Is acidic mammalian chitinase a major glycosidase in the stomachs of mouse and porcine?

• Project/Area Number: 16K07699
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Kogakuin University
• Principal Investigator: Oyama Fumitaka 工学院大学, 先進工学部, 教授 (40194641)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 酸性ほ乳類キチナーゼ / キチン / キチン含有生物 / 糖質分解酵素 / プロテアーゼ耐性 / 昆虫 / 家畜飼料 / カニクイザル / マーモセット / 旧世界ザル / 新世界ザル / 消化酵素 / 胃 / 飼料 / マウス / ブタ / キチナーゼ / N-アセチル-D-グルコサミン (GlcNAc) / 食物繊維 / 酵素 / ペプシン / バイオマス / 糖

Outline of Final Research Achievements

Chitin is a polymer of N-acetyl-D-glucosamine (GlcNAc) and a main constituent of insects’ exoskeleton. Insects are rich in protein with high energy conversion efficiency. We show that mouse, pig, chicken, crab-eating monkey and common marmoset highly express pepsin-, trypsin- and chymotrypsin-resistant AMCase in the stomachs. We also showed that AMCase is most active at pH 2.0-4.0 and degrades chitin and mealworm shells into GlcNAc dimers under gastrointestinal conditions. Thus, AMCase act as digestive enzymes in mouse, pig, chicken, crab-eating monkey and common marmoset.

Academic Significance and Societal Importance of the Research Achievements

AMCase が，マウス，ブタ，ニワトリ，カニクイザル，コモンマーモセット（雑食性）で消化酵素として働き，イヌ（肉食性）とウシ（草食性）ではそのような働きが認められなかったので，食性がその発現レベルに影響し，特定の動物におけるキチンの消化性を決めることを明らかにした。本研究成果は，昆虫などのキチン含有生物を，ほ乳類，鳥類の飼料として丸ごと利用する科学的根拠となり得る。従って，本研究は,「キチン,キチン含有生物を用いた効率的で持続的な家畜生産，世界の食糧供給」に大きく貢献する。

