Development of stealth-nanoparticles avoiding animal immune system that mimics the cell surface of filamentous fungi.

• Project/Area Number: 15K07348
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Tohoku University
• Principal Investigator: Yoshimi Akira 東北大学, 未来科学技術共同研究センター, 准教授 (60436102)
• Co-Investigator(Renkei-kenkyūsha): ABE Keietsu 東北大学, 農学研究科, 教授 (50312624) ; KAWAKAMI Kazuyoshi 東北大学, 医学系研究科, 教授 (10253973) ; TAKAMI Seiichi 東北大学, 多元物質科学研究所, 准教授 (40311550)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 糸状菌 / 細胞壁 / ステルス素材 / α-1,3-グルカン / オリゴ糖合成 / 免疫細胞応答試験 / マウス免疫試験 / ステルス新素材 / オリゴ糖酵素合成 / 免疫応答評価 / オリゴ糖 / 転移酵素合成 / マウス試験 / 医療用ナノ粒子 / 亜臨界水分解

Outline of Final Research Achievements

This study aims to create medical nanoparticles with stealth function using α - 1,3 - glucan (AG) and hydrophobin (HP) which are novel stealth materials derived from filamentous fungi. The results obtained in this study are as follows.
1) Determination of purification conditions for AG and AG oligosaccharides. 2) Confirmation of the stealth function of AG and AG oligosaccharides. 3) Confirmation of non-interaction between AG and HP. 4) Determination of enzymatic synthesis conditions of AG and AG oligosaccharides. 5) Demonstration of the stealth function of AG in vivo. 6) Evaluation of adsorption conditions of AG oligosaccharides and the particles.7) Discovery of galactosaminogalactan (GAG) as hyphal adhesion factor in Aspergillus oryzae. 8) Demonstration of additive interactions on the stealth function of AG and GAG.

Research Products

• [Journal Article] Function and biosynthesis of cell wall α-1,3-glucan in fungi (2017)
  • Author(s): Yoshimi A., Miyazawa K., Abe K.*
  • Journal Title: J. Fungi Volume: 3 Issue: 4 Pages: 63-63
  • DOI: 10.3390/jof3040063
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of Cell Wall α-1,3-Glucan–Deficient Mutants in <i>Aspergillus oryzae</i> Isolated by a Screening Method Based on Their Sensitivities to Congo Red or Lysing Enzymes (2017)
  • Author(s): Yoshimi A., Hirama M., Tsubota Y., Kawakami K., Zhang S., Gomi K., and K. Abe*
  • Journal Title: Journal of Applied Glycoscience Volume: 64 Issue: 3 Pages: 65-73
  • DOI: 10.5458/jag.jag.JAG-2017_004
  • NAID: 130005988426
  • ISSN: 1344-7882, 1880-7291
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of cell wall α-1,3-glucan-deficient mutants in Aspergillus oryzae which were isolated by a screening method based on their sensitivities to Congo red or Lysing Enzymes. (2017)
  • Author(s): Akira Yoshimi, Misa Hirama, Yasunobu Tsubota, Kazuyoshi Kawakami, Silai Zhang, Katsuya Gomi, and Keietsu Abe
  • Journal Title: Journal of Applied Glycolscience Volume: submitted
