Development of glycosyl modification machinery by sequential multiple infections of baculoviruses

• Project/Area Number: 21510220
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Living organism molecular science
• Research Institution: Akita University
• Principal Investigator: GOTOH Takeshi 秋田大学, 大学院・工学資源学研究科, 教授 (10215494)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2010: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2009: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 生物活性分子の設計 / 合成 / 糖鎖化学 / バイオテクノロジー / 昆虫細胞 / バキュロウイルス / 糖転移酵素 / 糖鎖修飾

Research Abstract

Several recombinant baculoviruses that contained c DNAs of glycosyltransferase soluble domains under the control of gp64 early promoter were constructed, in order to develop a glycosyl modification machinery in insect cells, in which multiple glycosyltransferases were sequentially expressed. In addition, a novel method was proposed that the proteolytic degradation of recombinant proteins expressed in baculovirus-infected insect cells was efficiently inhibited.

Research Products

• [Journal Article] Purification and characterization of aspartic protease derived from Sf9 insect cells (2010)
  • Author(s): Gotoh, T., Ono, H., Kikuchi, K.-I., Nirasawa, S. Takahashi, S
  • Journal Title: Biosci. Biotechnol. Biochem Volume: 74(10) Pages: 2154-2157
• [Journal Article] Active human renin production using a baculovirus expression vector system : An effective method for preventing excessive proteolytic degradation of recombinant proteins (2010)
  • Author(s): Gotoh, T., Kikuchi, K.-I., Takahashi, S
  • Journal Title: J. Chem. Eng. Jpn Volume: 43(7) Pages: 603-607
• [Journal Article] Prorenin processing enzyme (PPE) produced by baculovirus-infected Sf-9 insect cells : PPE is the cysteine protease encoded in the AcMNPV gene (2010)
  • Author(s): Gotoh, T., Awa, H., Kikuchi, K.-I., Nirasawa, S., Takahashi, S
  • Journal Title: Biosci. Biotechnol. Biochem Volume: 74(2) Pages: 370-374
• [Journal Article] Active human renin production using a baculovirus expression vector system : An effective method for preventing excessive proteolytic degradation of recombinant proteins (2010)
  • Author(s): Takeshi Gotoh, 他2名
  • Journal Title: J.Chem.Eng.Jpn Volume: 43 Pages: 603-607
  • Peer Reviewed
• [Journal Article] Expression and in situ processing of human prorenin to active renin in baculovirus-infected Sf-9 insect cell cultures under several infective conditions (2009)
  • Author(s): Gotoh, T., Awa, H., Kikuchi, K.-I., Takahashi, S
  • Journal Title: Biochem. Eng. J Volume: 43(2) Pages: 216-220
• [Presentation] バキュロウイルス感染Sf9昆虫細胞によるヒトインターロイキン-2の生産 (2011)
  • Author(s): 進藤祐宜, 後藤猛, 菊地賢一
  • Organizer: 秋田応用生命科学研究会講演会
  • Place of Presentation: 秋田県総合食品研究センター(秋田)
  • Year and Date: 2011-05-27
• [Presentation] バキュロウイルス感染Sf9昆虫細胞によるヒトインターロイキン-2の生産 (2011)
  • Author(s): 進藤祐宜, 菊地賢一, 後藤猛
