明 (2018)
  • Author(s): 肥田真太朗，得平茂樹
  • Organizer: 第12回日本ゲノム微生物学会年会
• [Presentation] 糸状性シアノバクテリアの細胞種特異的代謝工学による嫌気的バイオ燃料生産 (2018)
  • Author(s): 得平茂樹，肥後明佳，竹内卓人
  • Organizer: 第59回日本植物生理学会年会
• [Presentation] Redox regulation of cellular differentiation in cyanobacteria (2018)
  • Author(s): Ehira, S.
  • Organizer: CLS, Tokyo Tech. International Forum 2018
  • Int'l Joint Research / Invited
• [Presentation] igment composition of a cultivated strain of the cyanobacterium Phormidesmis priestleyi isolated from cryoconite on Qaanaaq Glacier, northwest Greenland (2018)
  • Author(s): Kizawa, A., Uetake, J., Kosugi, M., Ehira, S., and Takeuchi, N.
  • Organizer: International Symposium on Cryosphere and Biosphere
  • Int'l Joint Research
• [Presentation] シアノバクテリアAnabaena sp. PCC 7120におけるヘテロシスト分化にともなう窒素固定能の発現制御機構 (2017)
  • Author(s): 小池洋輔、栗尾洋平、兼崎友、吉川博文、得平茂樹
  • Organizer: 第11回日本ゲノム微生物学会年会
  • Place of Presentation: 神奈川
  • Year and Date: 2017-03-02
• [Presentation] ヘテロシストを形成しないシアノバクテリアArthrospira platensis NIES-39におけるhetR遺伝子の機能解析 (2017)
  • Author(s): 小池玲示、得平茂樹
  • Organizer: 第11回日本ゲノム微生物学会年会
  • Place of Presentation: 神奈川
  • Year and Date: 2017-03-02
• [Presentation] シアノバクテリアにおいてグリコーゲン分解が窒素飢餓での生存に必須である (2017)
  • Author(s): 新森友香，兼崎友，吉川博文，得平茂樹
  • Organizer: 第8回日本光合成学会年会
• [Presentation] 酸素発生型光合成で駆動する嫌気発酵プロセス (2017)
  • Author(s): 得平茂樹
  • Organizer: 第8回日本光合成学会年会
• [Presentation] シアノバクテリアにおける窒素飢餓環境への適応機構 (2016)
  • Author(s): 得平茂樹，新森友香
  • Organizer: 日本植物学会第80回大会
  • Place of Presentation: 沖縄
  • Year and Date: 2016-09-16
• [Presentation] シアノバクテリアにおける乾燥耐性獲得機構 (2016)
  • Author(s): 得平茂樹，木村聡，佐藤未歩，大森正之
  • Organizer: 第7回日本光合成学会年会
  • Place of Presentation: 東京
  • Year and Date: 2016-05-27
• [Presentation] 窒素飢餓環境に対するシアノバクテリアの生存戦略の比較解析 (2016)
  • Author(s): 得平茂樹
  • Organizer: 第10回日本ゲノム微生物学会年会
  • Place of Presentation: 東京工業大学（東京）
  • Year and Date: 2016-03-04
• [Presentation] 窒素飢餓環境に対するシアノバクテリアの生存戦略 (2015)
  • Author(s): 得平茂樹
  • Organizer: ラン藻の分子生物学2015
  • Place of Presentation: かずさアカデミアホール（千葉）
  • Year and Date: 2015-11-17
  • Invited
• [Presentation] ヘテロシストパターン形成を制御するセリン・スレオニンキナーゼPknHの機能解析 (2015)
  • Author(s): 片山瑳紀、得平茂樹
  • Organizer: ラン藻の分子生物学2015
  • Place of Presentation: かずさアカデミアホール（千葉）
  • Year and Date: 2015-11-16
• [Presentation] シアノバクテリアにおける分化細胞の機能発現を制御する転写因子DevH (2015)
  • Author(s): 小池洋輔、栗尾洋平、兼崎友、吉川博文、得平茂樹
  • Organizer: ラン藻の分子生物学2015
  • Place of Presentation: かずさアカデミアホール（千葉）
  • Year and Date: 2015-11-16
• [Presentation] 比較機能解析によるシアノバクテリアの環境適応機構の解明 (2015)
  • Author(s): 得平茂樹
  • Organizer: 第30回日本微生物生態学会大会
  • Place of Presentation: 土浦亀城プラザ（茨城）
  • Year and Date: 2015-10-20
• [Presentation] Regulation of compatible solute synthesis in the heterocystous cyanobacterium Anabaena sp. PCC 7120 (2015)
  • Author(s): Ehira, S., Kimura, S., Fan, X.Y., Miyazaki, S. and Ohmori, M.