Research Products

• [Int'l Joint Research] Homolka Hospital(チェコ)
• [Int'l Joint Research] Homolka Hospital/Bioinova Ltd.(チェコ)
• [Int'l Joint Research] Mayo Clinic(米国)
• [Journal Article] High expression of acidic chitinase and chitin digestibility in the stomach of common marmoset (Callithrix jacchus)， an insectivorous nonhuman primate (2019)
  • Author(s): Tabata， E.， Kashimura， A.， Uehara， M.， Wakita， S.， Sakaguchi， M.， Sugahara， Y.， Yurimoto， T.， Sasaki， E.， Matoska， V.， Bauer， P. O. & Oyama， F.
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 159-159
  • DOI: 10.1038/s41598-018-36477-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chitinase mRNA Levels Determined by QPCR in Crab-Eating Monkey (Macaca fascicularis) Tissues: Species-Specific Expression of Acidic Mammalian Chitinase and Chitotriosidase (2018)
  • Author(s): Uehara, M., Tabata, E., Ishii, K., Sawa, A., Ohno, M., Sakaguchi, M., Matoska, V., Bauer, P. O. & Oyama, F.
  • Journal Title: Genes (Basel) Volume: 9 Issue: 5 Pages: 244-244
  • DOI: 10.3390/genes9050244
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chitin digestibility is dependent on feeding behaviors, which determine acidic chitinase mRNA levels in mammalian and poultry stomachs (2018)
  • Author(s): Tabata, E., Kashimura, A., Kikuchi, A., Masuda, H., Miyahara, R., Hiruma, Y., Wakita, S., Ohno, M., Sakaguchi, M., Sugahara, Y., Matoska, V., Bauer, P. O. and Oyama, F.
  • Journal Title: Sci. Rep. Volume: 8 Issue: 1 Pages: 1461-1461
  • DOI: 10.1038/s41598-018-19940-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Acidic chitinase-chitin complex is dissociated in a competitive manner by acetic acid: purification of natural enzyme for supplementation purposes (2018)
  • Author(s): Tabata, E., Kashimura, A., Wakita, S., Sakaguchi, M., Sugahara, Y., Imamura, Y., Shimizu, H., Matoska, V., Bauer, P. O. and Oyama, F.
  • Journal Title: Int. J. Mol. Sci. Volume: 19 Issue: 2 Pages: 362-362
  • DOI: 10.3390/ijms19020362
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Gastric and intestinal proteases resistance of chicken acidic chitinase nominates chitin-containing organisms for alternative whole edible diets for poultry. (2017)
  • Author(s): Tabata, E., Kashimura, A., Wakita, S., Ohno, M., Sakaguchi, M., Sugahara, Y., Kino, Y., Matoska, V., Bauer, P.O. and Oyama, F.
  • Journal Title: Sci. Rep. Volume: 7 Issue: 1 Pages: 6662-6662
  • DOI: 10.1038/s41598-017-07146-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Protease resistance of porcine acidic mammalian chitinase under gastrointestinal conditions implies that chitin-containing organisms can be sustainable dietary resources (2017)
  • Author(s): Tabata, E., Kashimura, A., Wakita, S., Ohno, M., Sakaguchi, M., Sugahara, Y., Imamura, Y., Seki, S., Ueda, H., Matoska, V., Bauer, P. O. and Oyama, F.
  • Journal Title: Sci. Rep. Volume: 7 Issue: 1 Pages: 12963-12963
  • DOI: 10.1038/s41598-017-13526-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Improved fluorescent labeling of chitin oligomers: chitinolytic properties of acidic mammalian chitinase under somatic tissue pH conditions. (2017)
  • Author(s): Wakita, S., Kimura, M., Kato, N., Kashimura, A., Kobayashi, S., Kanayama, N., Ohno, M., Honda, S., Sakaguchi, M., Sugahara, Y., Bauer, P.O. and Oyama, F.
  • Journal Title: Carbohydr. Polym. Volume: 164 Pages: 145-153
  • DOI: 10.1016/j.carbpol.2017.01.095
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mouse acidic mammalian chitinase exhibits transglycosylation activity at somatic tissue pH. (2017)
  • Author(s): Wakita S, Kobayashi S, Kimura M, Kashimura A, Honda S, Sakaguchi M, Sugahara Y, Kamaya M, Matoska V, Bauer PO, and Oyama F.
