Studies on glycosylation of chitinase secreted by marine bacteria

• Project/Area Number: 19780163
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Fisheries chemistry
• Research Institution: Nihon University
• Principal Investigator: ITOI Shiro Nihon University, 生物資源科学部, 講師 (30385992)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,760,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥360,000); Fiscal Year 2008: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2007: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: キチナーゼ / 海洋細菌 / 糖鎖修飾 / 結晶キチン / Vibrio proteolyticus / α型結晶キチン

Research Abstract

海洋細菌Vibrio proteolyticus由来キチナーゼの分子量が明らかに理論値よりも大きいことから、翻訳後修飾の可能性が高いと考え、研究を進めた。当該タンパク質の大腸菌組換え体が複数のレクチンを用いるレクチンブロッティングにより陽性反応を示した。また、850アミノ酸からなる当該組換えキチナーゼの一次構造のうち、C末端側250アミノ酸を欠損する組換え体を作製し、レクチンブロッティングを試みた結果、C末端側欠損組換え体でも陽性反応が認められ、この領域中に糖鎖修飾部位が存在する可能性が示唆された。

Research Products

• [Journal Article] Phenotypic variation in Lactococcus lactis subsp. lactis isolates derived from intestinal tracts of marine and freshwater fishes (2009)
  • Author(s): Shiro Itoi Kohei Yuasa, Sayka Washio. Takeshi Abe, Erika Ikuno and Haruo Sugita
  • Journal Title: Journal of Applied Microbiology (印刷中)
  • Peer Reviewed
• [Journal Article] Phenotypic variation in Lactococcus lactis subsp. lactis isolates derived from intestinal tracts of marine and freshwater fishes (2009)
  • Author(s): Shiro Itoi, Kohei Yuasa, Sayaka Washio, Takeshi Abe, Erika Ikuno and Haruo Sugita
  • Journal Title: J. Appl. Microbiol (印刷中)
  • Peer Reviewed
• [Journal Article] Purification, characterization and cloning of Vibrio parahaemolyticus chitinolytic enzymes and application to oligosaccharide production (2008)
  • Author(s): Kazunari Kadokura, Yusuke Sakamoto, Akiko Rokutani, Takanori Ikegami, Takao Hirano, Mahiro Yamamoto, Kaori Saito, Wataru Hakamata, Shiro Itoi, Haruo Sugita, Tadatake Oku and Toshiyuki Nishio
  • Journal Title: Journal of Applied Glycoscience 55 Pages: 157-164
  • NAID: 10020738847
  • Peer Reviewed
• [Journal Article] Purification, characterization and cloning of Vibrio parahaemolyticus chitinolytic enzymes and application to oligosaccharide production (2008)
  • Author(s): Kazunari Kadokura, Yusuke Sakamoto, Akiko Rokutani, Takanori Ikegami, Takao Hirano, Mahiro Yamamoto, Kaori Saito, Wataru Hakamata, Shiro Itoi, Haruo Sugita, Tadatake Oku and Toshivuki Nishio
  • Journal Title: J. Appl. Glycosci 2 Pages: 157-164
  • NAID: 10020738847
  • Peer Reviewed
• [Journal Article] Identification of a novel endochitinase from a marine bacterium Vibrio proteolyticus strain No. 442 (2007)
  • Author(s): Shiro Itoi, Yuna Kanomata, Yuki Koyama, Kazunari Kadokura, Shinsuke Uchida, Toshiyuki Nishio, Tadatake Oku and Haruo Sugita
  • Journal Title: Biochimica et Biophysica Acta 1774 Pages: 1099-1107
  • Peer Reviewed
• [Journal Article] Purification and characterization of Vibrio parahaemolyticus extracellular chitinase and chitin oligosaccharide deacetylase involved in the production of heterodisaccharide from chitin (2007)
  • Author(s): Kazunari Kadokura, Akiko Rokutani, Masahiro Yamamoto, Takanori Ikegami, Haruo Sugita, Shiro Itoi, Wataru Hakamata, Tadatake Oku and Toshiyuki Nishio
  • Journal Title: Applied Microbiology and Biotechnology 75 Pages: 357-365
  • Peer Reviewed
• [Journal Article] Identification of a novel endochitinase from a marine bacterium Vibrio proteolyticus strain No. 442. (2007)
  • Author(s): Shiro Itoi, Yuna Kanomata, Yuki Koyama, Kazunari Kadokura, Shinsuke Uchida, Toshiyuki Nishio, Tadatake Oku and Haruo Sugita
  • Journal Title: Biochim. Biophys. Acta 1774 Pages: 1099-1107
  • Peer Reviewed
• [Journal Article] Purification and characterization of Vibrio parahaemolyticus extracellular chitinase and chitin oligosaccharide deacetylase involved in the production of heterodisaccharide from chitin. (2007)
  • Author(s): Kazunari Kadokura, Akiko Rokutani, Masahiro Yamamoto, Takanori Ikegami, Haruo Sugita, Shiro Itoi, Wataru Hakamata, Tadatake Oku and Toshiyuki Nishio
  • Journal Title: Appl. Microbiol. Biotechnol. 75 Pages: 357-365
  • Peer Reviewed
• [Presentation] 海洋細菌Vibrio proteolyticus由来キチナーゼのpH依存性に及ぼすC末端領域の影響 (2007)
  • Author(s): 鹿股悠奈、糸井史朗、門倉一成、西尾俊幸、奥忠武、杉田治男
  • Organizer: 平成19年度日本水産学会秋季大会
  • Place of Presentation: 函館(北海道大学水産学部)
  • Year and Date: 2007-09-26
• [Presentation] 海洋細菌Vibrio proteolyticus由来キチナーゼのpH依存性に及ぼすC末端領域の影響 (2007)
  • Author(s): 鹿股悠奈、糸井史朗、門倉一成、西尾俊幸、奥忠武、杉田治男
  • Organizer: 平成19年度日本水産学会秋季大会
  • Place of Presentation: 北海道大学(函館)
  • Year and Date: 2007-09-26
• [Presentation] 海洋細菌Vibrio proteolyticus由来エンドキチナーゼの精製および活性におよぼすC末端領域の影響 (2007)
  • Author(s): 鹿股悠奈、糸井史朗、内田晋輔、門倉一成、西尾俊幸、奥忠武、杉田治男
  • Organizer: 2007(平成19)年度日本水産増殖学会大会
  • Place of Presentation: 品川(東京海洋大学)
  • Year and Date: 2007-07-18
• [Presentation] 海洋細菌Vibrio proteolyticus由来エンドキチナーゼの精製および活性におよぼすC末端領域の影響 (2007)
  • Author(s): 鹿股悠奈、糸井史朗、内田晋輔、門倉一成、西尾俊幸、奥忠武、杉田治男
  • Organizer: 2007(平成19)年度日本水産増殖学会大会
  • Place of Presentation: 東京海洋大学(品川)
  • Year and Date: 2007-07-18