  • Organizer: 15th International Symposium on Phototrophic Prokaryotes
  • Place of Presentation: Tubingen (Germany)
  • Year and Date: 2015-08-02
  • Int'l Joint Research
• [Presentation] Variety of dependency on DnaA protein for DNA replication among cyanobacterial lineages (2015)
  • Author(s): Ohbayashi, R., Watanabe, S., Ehira, S., Kanesaki, Y., Chibazakura, T. and Yoshikawa, H.
  • Organizer: 15th International Symposium on Phototrophic Prokaryotes
  • Place of Presentation: Tubingen (Germany)
  • Year and Date: 2015-08-02
  • Int'l Joint Research
• [Presentation] Metabolic engineering of differentiated cells of filamentous cyanobacteria for biofuel production (2015)
  • Author(s): Ehira, S.
  • Organizer: The 7th German-Japanese Binational Seminar
  • Place of Presentation: 熱海
  • Year and Date: 2015-03-24
  • Invited
• [Presentation] Anabaena sp. PCC 7120におけるcyAbrBによるgnd遺伝子の発現制御 (2015)
  • Author(s): 西山英里，中村康太，日原由香子，得平茂樹
  • Organizer: 第9回日本ゲノム微生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 2015-03-07
• [Presentation] シアノバクテリアAnabaena sp. PCC 7120のヘテロシスト分化における転写制御因子DevHの機能解析 (2015)
  • Author(s): 栗尾洋平，得平茂樹
  • Organizer: 第9回日本ゲノム微生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 2015-03-06 – 2015-03-07
• [Presentation] Cell-specific metabolic engineering of filamentous cyanobacteria for biofuel production (2015)
  • Author(s): Ehira, S.
  • Organizer: Tokyo Tech-HHU Düsseldorf Joint Symposium on Photosynthesis as a New Chemical Resource
  • Place of Presentation: 東京
  • Year and Date: 2015-03-04
  • Invited
• [Presentation] シアノバクテリアにおける乾燥適応の分子機構 (2014)
  • Author(s): 得平茂樹，Fan Xing-Yan，木村聡，大森正之
  • Organizer: 環境微生物系学会合同大会2014
  • Place of Presentation: 浜松
  • Year and Date: 2014-10-22
• [Presentation] シアノバクテリアにおける細胞特異的な代謝改変によるバイオ燃料生産 (2014)
  • Author(s): 得平茂樹
  • Organizer: 第66回日本生物工学会大会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-10
• [Presentation] シアノバクテリアにおけるグリコーゲン代謝制御の比較解析 (2014)
  • Author(s): 得平茂樹，藤木耕平，西山英里
  • Organizer: 第5回日本光合成学会年会
  • Place of Presentation: 奈良
  • Year and Date: 2014-05-30 – 2014-05-31
• [Presentation] ヘテロシスト形成に伴う遺伝子発現とチラコイド膜光化学系変化の同時タイムラプス顕微分光 (2014)
  • Author(s): 野末秀穂，佐藤直樹，豊島正和，得平茂樹，寺嶋正秀，熊崎茂一
  • Organizer: 第5回日本光合成学会年会
  • Place of Presentation: 奈良
  • Year and Date: 2014-05-30 – 2014-05-31