  • Journal Title: FEBS Lett. Volume: 591 Issue: 20 Pages: 3310-3318
  • DOI: 10.1002/1873-3468.12798
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Acidic mammalian chitinase is a proteases-resistant glycosidase in mouse digestive system. (2016)
  • Author(s): Ohno, M.; Kimura, M.; Miyazaki, H.; Okawa, K.; Onuki, R.; Nemoto, C.; Tabata, E.; Wakita, S.; Kashimura, A.; Sakaguchi, M.; Sugahara, Y.; Nukina, N.; Bauer, P. O.; Oyama, F.
  • Journal Title: Sci. Rep. Volume: 6 Issue: 1 Pages: 37756-37756
  • DOI: 10.1038/srep37756
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Loss and gain of human acidic mammalian chitinase activity by nonsynonymous SNPs. (2016)
  • Author(s): Okawa, K.; Ohno, M.; Kashimura, A.; Kimura, M.; Kobayashi, Y.; Sakaguchi, M.; Sugahara, Y.; Kamaya, M.; Kino, Y.; Bauer, P. O.; Oyama, F.
  • Journal Title: Mol. Biol. Evol. Volume: 33 Issue: 12 Pages: 3183-3193
  • DOI: 10.1093/molbev/msw198
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Functional properties of mouse chitotriosidase expressed in the periplasmic space of Escherichia coli. (2016)
  • Author(s): Kimura, M.; Wakita, S.; Ishikawa, K.; Sekine, K.; Yoshikawa, S.; Sato, A.; Okawa, K.; Kashimura, A.; Sakaguchi, M.; Sugahara, Y.; Yamanaka, D.; Ohno, N.; Bauer, P. O.; Oyama, F.
  • Journal Title: PLoS One Volume: 11 Issue: 10 Pages: e0164367-e0164367
  • DOI: 10.1371/journal.pone.0164367
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] ブタ酸性キチナーゼは，消化器系プロテアーゼに耐性で，キチン分解酵素として機能する (2019)
  • Author(s): 小山 文隆、樫村 昭徳、脇田 悟誌、菅原 康里、田畑 絵理
  • Organizer: 日本農芸化学会2019年度(H31年度)大会
• [Presentation] Acidic mammalian chitinase is a major chitinase in stomach resistant to pepsin and trypsin/chymotrypsin digestion (2018)
  • Author(s): Oyama, F., Kimura, M., Okawa, K., Onuki, R., Nemoto, C., Tabata, E., Wakita, S., Kashimura, A., Sakaguchi, M., Sugahara, Y., Ohno, M.
  • Organizer: 14th International Chitin and Chitosan Conference (14th ICCC)
  • Int'l Joint Research
• [Presentation] Gene expression analysis of chitinases in crab-eating monkey: species-specific expression of acidic mammalian chitinase and chitotriosidase (2018)
  • Author(s): Uehara, M., Tabata, E., Ohno, M., Sakaguchi, M., Oyama, F.
  • Organizer: 14th International Chitin and Chitosan Conference (14th ICCC)
  • Int'l Joint Research
• [Presentation] Acidic chitinase mRNA levels and their chitinolytic activity are affected by the feeding behavior, which may determine chitin digestibility in animals (2018)
  • Author(s): Tabata, E., Kashimura, A., Wakita, S., Ohno, M., Sakaguchi, M., Sugahara, Y., Oyama, F.
  • Organizer: 14th International Chitin and Chitosan Conference (14th ICCC)
  • Int'l Joint Research
• [Presentation] Quantification of chitinases mRNA levels by qPCR in crab-eating monkey tissues: Comparison with mouse and human (2018)
  • Author(s): Uehara, M., Tabata, E., Ohno, M., Sakaguchi, M., Oyama, F.
  • Organizer: The 67th Annual Meeting of the American Society of Human Genetics
  • Int'l Joint Research
• [Presentation] Feeding behaviors determine acidic chitinase mRNA levels in mammalian and poultry stomachs. (2018)
  • Author(s): Oyama, F., Kashimura, A., Kikuchi, A., Masuda, H., Miyahara, R., Hiruma, Y., Wakita, S., Ohno, M., Sakaguchi, M., Sugahara, Y., Tabata, E.
  • Organizer: The 67th Annual Meeting of the American Society of Human Genetics
  • Int'l Joint Research
• [Presentation] Gene expression analysis of mammalian chitinases in common marmoset (Callithrix jacchus) tissue. (2018)
  • Author(s): Tabata, E., Kashimura, A., Uehara, M. Wakita, S., Sakaguchi, M., Sugahara, Y., Yurimoto, T., Sasaki, E., Oyama, F.
