Creation of novel poly-extremozymes by modifying protein surfaces

• Project/Area Number: 25410172
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: NAKAMURA SATOSHI 東京工業大学, 生命理工学研究科, 教授 (50227899)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 極限環境微生物 / 極限酵素 / 分子表面電荷 / タンパク質工学 / 進化分子工学 / 極限環境耐性 / 比活性 / キシラナーゼ / 耐アルカリ性 / 耐塩性 / キチナーゼ / 耐熱性 / 有機溶媒耐性 / アルカリキシラナーゼ

Outline of Final Research Achievements

Alkalitolerancy, thermotolerancy and halotolerancy of a GH family 11 alkaline xylanase, XynJ, were improved by introducing basic amino acids on its protein surface. Halotolerancy and organic solvent-tolerancy of a GH family 18 halotolerant chitinase, ChiN1, were improved by introducing acidic amino acids on its protein surface. Alkalitolerancy and specific activity of a GH family 10 hyper-thermotolerant xylanase, XynTB, were improved by directed evolution.

Research Products

• [Int'l Joint Research] Tsinghua University(China)
• [Journal Article] Cloning of a Chitinase Gene from Newly Isolated Alkaliphilic Nocardiopsis sp. Strain F96 and Extracellular Production of the Recombinant Protein in Escherichia coli (2015)
  • Author(s): Kimiko Endo, Kohei Nagao, Kinuka Toyama, Shunsei Ishida, Tetsuya Fukazawa, Rie Yatsunami, Toshiaki Fukui, Xinhui Xing and Satoshi Nakamura
  • Journal Title: Journal of Japanese Society for Extremophiles Volume: 14 Pages: 21-29
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Complete Biosynthetic Pathway of the C50 Carotenoid Bacterioruberin from Lycopene in the Extremely Halophilic Archaeon Haloarcula japonica (2015)
  • Author(s): Yang Y, Yatsunami R, Ando A, Miyoko N, Fukui T, Takaichi S, Nakamura S
  • Journal Title: J Bacterol Volume: 197 Issue: 9 Pages: 1614-1623
  • DOI: 10.1128/jb.02523-14
  • Peer Reviewed
• [Journal Article] Identification of carotenoids from the extremely halophilic archaeon Haloarcula japonica (2014)
  • Author(s): Rie Yatsunami, Ai Ando, Ying Yang, Shinichi Takaichi, Masahiro Kohno, Yuriko Matsumura, Hiroshi Ikeda, Toshiaki Fukui, Kaoru Nakasone, Nobuyuki Fujita, Mitsuo Sekine, Tomonori Takashina and Satoshi Nakamura
  • Journal Title: Frontiers Microbiology Volume: 5
  • Peer Reviewed / Open Access
• [Journal Article] Enhancement of glycerol utilization ability of Ralstonia eutropha H16 for production of polyhydroxyalkanoates (2014)
  • Author(s): Toshuaki Fukui, Masaharu Mukoyama, Izumi Orita and Satoshi Nakamura
  • Journal Title: Applied Microbiology and Biotechnology Volume: 98 Pages: 7559-7568
  • Peer Reviewed / Open Access
• [Journal Article] Gene Analysis, Expression, and Characterization of an Intracellular &#945;-Amylase from the Extremely Halophilic Archaeon <i>Haloarcula japonica</i> (2013)
  • Author(s): M. Onodera, R. Yatsunami, W. Tsukimura, T. Fukui, K. Nakasone, T. Takashina and S. Nakamura
