Rational design and application of artificial L-amino acid dehydrogenases from sequence database

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K18688
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: University of Shizuoka
• Principal Investigator: NAKANO SHOGO 静岡県立大学, 食品栄養科学部, 助教 (80748541)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: L-スレオニン脱水素酵素 / 祖先型設計 / 完全コンセンサス設計 / 構造機能解析

Outline of Final Research Achievements

Several L-amino acid dehydrogenases are expected to apply to quantify amino acid concentration in plasma. In this study, we utilized L-threonine dehydrogenase as research target. Two artificial SDR-TDHs were designed by utilizing sequence database to improve their enzymatic function. The artificial SDR-TDHs bore high thermal stability, NAD+ affinity and productivity compared with native SDR-TDH. Furthermore, enzyme kinetics parameters of the artificial SDR-TDHs are almost identical to the native SDR-TDH.
In addition, we can elucidate product release mechanism of SDR-TDH through structural and functional analysis of monomeric SDR-TDH.

Free Research Field

タンパク質工学