Search, characterization, and application of new chitinase from marine organisms

• Project/Area Number: 25450309
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Aquatic life science
• Research Institution: Nihon University
• Principal Investigator: MATSUMIYA Masahiro 日本大学, 生物資源科学部, 教授 (60150702)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: キチナーゼ / 精製 / 機能解析 / ｃＤＮＡクローニング / 発現 / 魚類 / 軟体動物 / 甲殻類 / 遺伝子 / キチン / cDNAクローニング / 胃

Outline of Final Research Achievements

Chitinase is a useful enzyme for enzymatic production of chitin oligosaccharides which are bioactive substance. Therefore, we conducted purification, characterization, molecular cloning, and expression of chitinases from several kinds of marine organisms.　Six kinds of chitinases were purified and characterized from two species of fish stomach, Parapristipoma trilineatum and Sardinops melanostictus, and two species of octopus salivary gland, Octopus vulgaris and Octopus dofleini. Eleven kinds of full length chitinase cDNA were obtained from four species of fish stomach, two species of octopus salivary gland, one species of cuttlefish liver, and one species of sea hare ovotestis. Expression vector system of chitinases from Sebastiscus marmoratus stomach and shark stomach were constructed and expressions were conducted using E. coli and yeast, respectively.

Research Products

• [Journal Article] Stomach chitinase from Japanese sardine Sardinops melanostictus: purification, characterization, and molecular cloning of chitinase isozymes with a long linker (2016)
  • Author(s): Satoshi Kawashima, Hiroki Ikehata, Chihiro Tada, Tomohiro Ogino, Hiromi Kakizaki, Mana Ikeda, Hideto Fukushima and Masahiro Matsumiya
  • Journal Title: Marine Drugs Volume: 14 Issue: 1 Pages: 1-13
  • DOI: 10.3390/md14010022
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Molecular cloning of a chitinase gene from the ovotestis of Kuroda’s sea hare Aplysia kurodai (2016)
  • Author(s): ３）Gaku Matsunaga, Syuuji Karasuda, Ryo Nishino, Hideto Fukushima, Masahiro Matsumiya
  • Journal Title: Advances in Bioscience and Biotechnology Volume: 7 Issue: 01 Pages: 38-46
  • DOI: 10.4236/abb.2016.71005
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Distribution of chitinolytic enzymes in the organs and cDNA cloning of chitinase isozymes from the stomach of two species of fish, chub mackerel (Scomber japonicus) and silver croaker (Pennahia argentata) (2015)
  • Author(s): Hiromi Kakizaki, Mana Ikeda, Hideto Fukushima, and Masahiro Matsumiya
  • Journal Title: Open Journal of Marine Science Volume: ５ Issue: 04 Pages: 398-411
  • DOI: 10.4236/ojms.2015.54032
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] 第１回キチン・キトサンフォーラムに参加して (2015)
  • Author(s): 松宮政弘
  • Journal Title: キチン・キトサン研究 Volume: 21 Pages: 53-54
• [Journal Article] Primary structure and enzymatic properties of chitinase isozymes purified from the stomach of the marbled rockfish Sebastiscus marmoratus (2014)
  • Author(s): Mana Ikeda, Daisuke Shirase, Takuya Sato, Mika Ueda, Shinichi Hirabayashi, and Masahiro Matsumiya
  • Journal Title: Journal of Chitin and Chitosan Science Volume: 2(2) Issue: 2 Pages: 106-116
