Characterization of a recombinant glucosidase from Ensifer adhaerens NBRC 100388 and evaluation of its glucosyl transfer activity
| Project/Area Number |
17K07739
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Oyama National College of Technology |
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Principal Investigator |
UEDA MAKOTO 小山工業高等専門学校, 物質工学科, 教授 (10615751)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2019: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2018: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
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| Keywords | 配糖化 / テルペンアルコール / エンシファー / グルコシダーゼ / リナロール / 配糖体 / 微生物反応 / 応用微生物 / 酵素 / 発酵 / バイオテクノロジー / 糖 |
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| Outline of Final Research Achievements |
An α-glucosidase gene from a strain of Ensifer adhaerens NBRC 100388 showing transglucosylation activity toward alkylalcohols was cloned and expressed in Escherichia coli. The molecular mass of the purified recombinant α-glucosidase protein was estimated to be 60 kDa. This α-glucosidase from E. adhaerens NBRC 100388 (EaG) was able to hydrolyze maltose, maltotriose, maltotetraose, maltulose, sucrose, trehalose, ethyl-α-D-glucoside, and p-nitrophenyl1;D-glucoside (pNPG). The enzyme activity increased in the presence of NH4+ and K+ ions but was inhibited by Cu+ and Zn2+. The enzyme exhibited optimal activity with maltose at a pH of 7.5 and a temperature of 45 °C. The transglucosylation activity was observed not only for nerol, a primary alcohol, but also for 6-gingerol, a secondary alcohol, and (-)-linalool, a tertiary alcohol. The enantioselectivity for the glucosyl transfer towards the acceptor of (±)-linalool was 21.5% ee for the (-)-isomer.
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| Academic Significance and Societal Importance of the Research Achievements |
アグリコンとなるステロイドやテルペンアルコールは、立体構造により薬理作用や香りなどの生物活性が異なる。また、配糖体が生物活性を持つ場合もあり、α-アルブチンはチロシナーゼの阻害活性を持ち、ポリフェノール配糖体は糖の結合位置で抗酸化能が変わる（文献3）。さらに、配糖体のα-とβ-アノマーは、分解酵素の存在など生物的な環境により安定性も異なる。以上のことから、アグリコンに対し立体・位置選択性を示し、アノマーの作り分けも可能な酵素による配糖体の精密合成が可能になれば、様々なアグリコンの配糖体や、また同じアグリコンから機能の異なる配糖体を、新たな機能性素材として創出できる。
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Report
(4 results)
Research Products
(7 results)