  • Organizer: The 67th Annual Meeting of the American Society of Human Genetics
  • Int'l Joint Research
• [Presentation] ニワトリ酸性キチナーゼの消化酵素としての機能解析 (2018)
  • Author(s): 田畑絵理，樫村昭德，脇田悟誌，大野美紗，坂口政吉，菅原康里，小山文隆
  • Organizer: 日本農芸化学会2018年度大会[名古屋]
• [Presentation] ヒト酸性ほ乳類キチナーゼの nonsynonymous SNPs による活性の調節 (2018)
  • Author(s): 小山文隆，大野美紗，樫村昭徳，木村将大，坂口政吉，菅原康里，大川一明
  • Organizer: 日本農芸化学会2018年度大会[名古屋]
• [Presentation] 2種類のマウスほ乳類キチナーゼのキチナーゼ活性比較 (2018)
  • Author(s): 木村将大，梅山隆敏，脇田悟誌，大川一明，坂口政吉，小山文隆
  • Organizer: 日本農芸化学会2018年度大会[名古屋]
• [Presentation] ニワトリ組織におけるキチナーゼ遺伝子発現レベルとその酵素学的機能の解析 (2017)
  • Author(s): 田畑絵理，樫村昭德，大野美紗，坂口政吉，小山文隆
  • Organizer: 日本農芸化学会2017年度大会
  • Place of Presentation: 京都市
  • Year and Date: 2017-03-18
• [Presentation] Gene expression analysis and enzyme function of pig mammalian chitinase (2017)
  • Author(s): Tabata, E., Kashimura, A., Wakita, S., Ohno, M., Sakaguchi, M., Sugahara, Y., Oyama, F.
  • Organizer: The 67th Annual Meeting of the American Society of Human Genetics (ASHG) (Orlando)
  • Int'l Joint Research
• [Presentation] Mechanistic insight of inactivation of mouse chitinase-like protein Ym1 (2017)
  • Author(s): Oyama, F., Sakaguchi, M., Sugahara, Y., Okawa, K.
  • Organizer: The 67th Annual Meeting of the American Society of Human Genetics (ASHG) (Orlando)
  • Int'l Joint Research
• [Presentation] Chitinases mRNA levels quantified by qPCR in crab-eating monkey tissues. (2017)
  • Author(s): Kimura, M., Umeyama, T., Wakita, S., Okawa, K., Sakaguchi, M., Oyama, F.
  • Organizer: The 67th Annual Meeting of the American Society of Human Genetics (ASHG) (Orlando)
  • Int'l Joint Research
• [Presentation] ニワトリ酸性キチナーゼの遺伝子発現解析とその酵素機能 (2017)
  • Author(s): 田畑絵理，樫村昭徳，脇田悟志，大野美紗，菅原康里，坂口政吉，小山文隆
  • Organizer: 第31回日本キチン・キトサン学会大会
• [Presentation] ヒト酸性ほ乳類キチナーゼの進化学的解析 (2017)
  • Author(s): 小山文隆，大野美紗，樫村昭徳，木村将大，坂口政吉，菅原康里，大川一明
  • Organizer: 第31回日本キチン・キトサン学会大会
• [Presentation] カニクイザルにおけるキチナーゼ遺伝子の発現解析 (2017)
  • Author(s): 上原麻衣子，田畑絵里，大野美紗，坂口政吉，小山文隆
  • Organizer: 第31回日本キチン・キトサン学会大会
• [Presentation] マウスほ乳類キチナーゼの比較解析 (2017)
  • Author(s): 木村将大, 梅山隆敏, 脇田悟誌, 大川一明, 坂口政吉, 小山文隆
  • Organizer: 第31回日本キチン・キトサン学会大会
• [Presentation] キチンオリゴ糖蛍光標識法を用いた酸性ほ乳類キチナーゼの特性解析 (2017)
  • Author(s): 脇田悟誌，木村将大, 樫村明徳, 坂口政吉, 菅原康里, 小山文隆
  • Organizer: 第31回日本キチン・キトサン学会大会
• [Presentation] ヒト酸性ほ乳類キチナーゼの活性喪失とその活性化 (2017)
  • Author(s): 小山文隆
  • Organizer: 第6回 医薬工3大学 包括連携推進シンポジウム 特別講演（東京薬科大学）
  • Invited
• [Presentation] Gene expression analysis of mammalian chitinase transcripts in pig tissues. (2016)
  • Author(s): Tabata, E., Ohno, M., Sakaguchi, M., Oyama, F.
  • Organizer: The 66th Annual Meeting of the American Society of Human Genetics (ASHG)
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2016-10-20
  • Int'l Joint Research
• [Presentation] Inactivation of mouse AMCase by introducing amino acid substitutions encoded by single-nucleotide polymorphisms in human AMCase gene. (2016)
  • Author(s): Oyama, F., Okawa, K., Ohno, M., Kashimura, A. Kobayashi, Y. Sakaguchi, M., Sugahara, Y.
  • Organizer: The 66th Annual Meeting of the American Society of Human Genetics (ASHG)
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2016-10-19
  • Int'l Joint Research
• [Presentation] マウス胃における酸性ほ乳類キチナーゼの発現と生産細胞の同定 (2016)
  • Author(s): 小山文隆，大野美紗，木村将大，大川一明，田畑絵理，脇田悟誌，樫村昭徳， 坂口政吉，菅原康里
  • Organizer: 第30回 日本キチン・キトサン学会大会
  • Place of Presentation: 川越市
  • Year and Date: 2016-08-19
• [Presentation] ブタ組織におけるキチナーゼ遺伝子発現レベルの解析 (2016)
  • Author(s): 田畑絵理，大野美紗，坂口政吉，小山文隆
  • Organizer: 第30回 日本キチン・キトサン学会大会
  • Place of Presentation: 川越市
  • Year and Date: 2016-08-18