プロキラル前駆体からキラルオルガノフォスフォチオ酸の酵素合成
Wen-Shan Li1, Yingchun Li, Craig M Hill
1Department of Chemistry, Texas A&M University, P.O. Box 30012, College Station, Texas 77842-3012, USA.
Journal of the American Chemical Society
|April 4, 2002
まとめ
Pseudomonas diminuta phosphotriesteraseは,特定の酵素基板を選択的に分割する. タンパク質工学は,対照的置換物の水解を可能にし,プロキラル前駆体からキラル産物合成を可能にします.
科学分野:
- バイオカタリシス バイオカタリシス
- 酵素工学とは
- オーガニック・シンセシス オーガニック・シンセシス
背景:
- リン酸トライエステラゼ (PTE) 酵素は,有機リン酸化合物の解毒に不可欠です.
- Pseudomonas diminutaの酵素は,フォスフォートリエステル基板を水解する際にエナチオセレクティブ性を示しています.
- PTEの活性部位を理解し,操作することは,その基板特異性を制御する鍵です.
研究 の 目的:
- ビス-p-ニトロフェニルアルキルフォスフォチオ酸エステルに対する Pseudomonas diminuta phosphotriesterase (PTE) のエナチオ選択性を調査する.
- キラル合成のための変異された基板特異性を持つPTE変種を設計する.
- エンジニアリングされたPTEを使用して,プロキラル前駆体からキラル製品を製造することを実証する.
主な方法:
- ビス-p-ニトロフェニルアルキルフォスフォチオ酸エステルを使用した酵素性水解測定法.
- PTE活性部位のサイト指向型変異.
- チラルクロマトグラフィを用いた製品エナティオメリック過剰の分析.
主要な成果:
- 野生型PTEは,プロ-Rp-ニトロフェノラート置換物を,様々なアルキルフォスフォチオ酸エステルから選択的に割った.
- メチル,エチル,イソプロピル置換剤のエナチオメリック過剰は99%を超えました.
- エンジニアリングされたPTE変種が生成され,プロ-S置換剤を好ましく水解した.
結論:
- Pseudomonas diminuta PTEは,特定の基板水解に対して高いエナチオ選択性を示しています.
- PTE活性部位のタンパク質工学により,エナンチオセレクティビティの逆転が可能です.
- この方法論は,プロキラル基板からキラル化合物の非対称合成のための新しい経路を提供します.
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