  • DOI: 10.1166/jcc.2014.1048
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cloning of a novel chitinase cDNA from the stomach of the coelacanth Latimeria chalumnae (Sarcopterygii) (2014)
  • Author(s): Hiromi Kakizaki, Kaneyuki Hamaguchi, Mana Ikeda, and Masahiro Matsumiya
  • Journal Title: Journal of Chitin and Chitosan Science Volume: 2(2) Issue: 2 Pages: 123-129
  • DOI: 10.1166/jcc.2014.1051
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Purification of a chitinase from the posterior salivary gland of common octopus Octopus vulgaris and its properties (2014)
  • Author(s): Tomohiro Ogino, Hirotaka Tabata, Mana Ikeda, Hiromi Kakizaki, and Masahiro Matsumiya
  • Journal Title: Journal of Chitin and Chitosan Science Volume: 2(2) Issue: 2 Pages: 135-142
  • DOI: 10.1166/jcc.2014.1049
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Molecular cloning of a novel chitinase gene from blue shark (Prionace glauca; Chondrichthyes) stomach (2014)
  • Author(s): Teruhisa Suzuki, Hiromi Kakizaki, Mana Ikeda, and Masahiro Matsumiya
  • Journal Title: Journal of Chitin and Chitosan Science Volume: 2(2) Issue: 2 Pages: 143-148
  • DOI: 10.1166/jcc.2014.1050
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Purification and characterization of a liver chitinase from golden cuttlefish, Sepia esculenta (2014)
  • Author(s): Ryo Nishino, Akiyoshi Suyama, Mana Ikeda, Hiromi Kakizaki, and Masahiro Matsumiya
  • Journal Title: Journal of Chitin and Chitosan Science Volume: 2(3) Issue: 3 Pages: 238-243
  • DOI: 10.1166/jcc.2014.1065
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Molecular cloning of multiple chitinase genes in swimming crab Portunus trituberculatus (2014)
  • Author(s): Naoya Fujitani, Hirotaka Hasegawa, Hiromi Kakizaki, Mana Ikeda, and Masahiro Matsumiya
  • Journal Title: Journal of Chitin and Chitosan Science Volume: 2(2) Issue: 2 Pages: 149-156
  • DOI: 10.1166/jcc.2014.1046
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Purification, characterization, and molecular cloning of chitinases from the stomach of threeline grunt Parapristipoma trilineatum (2013)
  • Author(s): Mana Ikeda, Yujiro Kondo, Masahiro Matsumiya
  • Journal Title: Process Biochem. Volume: 48 Issue: 9 Pages: 1324-1334
  • DOI: 10.1016/j.procbio.2013.06.016
  • Peer Reviewed
• [Presentation] 海洋生物のキチン分解酵素 (2015)
  • Author(s): 松宮政弘
  • Organizer: 日本応用糖質科学会・東日本支部シンポジウム
  • Place of Presentation: 藤沢商工会館（神奈川県・藤沢市）
  • Year and Date: 2015-11-20
  • Invited
• [Presentation] ホシザメ胃由来キチナーゼ遺伝子のクローニングおよび異種宿主発現 (2015)
  • Author(s): 川島聖史・水野寛己・柿崎博美・池田愛・福島英登・上田光宏・松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 東北大学（宮城県・仙台市）
  • Year and Date: 2015-09-23
• [Presentation] アメフラシ Aplysia kurodai生殖腺キチナーゼのcDNAクローニング (2015)
  • Author(s): 松永岳・西野亮・柿崎博美・福島英登･松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 東北大学（宮城県・仙台市）
  • Year and Date: 2015-09-23
• [Presentation] アオリイカにおけるキチナーゼの体内分布とcDNAクローニング (2015)
  • Author(s): 西野亮・柿崎博美・池田愛・福島英登・松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 東北大学（宮城県・仙台市）
  • Year and Date: 2015-09-23
• [Presentation] マダコ唾液腺、盲嚢および中腸腺に存在するキチナーゼの性状比較と中腸腺キチナーゼのcDNAクローニング (2015)