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 77 Issue: 2 Pages: 281-288
  • DOI: 10.1271/bbb.120693
  • NAID: 10031164729
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Journal Article] Gene expression and characterization of aerotaxis transducer HemAT from extremely halophilicarchaeonHaloarcula japonica TR-1 (2013)
  • Author(s): TeppeiTadikara, ToshitakaMatsubara, Yoshihiro Kubota, Takayuki Kosaka, Takatoshi Ozawa, WataruTsukimura, RieYatsunami, Toshiaki Fukui, Kaoru Nakasone, Tomonori Takashina and Satoshi Nakamura
  • Journal Title: Journal of Japanese Society for Extremophiles Volume: 12 Pages: 29-32
  • Peer Reviewed
• [Presentation] Tactic responses of extremely halophilic archaeon Haloarcula japonica overexpressing its aerotaxis transducers (2015)
  • Author(s): Toshitaka Matsubara, Teppei Tadikara, Rie Yatsunami, Toshiaki Fukui and Satoshi Nakamura
  • Organizer: PACIFICHEM2015
  • Place of Presentation: Honolulu, Hawaii, USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research
• [Presentation] 高度好塩性古細菌由来GHファミリー18キチナーゼのタンパク質工学検討：分子表面上の酸性アミノ酸およびリシンの役割 (2015)
  • Author(s): 遠山絹華, 坂上耕一, 張 楊, 安 然, 八波利恵, 高品知典, 福居俊昭, 中村 聡
  • Organizer: 第29回日本キチン・キトサン学会大会
  • Place of Presentation: 東海大学熊本キャンパス
  • Year and Date: 2015-08-20
• [Presentation] 分子表面に過剰な電荷を導入したアルカリキシラナーゼの種々の塩溶液中での挙動 (2014)
  • Author(s): 長尾浩平，三谷俊介，梅本博仁，八波利恵，福居俊昭，中村 聡
  • Organizer: 第15回極限環境生物学会年会
  • Place of Presentation: 沖縄県今帰仁村
  • Year and Date: 2014-11-01 – 2014-11-03
• [Presentation] 高度好塩性古細菌Halobacterium salinarum NRC-1株由来 GH ファミリー 18キチナーゼの性質検討：分子表面上の酸性アミノ酸およびリシンの役割 (2014)
  • Author(s): 遠山絹華，坂上耕一，張楊，安然，八波利恵，高品知典，福居俊昭，中村 聡
  • Organizer: 第15回極限環境生物学会年会
  • Place of Presentation: 沖縄県今帰仁村
  • Year and Date: 2014-11-01 – 2014-11-03
• [Presentation] Characterization of a haloarchaealchitinase: Effect of aspartates, glutamates and lysines on its protein surface (2013)
  • Author(s): Kinuka Toyama, Koichi Sakagami, Zhang Yang, Ann Ran, Motoaki Sato, KeitaOrishimo, YoshinoriHatori, Rie Yatsunami, TomonoriTakashina, Toshiaki Fukui, Satoshi Nakamura
  • Organizer: 10th Asia-Pacific Chitin & Chitosan Symposium (第10回アジア・太平洋キチンキトサンシンポジュウム)・第27回キチン・キトサンシンポジウム
  • Place of Presentation: 米子コンベンションセンター（鳥取県米子市）
• [Book] 極限環境生命 (2014)
  • Author(s): 伊藤政博，道久則之，鳴海一成，東端啓貴，為我井秀行，國枝武和，伊藤 隆，佐藤孝子，中村 聡
  • Total Pages: 232
  • Publisher: コロナ社
• [Book] 進化分子工学：高速分子進化によるタンパク質・核酸の開発（伏見 譲・監修） (2013)
  • Author(s): 中村 聡
  • Publisher: エヌ・ティー・エス
• [Remarks] 東京工業大学 生命理工学院 中村（聡）・八波研究室ホームページ
  • URL: http://www.nakamura.bio.titech.ac.jp/
• [Remarks] 東京工業大学 大学院生命理工学研究科 生物プロセス専攻 中村聡研究室ホームページ
  • URL: http://www.nakamura.bio.titech.ac.jp/
• [Remarks] 東京工業大学 大学院生命理工学研究科 生物プロセス専攻 中村聡研究室ホームページ
  • URL: http://www.nakamura.bio.titech.ac.jp/