  • Organizer: 秋田応用生命科学研究会第18回講演会
  • Place of Presentation: 秋田県総合食品研究センター
  • Year and Date: 2011-05-27
• [Presentation] バキュロウイルス感染Sf9昆虫細胞によるヒトインターロイキン-2生産と糖鎖構造解析 (2011)
  • Author(s): 進藤祐宜, 後藤猛, 菊地賢一
  • Organizer: 化学工学会第76年会
  • Place of Presentation: 東京農工大学(東京)
• [Presentation] バキュロウイルス感染Sf9昆虫細胞によるヒトインターロイキン-2生産と糖鎖構造解析 (2011)
  • Author(s): 進藤祐宜, 後藤猛, 菊地賢一
  • Organizer: 化学工学会第76年会
  • Place of Presentation: 東京農工大学(震災によりWeb開催)
• [Presentation] 昆虫細胞-バキュロウイルス発現におけるアスパルティックプロテアーゼについて (2010)
  • Author(s): 小野洋輝, 後藤猛, 菊地賢一, 韮澤悟, 高橋砂織
  • Organizer: 第76回日本生化学会東北支部例会
  • Place of Presentation: コラッセふくしま(福島)
  • Year and Date: 2010-05-08
• [Presentation] Sf-9昆虫細胞-バキュロウイルス発現系由来アスパルティックプロテアーゼ(SAP)の特性解析 (2010)
  • Author(s): 小野洋輝, 後藤猛, 菊地賢一, 韮澤悟, 高橋砂織
  • Organizer: 秋田化学技術協会講演会
  • Place of Presentation: 秋田大学(秋田)
  • Year and Date: 2010-03-05
• [Presentation] Optimization of soluble human interleukin-2 production in E. coli with coexpression of molecular chaperons in the cold induced system (2010)
  • Author(s): Gotoh, T., Horii, I., Kikuchi, K.-I
  • Organizer: 14th International Biotechnology Symposium and Exhibition
  • Place of Presentation: Palacongressi (Rimini, Italy)
• [Presentation] バキュロウイルス感染Sf-9昆虫細胞培養におけるプロレニンプロセッシング酵素の特性とレニン過剰分解の抑制 (2009)
  • Author(s): 後藤猛, 安和広乃, 日景翔輝, 菊地賢一, 韮澤悟, 高橋砂織
  • Organizer: 第9回食品酵素化学研究会・第15回
  • Place of Presentation: 秋田県総合食品研究センター(秋田)
  • Year and Date: 2009-09-04
• [Presentation] Sf-9昆虫細胞におけるプロレニンの発現とプロレニンプロセッシング酵素の動態 (2009)
  • Author(s): 安和広乃, 後藤猛, 菊地賢一, 韮澤悟, 樋渡一之, 堀一之, 高橋砂織
  • Organizer: 日本生化学会東北支部第75回例会・シンポジウム
  • Place of Presentation: 東北大学(仙台)
  • Year and Date: 2009-05-09
• [Presentation] Expression of recombinant human (rh)-renin in Sf-9 insect cells : Application of rh-renin for inhibitor screening (2009)
  • Author(s): Takahashi, S., Hori, K., Hiwatashi, K., Awa, H., Kikuchi, K.-I., Gotoh, T
  • Organizer: 6th General Meeting of the International Proteolysis Society
  • Place of Presentation: Marriot Hotel (Surfers Paradise, QLD Australia)
• [Presentation] Characterization of prorenin processing enzyme responsible for in situ active human renin production by baculovirus-infected Sf-9 cells and inhibition of excessive processing (2009)
  • Author(s): Gotoh, T., Awa, H., Hikage, S., Kikuchi, K.-I., Takahashi, S
  • Organizer: 14th European Congress on Biotechnology
  • Place of Presentation: Palau de Congressos de Catalunya (Barcelona, Spain)
• [Presentation] Direct production of active human renin by baculovirus-infected Sf-9 insect cell culture (2009)
  • Author(s): Gotoh, T., Awa, H., Kikuchi, K.-I., Takahashi, S
  • Organizer: 8th World Congress of Chemical Engineering
  • Place of Presentation: Palais des Congres de Montreal (Montreal, Canada)
• [Presentation] バキュロウイルス感染Sf-9昆虫細胞培養系におけるキチナーゼの発現 (2009)
  • Author(s): 高橋砂織, 樋渡一之, 菊地賢一, 後藤猛
  • Organizer: 第23回キチンキトサンシンポジウム
  • Place of Presentation: 佐賀大学(佐賀)