  • Author(s): 荻野智大・柿崎博美・池田愛・福島英登・松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 東北大学（宮城県・仙台市）
  • Year and Date: 2015-09-23
• [Presentation] Molecular cloning of chitinase gene from the kidney of five fish species (2015)
  • Author(s): Hiromi Kakizaki, Satoshi Kawashima, Taro Tsukamoto, Miku Fujiwara, Mana Ikeda, Hideto Fukushima, and Masahiro Matsumiya
  • Organizer: 13th International Conference on Chitin and Chitosan
  • Place of Presentation: Munster, Germany
  • Year and Date: 2015-08-30
  • Int'l Joint Research
• [Presentation] Distribution of chitinase and cellulase activity in the several organs of a fish, largescale blackfish Girella punctate (2015)
  • Author(s): Kazumasa Kajiwara, Ryo Nishino, Hideto Fukushima, Masahiro Matsumiya
  • Organizer: 13th International Conference on Chitin and Chitosan
  • Place of Presentation: Munster, Germany
  • Year and Date: 2015-08-30
  • Int'l Joint Research
• [Presentation] Molecular cloning of a chitinase from the liver of Japanese common squid Todarodes pacificus (2015)
  • Author(s): Ryo Nishino, Hiromi Kakizaki, Mana Ikeda, Hideto Fukushima, Masahiro Matsumiya
  • Organizer: 13th International Conference on Chitin and Chitosan
  • Place of Presentation: Munster, Germany
  • Year and Date: 2015-08-30
  • Int'l Joint Research
• [Presentation] Molecular cloning of a chitinase gene from the stomach of cloudy catshark Scyliorhinus torazame (Chondrichthyes) (2015)
  • Author(s): Satoshi Kawashima, Hiromi Kakizaki, Kaneyuki Hamaguchi, Mana Ikeda, Hideto Fukushima, Masahiro Matsumiya
  • Organizer: 13th International Conference on Chitin and Chitosan
  • Place of Presentation: Munster, Germany
  • Year and Date: 2015-08-30
  • Int'l Joint Research
• [Presentation] ハイギョにおけるキチン分解酵素の体内分布と消化管キチナーゼの性状 (2015)
  • Author(s): 柿崎博美，上野 翼， 池田 愛， 福島英登， 松宮政弘
  • 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（熊本県・熊本市）
  • Year and Date: 2015-08-21
• [Presentation] 数種硬骨魚類におけるキチン分解酵素の体内分布とcDNAクローニング (2015)
  • Author(s): 柿崎博美, 池田 愛, 藤原美玖, 福島英登, 松宮政弘
  • 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（熊本県・熊本市）
  • Year and Date: 2015-08-21
• [Presentation] カサゴ胃由来キチナーゼの大腸菌による発現 (2015)
  • Author(s): 松永 岳、柿崎博美、池田 愛、福島英登、上田光宏、松宮政弘
  • 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（熊本県・熊本市）
  • Year and Date: 2015-08-20
• [Presentation] マダコ盲嚢キチナーゼの精製と性状 (2015)
  • Author(s): 荻野智大, 柿崎博美, 池田 愛, 福島英登, 松宮政弘
  • 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（熊本県・熊本市）
  • Year and Date: 2015-08-20
• [Presentation] Glycoside hydrolase family18 chitinase from the stomach of fish: characteristics of isozymes (2015)
  • Author(s): Masahiro Matsumiya, Hiromi Kakizaki, and Mana Ikeda
  • Organizer: 29th Annual Symposium of the Protein Society
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2015-07-22
  • Int'l Joint Research
• [Presentation] マイワシ胃キチナーゼアイソザイムの性状と一次構造の比較 (2014)
  • Author(s): 川島聖史・池端弘樹・多田千紘・柿崎博美・池田愛・上田光宏・松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 九州大学
  • Year and Date: 2014-09-20
• [Presentation] 数種魚類におけるキチン分解酵素の体内分布とキチナーゼのcDNAクローニング (2014)
  • Author(s): 柿崎博美・岩田龍一・勝瀬竜土・塚本太郎・藤原美玖・池田愛・松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 九州大学
  • Year and Date: 2014-09-20