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cell wall a-1,3-glucan prevents a-amylase adsorption onto fungal cell in submerged culture of Aspergillus oryzae. (2017)
  • Author(s): Zhang S., Sato H., Ichinose S., Tanaka M., Miyazawa K., Yoshimi A., Abe K., Shintani T., K. Gomi
  • Journal Title: J. Biosci. Bioengi. Volume: 124 Issue: 1 Pages: 47-53
  • DOI: 10.1016/j.jbiosc.2017.02.013
  • Peer Reviewed / Open Access
• [Journal Article] Cell wall structure and biogenesis in Aspergillus species (2016)
  • Author(s): Akira Yoshimi, Ken Miyazawa, Keietsu Abe
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: published online Issue: 9 Pages: 1-12
  • DOI: 10.1080/09168451.2016.1177446
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Substantial decrease in cell wall α-1,3-glucan caused by disruption of the kexB gene encoding a subtilisin-like processing protease in Aspergillus oryzae (2016)
  • Author(s): Osamu Mizutani, Matsuko Shiina, Akira Yoshimi, Motoaki Sano, Takeshi Watanabe, Youhei Yamagata, Tasuku Nakajima, Katsuya Gomi, and Keietsu Abe
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: published online Issue: 9 Pages: 1-11
  • DOI: 10.1080/09168451.2016.1158632
  • Peer Reviewed
• [Presentation] 産業用糸状菌Aspergillus oryzaeのGPI型糖転移酵素AgtAの生物学的機能解析 (2018)
  • Author(s): 小泉亜未, 矢野成和, 宮澤拳, 吉見啓, 佐野元昭, 阿部敬悦
  • Organizer: 日本農芸化学会2018年度大会
• [Presentation] Asp30 and Asp73 of Aspergillus oryzae cutinase CutL1 are involved in the ionic interaction with fungal hydrophobin RolA (2018)
  • Author(s): Yuki Terauchi, Yoon-Kyung Kim, Takumi Tanaka, Kei Nanatani, Akira Yoshimi, Toru Takahashi, Keietsu Abe
  • Organizer: 14th European Conference on Fungal Genetics (ECFG14)
  • Int'l Joint Research
• [Presentation] Comparative analysis of the function of α-1,3-glucan syntheses, AgsA and AgsB, in Aspergillus nidulans. (2017)
  • Author(s): Ken Miyazawa, Akira Yoshimi, Shigekazu Yano, Shin Kasahara, Humihiko Hasegawa, Keietsu Abe
  • Organizer: 29th Fungal Genetic Conference
  • Place of Presentation: Pacific Grove, California, USA
  • Year and Date: 2017-03-14
  • Int'l Joint Research / Invited
• [Presentation] Comparative analysis of the function of α-1,3-glucan syntheses, AgsA and AgsB, in Aspergillus nidulans. (2017)
  • Author(s): Ken Miyazawa, Akira Yoshimi, Shigekazu Yano, Shin Kasahara, Humihiko Hasegawa, Keietsu Abe
  • Organizer: 14th International Aspergillus meeting
  • Place of Presentation: Pacific Grove, California, USA
  • Year and Date: 2017-03-13
  • Int'l Joint Research
• [Presentation] 麹菌界面活性タンパク質hydrophobin RolA とCutinase CutL1 間の相互作用に関与するCutL1側新奇相互作用部位D30 (2017)
  • Author(s): 寺内祐貴，金允卿，田中拓未，七谷圭，吉見啓，高橋徹，阿部敬悦
