Modification of protein synthesis machinery of silkworms for the production of silk proteins bearing unnatural amino acids

• Project/Area Number: 24688008
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Applied entomology
• Research Institution: National Institute of Agrobiological Sciences
• Principal Investigator: TERAMOTO Hidetoshi 独立行政法人農業生物資源研究所, 新機能素材研究開発ユニット, 主任研究員 (60391562)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥13,780,000 (Direct Cost: ¥10,600,000、Indirect Cost: ¥3,180,000); Fiscal Year 2014: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2013: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2012: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
• Keywords: 昆虫 / 生体機能利用 / バイオテクノロジー / 非天然アミノ酸 / other genkou

Outline of Final Research Achievements

This study aimed at developing a method to incorporate unnatural amino acids bearing functional groups available for selective chemical reactions (click reactions) into silk proteins in order to provide high-performance silk-based materials to biomedical fields. We have succeeded in incorporating unnatural amino acids bearing azide groups (AzPhe) available for click reactions into silk proteins using genetic modification techniques of silkworms. We verified that azide groups incorporated into silk proteins can be selectively modified with functional molecules using click reactions.

Research Products

• [Journal Article] Characterization of <i>Bombyx mori</i> Silk Fiber Incorporating an Unnatural Amino Acid (4-Chlorophenylalanine) (2015)
  • Author(s): 寺本英敏, 中島健一, 小島 桂
  • Journal Title: The Journal of Silk Science and Technology of Japan Volume: 23 Issue: 0 Pages: 27-35
  • DOI: 10.11417/silk.23.27
  • NAID: 130005066407
  • ISSN: 1880-8204, 1881-1698
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Incorporation of methionine analogues into Bombyx mori silk fibroin for click modifications (2015)
  • Author(s): Teramoto H. and Kojima K.
  • Journal Title: Macromolecular Bioscience Volume: in press Issue: 5 Pages: 719-727
  • DOI: 10.1002/mabi.201400482
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] duction of Bombyx mori silk fibroin incorporated with unnatural amino acids (2014)
  • Author(s): Teramoto H. and Kojima K.
  • Journal Title: Biomacromolecules Volume: 15(7) Issue: 7 Pages: 2682-2690
  • DOI: 10.1021/bm5005349
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Residue-specific incorporation of phenylalanine analogues into protein biosynthesis in silkworm cultured cells (2013)
  • Author(s): Hidetoshi Teramoto and Katsura Kojima
  • Journal Title: Journal of Insect Biotechnology and Sericology Volume: 82
  • NAID: 130004935607
  • Peer Reviewed
• [Presentation] Azide-bearing silk as a clickable biopolymer produced by in vivo incorporation of unnatural amino acids (2014)
  • Author(s): Teramoto, H. and Kojima, K.
  • Organizer: The 10th SPSJ International Polymer Conference (IPC 2014)
  • Place of Presentation: つくば国際会議場（茨城県・つくば市）
  • Year and Date: 2014-12-02 – 2014-12-05
• [Presentation] Mutagenesis of Bombyx mori silk fiber by in vivo incorporation of unnatural amino acids into silk fibroin (2014)
  • Author(s): Teramoto, H., Kojima, K., and Nakajima, K.
  • Organizer: International Symposium on Fiber Science and Technology 2014
  • Place of Presentation: ビッグサイト東京ファッションタウン（TFT）ホール（東京都・江東区）
  • Year and Date: 2014-09-28 – 2014-10-01
• [Presentation] 天然にはない官能基を導入したシルク材料の創製 (2014)
  • Author(s): 寺本英敏, 小島 桂, 中島健一
  • Organizer: 平成26年度繊維学会年次大会
  • Place of Presentation: タワーホール船堀（東京都・江戸川区）
  • Year and Date: 2014-06-11 – 2014-06-13
• [Presentation] 非天然アミノ酸を導入したシルクの物性解析 (2014)
  • Author(s): 寺本英敏, 小島 桂, 中島健一
  • Organizer: 第61回日本シルク学会研究発表会
  • Place of Presentation: 蚕糸科学研究所（東京都・新宿区）
  • Year and Date: 2014-05-16
• [Presentation] カイコタンパク質合成系へのフェニルアラニン類縁体の導入 (2013)
  • Author(s): 寺本英敏・小島桂
  • Organizer: 平成25年度蚕糸・昆虫機能利用学術講演会
  • Place of Presentation: 農林水産技術会議事務局(茨城県つくば市)
  • Year and Date: 2013-03-19
• [Presentation] カイコにおけるタンパク質構成アミノ酸レパートリーの拡張 (2013)
  • Author(s): 寺本英敏・小島 桂
  • Organizer: つくば医工連携フォーラム2013
  • Place of Presentation: 産業技術総合研究所(茨城県つくば市)
  • Year and Date: 2013-01-29
• [Presentation] シルクに非天然アミノ酸を導入できる遺伝子組換えカイコの作出
  • Author(s): 寺本英敏, 小島桂, 中島健一
  • Organizer: 平成26年度蚕糸・昆虫機能利用学術講演会-日本蚕糸学会第84回大会-
  • Place of Presentation: 日本大学生物資源学部
• [Presentation] クリック反応による誘導化が可能なシルク材料の創製
  • Author(s): 寺本英敏, 小島桂
  • Organizer: つくば医工連携フォーラム2014
  • Place of Presentation: 産業技術総合研究所（つくば）
• [Presentation] 遺伝子組換えカイコによる非天然アミノ酸含有シルクタンパク質の作出
  • Author(s): 寺本英敏, 小島桂
  • Organizer: 第62回高分子討論会
  • Place of Presentation: 金沢大学
• [Patent(Industrial Property Rights)] トランスジェニックカイコを用いた非天然アミノ酸含有タンパク質の製造方法 (2018)
  • Inventor(s): 寺本 英敏, 小島 桂, 飯塚 哲也
  • Industrial Property Rights Holder: (国研)農研機構
  • Industrial Property Rights Type: 特許
  • Acquisition Date: 2018
• [Patent(Industrial Property Rights)] トランスジェニックカイコ、および該カイコを用いた非天然アミノ酸含有タンパク質の製造方法 (2013)
  • Inventor(s): 寺本英敏，小島 桂，飯塚哲也
  • Industrial Property Rights Holder: 寺本英敏，小島 桂，飯塚哲也
  • Industrial Property Rights Type: 特許
  • Filing Date: 2013-07-11