• [Presentation] 軟骨魚類ホシザメおよびヨシキリザメ胃におけるキチナーゼの存在と性質 (2014)
  • Author(s): 鈴木輝久・今村良・柿崎博美・池田愛・松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 九州大学
  • Year and Date: 2014-09-20
• [Presentation] ２種のタコにおけるキチン分解酵素の体内分布とミズダコ唾液腺キチナーゼの性状 (2014)
  • Author(s): 荻野智大・柿崎博美・池田愛・松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 九州大学
  • Year and Date: 2014-09-20
• [Presentation] ガザミ中腸腺キチナーゼの性状とcDNAクローニング (2014)
  • Author(s): 藤谷直矢・長谷川浩隆・柿崎博美・池田愛・松宮政弘
  • Organizer: 日本水産学会秋季大会
  • Place of Presentation: 九州大学
  • Year and Date: 2014-09-20
• [Presentation] ２種のタコ中腸腺キチナーゼの性状 (2014)
  • Author(s): 荻野智大・柿崎博美・池田 愛・松宮政弘
  • Organizer: 第２8回キチン・キトサンシンポジウム
  • Place of Presentation: 順天堂大学
  • Year and Date: 2014-08-07
• [Presentation] Structure and Function of Chitinase Isozymes from the Stomach of the Silver Croaker Pennahia argentatus
  • Author(s): Mana Ikeda and Masahiro Matsumiya
  • Organizer: 11th International Conference of the European Chitin society
  • Place of Presentation: Porto, Portugal
• [Presentation] Molecular cloning of a novel chitinase gene from the stomach of the blue shark Prionace glauca (Chondrichthyes)
  • Author(s): T. Suzuki, H. Kakizaki, M. Ikeda, M. Matsumiya
  • Organizer: 10th Asia-Pacific Chitin Chitosan Symposium
  • Place of Presentation: Yonago, Japan
• [Presentation] Molecular cloning of multiple chitinase genes in swimming crab Portunus trituberculatus
  • Author(s): N. Fujitani, H. Hasegawa, H. Kakizaki, M. Ikeda, M. Matsumiya
  • Organizer: 10th Asia-Pacific Chitin Chitosan Symposium
  • Place of Presentation: Yonago, Japan
• [Presentation] Primary structure and enzymatic properties of chitinase isozymes purified from the stomach of the Marbled rockfish Sebastiscus marmoratus
  • Author(s): M. Ikeda, D. Shirase, T. Sato, M. Ueda, S. Hirabayashi, M. Matsumiya
  • Organizer: 10th Asia-Pacific Chitin Chitosan Symposium
  • Place of Presentation: Yonago, Japan
• [Presentation] cDNA cloning of a novel chitinase from the stomach of Coelacanth Latimeria chalumnae (Sarcopterygii)
  • Author(s): H. Kakizaki, K. Hamaguchi, M. Ikeda, M. Matsumiya
  • Organizer: 10th Asia-Pacific Chitin Chitosan Symposium
  • Place of Presentation: Yonago, Japan
• [Presentation] Purification and characterization of a chitinase from the salivary gland of the common octopus Octopus vulgaris
  • Author(s): T. Ogino, H. Tabata, M. Ikeda, M. Matsumiya
  • Organizer: 10th Asia-Pacific Chitin Chitosan Symposium
  • Place of Presentation: Yonago, Japan
• [Presentation] Purification, characterisaiton, and cDNA cloning of a chitinase from the liver of Golden cuttlefish Sepia esculenta
  • Author(s): R.Nishino, A. Suyama, M. Ikeda, H. Kakizaki, M. Matsumiya
  • Organizer: 10th Asia-Pacific Chitin Chitosan Symposium
  • Place of Presentation: Yonago, Japan
• [Presentation] Fish stomach chitinases: Presence of the isozyme groups and enzymatic properties of those
  • Author(s): Masahiro Matsumiya, Mana Ikeda
  • Organizer: 10th Asia-Pacific Chitin Chitosan Symposium
  • Place of Presentation: Yonago, Japan
  • Invited
• [Presentation] ミズダコ中腸腺キチナーゼの精製と性質
  • Author(s): 荻野智大・柿崎博美・池田愛・松宮政弘
  • Organizer: 日本水産学会春季大会
  • Place of Presentation: 北海道大学