  • Organizer: 生物工学会2017年度北日本支部 福島シンポジウム
• [Presentation] Aspergillus nidulansの２種のα-1,3-グルカン合成酵素による産生多糖の化学構造解析 (2017)
  • Author(s): 宮澤拳，吉見啓，矢野成和，笠原紳，阿部敬悦
  • Organizer: 生物工学会2017年度北日本支部 福島シンポジウム
• [Presentation] 麹菌Aspergillus oryzaeの液体振盪培養における菌糸接着機構の解析 (2017)
  • Author(s): 宮澤拳，吉見啓，古明地敬介，田畑風華，佐野元昭，阿部敬悦
  • Organizer: 第17回糸状菌分子生物学コンファレンス
• [Presentation] 麹菌Aspergillus oryzaeの液体培養において菌糸接着に関与する多糖の特定とその欠損株の表現型解析 (2017)
  • Author(s): 宮澤拳, 吉見啓, 田畑風華, 古明地敬介, 佐野元昭, 阿部敬悦
  • Organizer: 日本応用糖質科学会平成２９年度大会（第６６回）
• [Presentation] Aspergillus属菌における菌糸凝集因子の解析と高密度培養による物質高生産への応用 (2017)
  • Author(s): 吉見啓、宮澤拳 、佐野元昭、古明地敬介 、張斯来、五味勝也、阿部敬悦
  • Organizer: 日本菌学会第61回大会（第2回環境微生物系学会合同大会2017）
• [Presentation] Asp30 of Aspergillus oryzae cutinase CutL1 is involved in the ionic interaction with fungal hydrophobin RolA (2017)
  • Author(s): Yuki Terauchi, Yoon-Kyung Kim, Takumi Tanaka, Kei Nanatani, Akira Yoshimi, Toru Takahashi, and Keietsu Abe
  • Organizer: IFBC2017 (International Fungal Biology Conference)
  • Int'l Joint Research
• [Presentation] The adsorption kinetics and the self-assembled structures of Aspergillus oryzae hydrophobin RolA on various chemically modified solid surfaces (2017)
  • Author(s): Yuki Terauchi, Megumi Nagayama, Takumi Tanaka, Hiroki Tanabe, Toshihiko Arita, Hiroshi Yabu, Takeshi Higuchi, Kei Nanatani, Akira Yoshimi, and Keietsu Abe
  • Organizer: IFBC2017 (International Fungal Biology Conference)
  • Int'l Joint Research
• [Presentation] Comparative analysis of the function of α-1,3-glucan synthases, AgsA and AgsB, in model filamentous fungus Aspergillus nidulans (2017)
  • Author(s): Ken Miyazawa, Akira Yoshimi, Shin Kasahara, Shigekazu Yano, and Keietsu Abe
  • Organizer: IFBC2017 (International Fungal Biology Conference)
  • Int'l Joint Research
• [Presentation] Aspergillus oryzaeのα-1,3-グルカンおよびガラクトサミノガラクタン欠損株における菌糸完全分散性 (2017)
  • Author(s): 吉見啓, 宮澤拳, 佐野元昭, 古明地敬介, 張斯来, 五味勝也, 阿部敬悦
  • Organizer: 日本応用糖質科学会東北支部会講演会
• [Presentation] Aspergillus nidulansにおける二種のα-1,3-グルカン合成酵素の機能の比較解析 (2017)
  • Author(s): 宮澤拳, 吉見啓, 笠原紳, 矢野成和, 阿部敬悦
  • Organizer: 日本応用糖質科学会東北支部会講演会
• [Presentation] Aspergillus nidulansのAgsA, AgsBが合成するα-1,3-グルカンの化学構造の比較解析 (2016)
  • Author(s): 宮澤拳，吉見啓，阿部敬悦
  • Organizer: 第16回糸状菌分子生物学コンファレンス
  • Place of Presentation: 京都大学宇治キャンパス、京都府・宇治市
  • Year and Date: 2016-11-16
• [Presentation] コンゴレッド感受性に基づく麹菌の細胞壁多糖α-1,3-グルカン低減株の分離とその性質 (2016)
  • Author(s): 吉見啓、平間美紗、坪田康信、川上和義、阿部敬悦
  • Organizer: 日本応用糖質科学会平成２８年度大会（第６５回）
  • Place of Presentation: 福山大学宮地茂記念館、広島県・福山市
  • Year and Date: 2016-09-14
• [Presentation] Growth characteristics and protein productivity of Aspergillus oryzae quintuple disruption mutants of α-1,3-glucan-related genes (2016)
