Exploration and characterization of novel beta-N-acetylglucosaminidase using sub-site recognition inhibitors.

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K05792
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 38020:Applied microbiology-related
• Research Institution: Okayama University
• Principal Investigator: Kanzaki Hiroshi 岡山大学, 環境生命科学学域, 教授 (60183787)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Keywords: β-N-アセチルグルコサミニダーゼ / TMG-chitotriomycin / PNP-TMG / β-GlcNAcase

Outline of Final Research Achievements

We found that insect GlcNAcase is inhibited by a specific inhibitor TMG-chitotriomycin as well as its analog PNP-TMG, but the inhibitory activity of PNP-TMG is about 1000-fold lower than that of TMG-chitotriomycin. On the other hand, GlcNAcase purified from PNP-GlcNAc-assimilating bacterium MK26 was inhibited to the same degree by TMG-chitotriomycin and PNP-TMG, indicating that inhibition specificity of MK26 GlcNAcase against TMG-compounds was significantly different from that of insect GlcNAcase. Although strain MK26 produces GlcNAcases with different molecular weights and other properties depending on the cultivation time, their inhibition specificity against TMG compounds did not alter with cultivation time.

Free Research Field

応用微生物学

Academic Significance and Societal Importance of the Research Achievements

糖質加水分解酵素は，同じファミリーに属していても，そのサブサイト構造の違いにより基質特異性が異なり，得られる生成物が異なることが知られている。その研究は主に，精製酵素の結晶構造解析で進められてきたが，我々が見出したサブサイト認識阻害剤を使うことで，サブサイト構造の異なる酵素の存在を明らかにすることが可能であることが明らかとなった。この阻害剤を用いた研究と，タンパク質立体構造解析研究とを組み合わせることで、より詳細な酵素活性中心の機能解明が可能になると推察される。