  • Author(s): Ken Miyazawa, Akira Yoshimi, Silai Zhang, Motoaki Sano, Katsuya Gomi, Fumihiko Hasegawa, Keietsu Abe
  • Organizer: 13th European Conference on Fungal Genetics (ECFG13)
  • Place of Presentation: Paris, France
  • Year and Date: 2016-04-03
  • Int'l Joint Research
• [Presentation] 糸状菌の細胞壁ストレス応答シグナル伝達系と細胞壁α-1,3-グルカンの生物学的機能 (2016)
  • Author(s): 阿部敬悦、吉見啓
  • Organizer: 応用糖質科学会 東日本支部シンポジウム
  • Place of Presentation: 東京大学農学部 弥生講堂一条ホール、東京都・文京区
  • Invited
• [Presentation] 宮澤拳、吉見啓、張斯来、佐野元昭、笠原紳、五味勝也、阿部敬悦 (2016)
  • Author(s): Aspergillus 属菌間における細胞壁α-1,3-グルカン生合成機構の比較解析
  • Organizer: 日本応用糖質科学会東北支部会講演会
  • Place of Presentation: 岩手大学農学部、岩手県・盛岡市
• [Presentation] Aspergillus 属菌における細胞壁α-1,3-グルカン研究からの新展開 (2016)
  • Author(s): 吉見啓
  • Organizer: 糸状菌遺伝子研究会第３７回例会
  • Place of Presentation: 北とぴあ、東京都・北区
  • Invited
• [Presentation] 細胞壁多糖α-1,3-グルカンが低減した麹菌をスクリーニングする方法の開発 (2015)
  • Author(s): 吉見 啓、平間美沙、坪田康信、川上和義、阿部敬悦
  • Organizer: 第15回糸状菌分子生物学コンファレンス
  • Place of Presentation: ルミエール府中、東京都
  • Year and Date: 2015-11-19
• [Presentation] 麹菌におけるα-1,3-グルカン生合成関連遺伝子群の５重破壊株における培養性状と物質生産性の評価 (2015)
  • Author(s): 宮澤拳、吉見 啓、張斯来、佐野元昭、五味勝也、阿部敬悦
  • Organizer: 第15回糸状菌分子生物学コンファレンス
  • Place of Presentation: ルミエール府中、東京都
  • Year and Date: 2015-11-19
• [Presentation] 糸状菌Aspergillus nidulansにおける細胞壁構築シグナル伝達経路の解析と細胞壁多糖の分析 (2015)
  • Author(s): 吉見 啓、宮澤拳、阿部敬悦
  • Organizer: 日本応用糖質科学会平成27年度大会（第64回）
  • Place of Presentation: 奈良春日野国際フォーラム 甍～I・RA・KA～、奈良市
  • Year and Date: 2015-09-16
• [Presentation] 麹菌におけるα-1,3-グルカン合成酵素遺伝子とアミラーゼ遺伝子の５重破壊株における培養性状 (2015)
  • Author(s): 宮澤拳、吉見 啓、張斯来、佐野元昭、五味勝也、阿部敬悦
  • Organizer: 第7回 日本応用糖質科学会東北支部会講演会
  • Place of Presentation: 弘前大学、弘前市
  • Year and Date: 2015-07-11
• [Book] 酵母菌・麹菌・乳酸菌の産業応用展開、第９章麹菌の細胞壁α-1,3グルカン欠損株による高密度培養と物質高生産への利用 (2018)
  • Author(s): 吉見啓、宮澤拳、張斯来
  • Total Pages: 264
  • Publisher: シーエムシー出版
• [Book] 化学と教育誌、ヘッドライン：カビ ～世間の嫌われ者、実は身近で役立つ善玉カビ～ (2017)
  • Author(s): 吉見啓，宮澤拳，阿部敬悦
  • Total Pages: 47
  • Publisher: 日本化学会
• [Book] 化学と教育誌、ヘッドライン「カビー世間の嫌われ者、実は身近で役立つ善玉カビー」 (2017)
  • Author(s): 吉見 啓、宮澤 拳、阿部 敬悦
  • Publisher: 日本化学会
• [Patent(Industrial Property Rights)] 変異型糸状菌及び当該変異型糸状菌を用いた物質生産方法 (2017)
  • Inventor(s): 阿部敬悦，吉見啓、宮澤拳、田畑風華、五味勝也、佐野元昭
  • Industrial Property Rights Holder: 阿部敬悦，吉見啓、宮澤拳、田畑風華、五味勝也、佐野元昭
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2017-091734
  • Filing Date: 2017
• [Patent(Industrial Property Rights)] 麹菌の細胞壁変異株の選抜方法、当該細胞壁変異株、当該細胞壁変異株を用いた米製品およびその製造方法 (2015)
  • Inventor(s): 吉見啓，阿部敬悦，川上和義，平間美沙，坪田康信
  • Industrial Property Rights Holder: 吉見啓，阿部敬悦，川上和義，平間美沙，坪田康信
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-205934
  • Filing Date: 2015-10-19